Polyamines Control eIF5A Hypusination, TFEB Translation, and Autophagy to Reverse B Cell Senescence.

Failure to make adaptive immune responses is a hallmark of aging. Reduced B cell function leads to poor vaccination efficacy and a high prevalence of infections in the elderly. Here we show that reduced autophagy is a central molecular mechanism underlying immune senescence. Autophagy levels are specifically reduced in mature lymphocytes, leading to compromised memory B cell responses in old individuals. Spermidine, an endogenous polyamine metabolite, induces autophagy in vivo and rejuvenates memory B cell responses. Mechanistically, spermidine post-translationally modifies the translation factor eIF5A, which is essential for the synthesis of the autophagy transcription factor TFEB. Spermidine is depleted in the elderly, leading to reduced TFEB expression and autophagy. Spermidine supplementation restored this pathway and improved the responses of old human B cells. Taken together, our results reveal an unexpected autophagy regulatory mechanism mediated by eIF5A at the translational level, which can be harnessed to reverse immune senescence in humans.

Stimulating and toxic effect of chromium on growth and photosynthesis of a marine chlorophyte.

Marine phytoplankton can interchange trace metals in various biochemical functions, particularly under metal-limiting conditions. Here, we investigate the stimulating and toxicity effect of chromium (Cr) on a marine Chlorophyceae Osetreococcus tauri under Fe-replete and Fe-deficient conditions. We determined the growth, photosynthesis, and proteome expressions of Osetreococcus tauri cultured under different Cr and Fe concentrations. In Fe-replete conditions, the presence of Cr(VI) stimulated significantly the growth rate and the maximum yield of photochemistry of photosystem II (Fv /Fm ) of the phytoplankton, while the functional absorption cross-section of photosystem II (σPSII ) did not change. Minor additions of Cr(VI) partially rescued phytoplankton growth under Fe-limited conditions. Proteomic analysis of this alga grown in Fe-replete normal and Fe-replete with Cr addition media (10 µM Cr) showed that the presence of Cr significantly decreased the expression of phosphate-transporting proteins and photosynthetic proteins, while increasing the expression of proteins related to carbon assimilation. Cr can stimulate the growth and photosynthesis of O. tauri, but the effects are dependent on both the Cr(VI) concentration and the availability of Fe. The proteomic results further suggest that Cr(VI) addition might significantly increase starch production and carbon fixation.

Smartphone addiction prevalence, patterns of use, and experienced musculoskeletal discomfort during the COVID-19 pandemic in a general Iranian population.

BACKGROUND: Smartphone usage is an essential everyday tool in Iran, however problematic use has escalated and become a concern for the Iranian health policy system, particularly during and following the COVID-19 Pandemic. This study's aim was investigation of the prevalence of smartphone addiction, patterns of use, and the relationship to specific demographic characteristics and associated musculoskeletal disorders during the COVID-19 pandemic. METHODS: A descriptive-analytical correlational study recruited participants from a population of convenience (n = 2344) who were smartphone owners with > 1 year of use. For demographic information an electronic self-report questionnaire collected age, sex, marital status, usage for daily hours, and patterns. To assess addiction levels, the 'Smartphone Addiction Scale-short version' (SAS-SV) patient-reported outcome measure was used (cut-off = 31). For experienced discomfort, the Extended Nordic Musculoskeletal Questionnaire (ENMQ) was used. RESULTS: The participants (female = 66.6%, n = 1561, mean age = 29.07 ± 12.34 years, range 6-60 years) smartphone use averaged 5.75 ± 3.44 h/day. The general prevalence of smartphone addiction was 46.16% (females = 46.06%, males = 46.36%; married = 44.5%, single = 47.63%). School students had the greatest addiction (53.2%) and those with a higher education to or above a Master's degree were the lowest (39.38%). The highest pattern of use was for social networks at 89.1% of participants (female = 88.34%, male = 90.54%). The areas of highest reported discomfort were the eyes (43.5%) and neck (43.3%). A significant correlation was found between smartphone addiction and hours of daily usage, and the amount of usage increased during the COVID-19 pandemic period. CONCLUSION: A high level of smartphone addiction in the Iranian population was found to have occurred during the COVID-19 pandemic. Those most affected were unmarried individuals and school students, with the predominant areas being the eyes and neck. Health decision-makers should consider these findings when developing recommendations and plans for public health, particularly those focused on students.

Translation, cultural adaptation and assessment of psychometrics properties of the Extended Version of the Nordic Musculoskeletal Questionnaire (NMQ-E) in Persian language speaking people.

