Noncanonical functions of the serine-arginine-rich splicing factor (SR) family of proteins in development and disease.

Members of the serine/arginine (SR)-rich protein family of splicing factors play versatile roles in RNA processing steps and are often essential for normal development. Dynamic changes in RNA processing and turnover allow fast cellular adaptions to a changing microenvironment and thereby closely cooperate with transcription factor networks that establish cell identity within tissues. SR proteins play fundamental roles in the processing of pre-mRNAs by regulating constitutive and alternative splicing. More recently, SR proteins have also been implicated in other aspects of RNA metabolism such as mRNA stability, transport and translation. The- emerging noncanonical functions highlight the multifaceted functions of these SR proteins and identify them as important coordinators of gene expression programmes. Accordingly, most SR proteins are essential for normal cell function and their misregulation contributes to human diseases such as cancer.

Self-generated strategies in the phonological similarity effect.

Strategy use is an important source of individual differences during immediate serial reconstruction. However, not all strategies are equally suited for all tasks. Therefore, assessing participants' dynamic strategy selection across contexts is an important next step for reliable interpretation of individual differences in short-term memory span - in both experimental and clinical settings. Strategy use during reconstruction of phonologically similar and phonologically distinct word sets was directly assessed using a self-report questionnaire. In two experiments, participants reported consistent use of phonological strategies across word sets; however, participants reported additionally using non-phonological strategies (i.e., mental imagery and sentence generation) when tasked with remembering phonologically similar words. In particular, strategy selection was most impacted when the phonologically similar word set was either the only word set or the first word set participants received. When the phonologically similar lists were presented after a classic list of phonologically distinct words, participants continued using the phonological strategies that had been effective for the distinct lists. Moreover, in both experiments, accuracy of phonologically similar lists was better predicted by use of non-phonological strategies than use of phonological strategies. Specifically, reported use of verbalization or rehearsal did not predict accuracy, but participants who reported regularly using mental imagery and/or sentence generation (typically in conjunction with rehearsal) displayed greater serial memory for similar words. These results do not undermine the general assumptions of the phonological similarity effect, but they do indicate that its interpretation is less straightforward than previously thought.

The Detection of Foreign Items in Laundry Industry by Dual-Energy X-ray Transmission-Advantages and Limits.

Firefighters, paramedics, nursing staff, and other occupational groups are in constant need of fast and proper cleaning of their professional workwear, not only during a pandemic. Thus, laundry technology needs to become more efficient and automated. Unfortunately, some steps of the cleaning process, such as finding and removing foreign items from pockets or belts, are still completed manually. This is not just time-consuming but potentially dangerous for the workers due to the hazardous nature of items such as scissors, scalpels, or syringes. Additionally, some items may damage the garments by staining or harm the laundry machines, causing malfunctions and process failure. On the one hand, these foreign items are often hidden inside the clothes, making detection very challenging with conventional superficial sensors. On the other hand, these items can be diverse and cannot be detected by metal detectors alone. X-ray transmission has proven to be a powerful tool for detecting items inside of objects. The dual-energy approach (DE-XRT) even allows obtaining quantitative information about the chemical composition of the measured materials. In this study, working garments were accompanied and filled with realistic foreign items. The potential of DE-XRT to detect those items was successfully shown.

Verification of a National Emission Inventory and Influence of On-road Vehicle Manufacturer-Level Emissions.

Road vehicles make important contributions to a wide range of pollutant emissions from the street level to global scales. The quantification of emissions from road vehicles is, however, highly challenging given the number of individual sources involved and the myriad factors that influence emissions such as fuel type, emission standard, and driving behavior. In this work, we use highly detailed and comprehensive vehicle emission remote sensing measurements made under real driving conditions to develop new bottom-up inventories that can be compared to official national inventory totals. We find that the total UK passenger car and light-duty van emissions of nitrogen oxides (NOx) are underestimated by 24-32%, and up to 47% in urban areas, compared with the UK national inventory, despite agreement within 1.5% for total fuel used. Emissions of NOx at a country level are also shown to vary considerably depending on the mix of vehicle manufacturers in the fleet. Adopting the on-road mix of vehicle manufacturers for six European countries results in up to a 13.4% range in total emissions of NOx. Accounting for the manufacturer-specific fleets at a country level could have a significant impact on emission estimates of NOx and other pollutants across the European countries, which are not currently reflected in emission inventories.

Potential of Dual and Multi Energy XRT and CT Analyses on Iron Formations.

Dual and multi energy X-ray transmission imaging (DE-/ME-XRT) are powerful tools to acquire quantitative material characteristics of diverse samples without destruction. As those X-ray imaging techniques are based on the projection onto the imaging plane, only two-dimensional data can be obtained. To acquire three-dimensional information and a complete examination on topology and spatial trends of materials, computed tomography (CT) can be used. In combination, these methods may offer a robust non-destructive testing technique for research and industrial applications. For example, the iron ore mining and processing industry requires the ratio of economic iron minerals to siliceous waste material for resource and reserve estimations, and for efficient sorting prior to beneficiation, to avoid equipment destruction due to highly abrasive quartz. While XRT provides information concerning the thickness, areal density and mass fraction of iron and the respective background material, CT may deliver size, distribution and orientation of internal structures. Our study shows that the data provided by XRT and CT is reliable and, together with data processing, can be successfully applied for distinguishing iron oxide rich parts from waste. Furthermore, heavy element bearing minerals such as baryte, uraninite, galena and monazite can be detected.

Underestimated Ammonia Emissions from Road Vehicles.

In this study, we use comprehensive vehicle emission remote sensing measurements of over 230,000 passenger cars to estimate total UK ammonia (NH3) emissions. Estimates are made using "top-down" and "bottom-up" methods that demonstrate good agreement to within 1.1% for total fuel consumed or CO2 emitted. A central component of this study is the comprehensive nature of the bottom-up emission estimates that combine highly detailed remote sensing emission data with over 4000 km of 1 Hz real driving data. Total annual UK NH3 emissions from gasoline passenger cars are estimated to be 7.8 ± 0.3 kt from the bottom-up estimate compared with 3.0 ± 1.7 kt reported by the UK national inventory. An important conclusion from the analysis is that both methodologies confirm that gasoline passenger car NH3 emissions are underestimated by a factor of about 2.6 compared with the 2018 UK National Atmospheric Emissions Inventory. Furthermore, we find that inventory estimates of urban emissions of NH3 for passenger cars are underestimated by a factor of 17.

Energy depletion and not ROS formation is a crucial step of glucolipotoxicity (GLTx) in pancreatic beta cells.

We have shown previously that genetic or pharmacological deletion of KATP channels protect against beta cell dysfunction induced by reactive oxygen species (ROS). Since it is assumed that glucolipotoxicity (GLTx) causes ROS production, we aimed to evaluate whether suppression of KATP channel activity can also prevent beta cell damage evoked by GLTx. We used an in vitro model of GLTx and measured distinct parameters of stimulus-secretion coupling. GLTx gradually induced disturbances of Ca2+ oscillations over 3 days. This impairment in Ca2+ dynamics was partially reversed in beta cells without functional KATP channels (SUR1-/-) and by the sulfonylurea gliclazide but not by tolbutamide. By contrast, the GLTx-induced suppression of glucose-induced insulin secretion could not be rescued by decreased KATP channel activity pointing to a direct interaction of GLTx with the secretory capacity. Accordingly, GLTx also suppressed KCl-induced insulin secretion. GLTx was not accompanied by decisively increased ROS production or enhanced apoptosis. Insulin content of beta cells was markedly reduced by GLTx, an effect not prevented by gliclazide. Since GLTx markedly diminished the mitochondrial membrane potential and cellular ATP content, lack of ATP is assumed to decrease insulin biosynthesis. The deleterious effect of GLTx is therefore caused by direct interference with the secretory capacity whereby reduction of insulin content is one important parameter. These findings deepen our understanding how GLTx damages beta cells and reveal that GLTx is disconnected from ROS formation, a notion important for targeting beta cells in the treatment of diabetes. Overall, GLTx-induced energy depletion may be a primary step in the cascade of events leading to loss of beta cell function in type-2 diabetes mellitus.

Body dissatisfaction and suicidal ideation among psychiatric inpatients with eating disorders.