BACKGROUND: To translate and cross-culturally adapt the Extended Version of the Nordic Musculoskeletal Questionnaire (NMQ-E) into Persian (NMQ-E-P) and evaluate the psychometric properties in a general population with different occupational tasks across nine body regions. METHODS: This cross-sectional study was designed according to the standard guidelines and the COSMIN checklist. The NMQ-E-P was achieved through forward and backward translation methods and consensus to produce the final draft. A Persian-speaking population (n = 571, age 38.24 ± 7.65 years, female = 46.2%) was recruited from industries and office workers with three occupational task inclusion criteria: assembly, office, and lifting. Psychometric properties included validity for face (from confirmed clarity, simplicity, and readability), content (via the content validity index); and construct (through known group validity); additionally, the properties of internal consistency (Cronbach's α); and test-retest reliability (Kappa coefficient of agreement) were considered. RESULTS: No significant issues during the translation process were found. The NMQ-E-P showed adequate internal consistency for all regions (α ≥ 0.87). The test-retest reliability was examined with Kappa agreement correlation coefficient and all items, except ankle regions, showed very good agreements (Kappa coefficient = 0.87-1.0). Excellent ICC values were obtained for quantitative variables (ICC > 0.88) and good construct validity was revealed (p < 0.001). CONCLUSION: The Persian version of the NMQ-E has very good validity and reliability and can be used by researchers and professionals to evaluate the prevalence of MSDs in nine body regions simultaneously.

Developing a shortened spine functional index (SFI-10) for patients with sub-acute/chronic spinal disorders: a cross-sectional study.

BACKGROUND: Brief whole-spine patient-reported outcome measures (PROMs) provide regional solutions and future directions for quantifying functional status, evidence, and effective interventions. The whole-spine regional Spine Functional Index (SFI-25) is used internationally in clinical and scientific contexts to assess general sub-acute/chronic spine populations. However, to improve structural validity and practicality a shortened version is recommended. This study developed a shortened-SFI from the determined optimal number of item questions that: correlated with criteria PROMs being highly with whole-spine, moderately with regional-spine, condition-specific and patient-specific, and moderately-low with general-health and pain; retained one-dimensional structural validity and high internal consistency; and improved practicality to reduce administrative burden. METHODS: A cross-sectional study (n = 505, age = 18-87 yrs., average = 40.3 ± 10.1 yrs) of sub-acute/chronic spine physiotherapy outpatients from an international sample of convenience. Three shortened versions of the original SFI-25 were developed using 1) qualitative 'content-retention' methodology, 2) quantitative 'factorial' methodology, and 3) quantitative 'Rasch' methodology, with a fourth 'random' version produced as a comparative control. The clinimetric properties were established for structural validity with exploratory (EFA) and confirmatory (CFA) factorial analysis, and Rasch analysis. Criterion validity used the: whole-spine SFI-25 and Functional Rating Index (FRI); regional-spine Neck Disability Index (NDI), Oswestry Disability Index (ODI), and Roland Morris Questionnaire (RMQ), condition-specific Whiplash Disability Questionnaire (WDQ); and patient-specific functional scale (PSFS); and determined floor/ceiling effect. A post-hoc pooled international sub-acute/chronic spine sample (n = 1433, age = 18-91 yrs., average = 42.0 ± 15.7 yrs) clarified the findings and employed the general-health EuroQuol-Index (EQ-5D), and 11-point Pain Numerical Rating Scale (P-NRS) criteria. RESULTS: A 10-item SFI retained structural validity with optimal practicality requiring no computational aid. The SFI-10 concept-retention-version demonstrated preferred criterion validity with whole-spine criteria (SFI-25 = 0.967, FRI = 0.810) and exceeded cut-off minimums with regional-spine, condition-specific, and patient-specific measures. An unequivocal one-dimensional structure was determined. Internal consistency was satisfactory (α = 0.80) with no floor/ceiling effect. Post-hoc analysis of the international sample confirmed these findings. CONCLUSION: The SFI-10 qualitative concept-retention version was preferred to quantitative factorial and Rasch versions, demonstrated structural and criterion validity, and preferred correlation with criteria measures. Further longitudinal research is required for reliability, error, and responsiveness, plus an examination of the practical characteristics of readability and administrative burden.

Drosophila Sex Peptide controls the assembly of lipid microcarriers in seminal fluid.

Seminal fluid plays an essential role in promoting male reproductive success and modulating female physiology and behavior. In the fruit fly, Drosophila melanogaster, Sex Peptide (SP) is the best-characterized protein mediator of these effects. It is secreted from the paired male accessory glands (AGs), which, like the mammalian prostate and seminal vesicles, generate most of the seminal fluid contents. After mating, SP binds to spermatozoa and is retained in the female sperm storage organs. It is gradually released by proteolytic cleavage and induces several long-term postmating responses, including increased ovulation, elevated feeding, and reduced receptivity to remating, primarily signaling through the SP receptor (SPR). Here, we demonstrate a previously unsuspected SPR-independent function for SP. We show that, in the AG lumen, SP and secreted proteins with membrane-binding anchors are carried on abundant, large neutral lipid-containing microcarriers, also found in other SP-expressing Drosophila species. These microcarriers are transferred to females during mating where they rapidly disassemble. Remarkably, SP is a key microcarrier assembly and disassembly factor. Its absence leads to major changes in the seminal proteome transferred to females upon mating. Males expressing nonfunctional SP mutant proteins that affect SP's binding to and release from sperm in females also do not produce normal microcarriers, suggesting that this male-specific defect contributes to the resulting widespread abnormalities in ejaculate function. Our data therefore reveal a role for SP in formation of seminal macromolecular assemblies, which may explain the presence of SP in Drosophila species that lack the signaling functions seen in Dmelanogaster.