The current study the relationship between eating disorders (EDs) and suicidal ideation and suicide attempt in adult inpatients. In particular, the present study investigated one potential mechanism, body dissatisfaction (BD), which may contribute to increased risk for suicide in adult ED patients. A sample of 432 psychiatric inpatients ranging from 18 to 65 years of age participated in the current study. Findings indicated that patients who have higher levels of BD also had higher levels of passive and active suicidal ideation and previous suicide attempts. Higher levels of BD were also related to increased suicidal ideation after controlling for depression and emotion dysregulation. Although additional risk factors for suicide should be investigated in adults with EDs, this study provides evidence regarding the relationship between BD and risk for suicide ideation and attempt.

Expression of Hsp27 and Hsp70 and vacuolization in the pituitary glands in cases of fatal hypothermia.

Hypothermia causes systemic cellular stress. The pituitary gland is an endocrine gland and plays an important role in thermoregulation. When the core body temperature drops, the pituitary gland is activated by stimulation of hypothalamic hormones. In this study, we investigated morphological alterations of the pituitary gland in cases of fatal hypothermia. Several morphological alterations of the anterior lobe of the pituitary gland, such as hemorrhage, vacuolization, and hyperemia, have been previously described in fatal hypothermia. However, the diagnostic value of these findings is controversial. We compared 11 cases of fatal hypothermia with 10 cases lacking antemortem hypothermic influences. In the presence of thermal cellular stress, the expression of heat shock proteins increases to protect cellular structures. Therefore, we immunohistochemically analyzed Hsp27 and Hsp70. Hsp27 expression was detected in 27.3% of the cases of fatal hypothermia and in 10.0% of the control cases, whereas Hsp70 expression was not detected in any case. Additionally, Sudan staining was performed to quantify fatty degeneration. A positive reaction was found in 45.5% of the study group and in 10.0% of the control group. This indicates that fatty degeneration might be a valuable marker when other macroscopic signs of hypothermia are absent.

[Matching bite marks using modeling wax plates].

Bite marks are rarely seen in forensic medical examinations. In most cases one can easily differentiate whether a bite mark is of human or animal origin due to the arched shape and crushed character of human bite marks. From a forensic point of view it is more interesting whether a dental imprint has been caused by an adult or a child, because children are often abused for a defensive statement. Two cases of bite injuries in children are presented. In the first case, a two-month-old infant was presented by pediatricians. The second case deals with a one-year-old boy who had allegedly been bitten several times by a two-year-old girl in a nursery school. A comparison could be performed between the bite mark impressions on the victim's skin (based on photographic documentation) and dental imprints of single teeth measuring the tooth width on the one hand and the intercanine distance on the other hand with the help of modeling wax plates. This procedure showed good correlations between the bite marks on the victim and the dental status of the biter. Especially for taking bite impressions on children, modeling wax plates are a simple, fast and cheap alternative to dental impression-taking.

Targeted ablation of the Pde6h gene in mice reveals cross-species differences in cone and rod phototransduction protein isoform inventory.

Phosphodiesterase-6 (PDE6) is a multisubunit enzyme that plays a key role in the visual transduction cascade in rod and cone photoreceptors. Each type of photoreceptor utilizes discrete catalytic and inhibitory PDE6 subunits to fulfill its physiological tasks, i.e. the degradation of cyclic guanosine-3',5'-monophosphate at specifically tuned rates and kinetics. Recently, the human PDE6H gene was identified as a novel locus for autosomal recessive (incomplete) color blindness. However, the three different classes of cones were not affected to the same extent. Short wave cone function was more preserved than middle and long wave cone function indicating that some basic regulation of the PDE6 multisubunit enzyme was maintained albeit by a unknown mechanism. To study normal and disease-related functions of cone Pde6h in vivo, we generated Pde6h knock-out (Pde6h(-/-)) mice. Expression of PDE6H in murine eyes was restricted to both outer segments and synaptic terminals of short and long/middle cone photoreceptors, whereas Pde6h(-/-) retinae remained PDE6H-negative. Combined in vivo assessment of retinal morphology with histomorphological analyses revealed a normal overall integrity of the retinal organization and an unaltered distribution of the different cone photoreceptor subtypes upon Pde6h ablation. In contrast to human patients, our electroretinographic examinations of Pde6h(-/-) mice suggest no defects in cone/rod-driven retinal signaling and therefore preserved visual functions. To this end, we were able to demonstrate the presence of rod PDE6G in cones indicating functional substitution of PDE6. The disparities between human and murine phenotypes caused by mutant Pde6h/PDE6H suggest species-to-species differences in the vulnerability of biochemical and neurosensory pathways of the visual signal transduction system.

Does being overweight or male increase a patient's risk of not being referred for an eating disorder consult?

OBJECTIVE: To examine whether sex, age, body mass index (BMI), and eating disorder diagnosis were associated with referral rates for eating disorder consults in a general inpatient psychiatric facility. METHOD: An inpatient sample of 136 individuals with a current eating disorder diagnosis were utilized for this study. Chi square and logistic regression were used to identify variables affecting the likelihood of having an eating disorder consult. RESULTS: More women than men were identified as having a current eating disorder diagnosis. For both women and men, eating disorder not otherwise specified (EDNOS) was the most common diagnosis, followed by binge-eating disorder (BED). Compared to women, there were more men who did not receive an eating disorder consult. Specifically, overweight men were less likely to be referred for a consult. DISCUSSION: Results highlight the need of healthcare providers to ask all individuals about their eating habits, which could then provide an opportunity to explore eating disorder symptoms. Due to historical biases, the less distinct nature of some diagnoses or the absence of symptoms that have been considered indicators of clinical severity, providers may be more likely to overlook individuals with diagnoses of EDNOS and BED for consults, especially men. © 2016 Wiley Periodicals, Inc. (Int J Eat Disord 2016; 49:963-966).

The Bacillus subtilis tyrZ gene encodes a highly selective tyrosyl-tRNA synthetase and is regulated by a MarR regulator and T box riboswitch.

UNLABELLED: Misincorporation of D-tyrosine (D-Tyr) into cellular proteins due to mischarging of tRNA(Tyr) with D-Tyr by tyrosyl-tRNA synthetase inhibits growth and biofilm formation of Bacillus subtilis. Furthermore, many B. subtilis strains lack a functional gene encoding D-aminoacyl-tRNA deacylase, which prevents misincorporation of D-Tyr in most organisms. B. subtilis has two genes that encode tyrosyl-tRNA synthetase: tyrS is expressed under normal growth conditions, and tyrZ is known to be expressed only when tyrS is inactivated by mutation. We hypothesized that tyrZ encodes an alternate tyrosyl-tRNA synthetase, expression of which allows the cell to grow when D-Tyr is present. We show that TyrZ is more selective for L-Tyr over D-Tyr than is TyrS; however, TyrZ is less efficient overall. We also show that expression of tyrZ is required for growth and biofilm formation in the presence of D-Tyr. Both tyrS and tyrZ are preceded by a T box riboswitch, but tyrZ is found in an operon with ywaE, which is predicted to encode a MarR family transcriptional regulator. Expression of tyrZ is repressed by YwaE and also is regulated at the level of transcription attenuation by the T box riboswitch. We conclude that expression of tyrZ may allow growth when excess D-Tyr is present. IMPORTANCE: Accurate protein synthesis requires correct aminoacylation of each tRNA with the cognate amino acid and discrimination against related compounds. Bacillus subtilis produces D-Tyr, an analog of L-Tyr that is toxic when incorporated into protein, during stationary phase. Most organisms utilize a D-aminoacyl-tRNA deacylase to prevent misincorporation of D-Tyr. This work demonstrates that the increased selectivity of the TyrZ form of tyrosyl-tRNA synthetase may provide a mechanism by which B. subtilis prevents misincorporation of D-Tyr in the absence of a functional D-aminoacyl-tRNA deacylase gene.

Chromogranin A is a T cell antigen in human type 1 diabetes.

Chromogranin A (ChgA) is a beta cell secretory granule protein and a peptide of ChgA, WE14, was recently identified as a ligand for diabetogenic CD4 T cell clones derived from the NOD mouse. In this study we compared responses of human CD4 T cells from recent onset type 1 diabetic (T1D) and control subjects to WE14 and to an enzymatically modified version of this peptide. T cell responders to antigens were detected in PBMCs from study subjects by an indirect CD4 ELISPOT assay for IFN-γ. T1D patients (n = 27) were recent onset patients within one year of diagnosis, typed for HLA-DQ8. Controls (n = 31) were either 1st degree relatives with no antibodies or from the HLA-matched general population cohort of DAISY/TEDDY. A second cohort of patients (n = 11) and control subjects (n = 11) was tested at lower peptide concentrations. We found that WE14 is recognized by T cells from diabetic subjects vs. controls in a dose dependent manner. Treatment of WE14 with transglutaminase increased reactivity to the peptide in some patients. This work suggests that ChgA is an important target antigen in human T1D subjects and that post-translational modification may play a role in its reactivity and relationship to disease.

Children's rehearsal development parallels that of self-talk in other executive functions.

Rehearsal is a form of self-talk used to support short-term memory. Historically, the study of rehearsal development has diverged from the study of self-talk more generally. The current experiment examines whether two characteristics of self-talk (impact of task difficulty and self-talk's narrative vs. planning purpose) are also observed in rehearsal. Eighty children, ages 4-7, were tasked with remembering the three-item and six-item lists over a 15-s delay. Children's spontaneous use of fixed rehearsal (i.e., immediate repetition of a just-presented item) and cumulative rehearsal (i.e., cycling through multiple items at a time) was documented from video recordings. Four-year-olds narrated item presentations using fixed rehearsal. Six- and seven-year-olds proactively planned for recall by engaging cumulative rehearsal. Five-year-olds used both forms of rehearsal, but their frequency of cumulative rehearsal was dependent on list length. Therefore, rehearsal is susceptible to task manipulations that affect other forms of self-talk. (PsycInfo Database Record (c) 2024 APA, all rights reserved).

The dynamic genetic determinants of increased transcriptional divergence in spermatids.

Cis-genetic effects are key determinants of transcriptional divergence in discrete tissues and cell types. However, how cis- and trans-effects act across continuous trajectories of cellular differentiation in vivo is poorly understood. Here, we quantify allele-specific expression during spermatogenic differentiation at single-cell resolution in an F1 hybrid mouse system, allowing for the comprehensive characterisation of cis- and trans-genetic effects, including their dynamics across cellular differentiation. Collectively, almost half of the genes subject to genetic regulation show evidence for dynamic cis-effects that vary during differentiation. Our system also allows us to robustly identify dynamic trans-effects, which are less pervasive than cis-effects. In aggregate, genetic effects were strongest in round spermatids, which parallels their increased transcriptional divergence we identified between species. Our approach provides a comprehensive quantification of the variability of genetic effects in vivo, and demonstrates a widely applicable strategy to dissect the impact of regulatory variants on gene regulation in dynamic systems.

Human type 1 diabetes is associated with T cell autoimmunity to zinc transporter 8.

Recently we demonstrated that zinc transporter 8 (ZnT8) is a major target of autoantibodies in human type 1 diabetes (T1D). Because the molecules recognized by T1D autoantibodies are typically also targets of autoreactive T cells, we reasoned that this would likely be the case for ZnT8. To test this hypothesis, IFN-γ-producing T cells specific for ZnT8 in the peripheral blood of 35 patients with T1D (<6 mo after onset at blood draw) and 41 age-matched controls were assayed by ELISPOT using a library of 23 overlapping dipeptide pools covering the entire 369 aa primary sequence. Consistent with our hypothesis, patients showed significantly higher T cell reactivity than the matched controls, manifest in terms of the breadth of the overall response and the magnitude of responses to individual pools. Therefore, the median number of pools giving positive responses (stimulation index ≥ 3) in the control group was 1.0 (range, 0-7) compared with 6.0 (range, 1-20; p < 0.0001) for the patients. Similarly, the median stimulation index of positive responses in controls was 3.1 versus 5.0 in the patients (p < 0.0001). Individually, 7 of 23 pools showed significant disease association (p < 0.001), with several of the component peptides binding the disease associated HLA-DR3 (0301) and -DR4 (0401) molecules in vitro. We conclude that ZnT8 is also a major target of disease-associated autoreactive T cells in human T1D, and we suggest that reagents that target ZnT8-specific T cells could have therapeutic potential in preventing or arresting the progression of this disease.

Emission ratio determination from road vehicles using a range of remote emission sensing techniques.

The development of remote emission sensing techniques such as plume chasing and point sampling has progressed significantly and is providing new insight into vehicle emissions behaviour. However, the analysis of remote emission sensing data can be highly challenging and there is currently no standardised method available. In this study we present a single data processing approach to quantify vehicle exhaust emissions measured using a range of remote emission sensing techniques. The method uses rolling regression calculated over short time intervals to derive the characteristics of diluting plumes. We apply the method to high time-resolution plume chasing and point sampling data to quantify gaseous exhaust emission ratios from individual vehicles. Data from a series of vehicle emission characterisation experiments conducted under controlled conditions is used to demonstrate the potential of this approach. First, the method is validated through comparison with on-board emission measurements. Second, the ability of this approach to detect changes in NOx / CO2 ratios associated with aftertreatment system tampering and different engine operating conditions is shown. Third, the flexibility of the approach is demonstrated by varying the pollutants used as regression variables and quantifying the NO2 / NOx ratios for different vehicle types. A higher proportion of total NOx is emitted as NO2 when the selective catalytic reduction system of the measured heavy duty truck is tampered. In addition, the applicability of this approach to urban environments is illustrated using mobile measurements conducted in Milan, Italy in 2021. Emissions from local combustion sources are distinguished from a complex urban background and the spatiotemporal variability in emissions is shown. The mean NOx / CO2 ratio of 1.61 ppb/ppm is considered representative of the local vehicle fleet. It is envisaged that this approach can be used to quantify emissions from a range of mobile and stationary fuel combustion sources, including non-road vehicles, ships, trains, boilers and incinerators.

Zonadhesin is essential for species specificity of sperm adhesion to the egg zona pellucida.

Interaction of rapidly evolving molecules imparts species specificity to sperm-egg recognition in marine invertebrates, but it is unclear whether comparable interactions occur during fertilization in any vertebrate species. In mammals, the sperm acrosomal protein zonadhesin is a rapidly evolving molecule with species-specific binding activity for the egg zona pellucida (ZP). Here we show using null mice produced by targeted disruption of Zan that zonadhesin confers species specificity to sperm-ZP adhesion. Sperm capacitation selectively exposed a partial von Willebrand D domain of mouse zonadhesin on the surface of living, motile cells. Antibodies to the exposed domain inhibited adhesion of wild-type spermatozoa to the mouse ZP but did not inhibit adhesion of spermatozoa lacking zonadhesin. Zan(-/-) males were fertile, and their spermatozoa readily fertilized mouse eggs in vitro. Remarkably, however, loss of zonadhesin increased adhesion of mouse spermatozoa to pig, cow, and rabbit ZP but not mouse ZP. We conclude that zonadhesin mediates species-specific ZP adhesion, and Zan(-/-) males are fertile because their spermatozoa retain adhesion capability that is not species-specific. Mammalian sperm-ZP adhesion is therefore molecularly robust, and species-specific egg recognition by a protein in the sperm acrosome is conserved between invertebrates and vertebrates, even though the adhesion molecules themselves are unrelated.

Validation of two methods for the quantitative analysis of cocaine and opioids in biological matrices using LCMSMS.

The proliferation of misuse of prescription and non-prescription opioids, in recent years, has caused an opioid epidemic in the United States. Forensic toxicology laboratories often encounter implications of abuse in both driving under the influence of drugs and death investigation cases. The Virginia Department of Forensic Science has seen over a 190% increase in the number of reported opioids in death investigation cases over the past several years. Traditionally, analyses are completed by individual drug class, which subsequently requires an individual case to be evaluated using multiple analytical techniques for comprehensive analysis. To ease the impact of increasing case submissions and case complexity, two liquid chromatography-tandem mass spectrometry (LCMSMS) methods for the quantitative analysis of opioids, cocaine, and cocaine metabolites in biological matrices have undergone a fit-for-purpose validation. The methods were compared to determine the advantages and disadvantages of each analytical technique. Two sample preparation techniques, protein precipitation and solid-phase extraction, were employed for quantitative validation. Validation aspects evaluated included accuracy and precision, sensitivity, linearity, matrix effects, recovery, carryover, interferences, dilution integrity, and post-extraction stability. The accuracy of the protein precipitation method ranged from 80 ± 3% to 101 ± 10%. The accuracy of the solid-phase extraction method ranged from 88 ± 9% to 119 ± 3%. Additionally, within-run and intermediate precision was within ±20% for both extraction techniques. Although ionization suppression was noted, it was determined to have minimal effect on the methods. The newly developed methods require less sample volume and combine four analytical techniques into one method, which significantly impacts laboratory productivity.