Translation, cross-cultural adaptation, and validation of the 10-item spine functional index (SFI-10) in the Brazilians with musculoskeletal spine disorders.

PURPOSE: To translate and cross-culturally adapt the Spine Functional Index (SFI) into Brazilian Portuguese (SFI-Br) in individuals with musculoskeletal spine disorders. METHODS: Participants (n=194) answered the Numerical Pain Rating Scale (NPRS), 36-item Short-Form Health Survey (SF-36), Roland-Morris Disability Questionnaire for General Pain (RMDQ-g), and SFI-25 incorporating the SFI-10. Structural validity, from confirmatory factor analysis (CFA), used comparative fit index (CFI), Tucker-Lewis index (TLI), root mean square error of approximation (RMSEA), and chi-square/degrees of freedom (DF). The best structure was considered from the lower values of the Akaike Information Criterion (AIC) and Bayesian Information Criterion (BIC). Construct and criterion validity used Spearman's correlation coefficient (rho). Internal consistency used Cronbach's alpha, reliability used intraclass correlation coefficient (ICC2,1), with ceiling and floor effects determined. Error used the standard error of the measurement (SEM) and minimal detectable change, 90% level (MDC90). RESULTS: Adequate fit indices demonstrated an unequivocal one-factor structure only for the SFI-10 (chi-square/DF <3.00, CFI and TLI >0.90, RMSEA <0.08). The SFI-10-Br correlation was high with the SFI-Br (rho=0.914, p<0.001), moderate for the RMDQ-g (rho=-0.78), SF-36 functional capacity domain (rho=0.718) and NPRS (rho=-0.526); and adequate for the remaining SF-36 domains (rho>0.30). Test-retest reliability (ICC2,1=0.826) and internal consistency (alpha=0.864) were high. No ceiling or floor effects were observed, and error was satisfactory (SEM=9.08%, MDC90=25.15%). CONCLUSION: The SFI Brazilian version was successfully produced with the 10-item version showing an unequivocal one-factor structure, high construct and criterion validity, reliability, internal consistency, and satisfactory error. Further research on responsiveness is required.

Perfusate Proteomes Provide Biological Insight Into Oxygenated Versus Standard Hypothermic Machine Perfusion in Kidney Transplantation.

OBJECTIVE: To provide mechanistic insight into key biological alterations in donation after circulatory death kidneys during continuous pefusion we performed mass spectrometry profiling of perfusate samples collected during a phase 3 randomized double-blind paired clinical trial of hypothermic machine perfusion with and without oxygen (COMPARE). BACKGROUND: Despite the clinical benefits of novel perfusion technologies aiming to better preserve donor organs, biological processes that may be altered during perfusion have remained largely unexplored. The collection of serial perfusate samples during the COMPARE clinical trial provided a unique resource to study perfusate proteomic profiles, with the hypothesis that in-depth profiling may reveal biologically meaningful information on how donor kidneys benefit from this intervention. METHODS: Multiplexed liquid chromatography-tandem mass spectrometry was used to obtain a proteome profile of 210 perfusate samples. Partial least squares discriminant analysis and multivariate analysis involving clinical and perfusion parameters were used to identify associations between profiles and clinical outcomes. RESULTS: Identification and quantitation of 1716 proteins indicated that proteins released during perfusion originate from the kidney tissue and blood, with blood-based proteins being the majority. Data show that the overall hypothermic machine perfusion duration is associated with increasing levels of a subgroup of proteins. Notably, high-density lipoprotein and complement cascade proteins are associated with 12-month outcomes, and blood-derived proteins are enriched in the perfusate of kidneys that developed acute rejection. CONCLUSIONS: Perfusate profiling by mass spectrometry was informative and revealed proteomic changes that are biologically meaningful and, in part, explain the clinical observations of the COMPARE trial.

Relationships between cervical sagittal posture, muscle endurance, joint position sense, range of motion and level of smartphone addiction.

BACKGROUND: Frequent smartphone use in a pathological way forces the user to adopt a compromised posture. This gradually results in changes to both the postural and musculoskeletal systems. This study's objectives were evaluation of head posture, muscle endurance, neck range of motion (ROM) and joint position sense in two separate smartphone user groups, one 'Addicted', the other 'Non-Addicted'. METHODS: A sample of convenience (n = 60) was recruited from medical students (age 24.57 ± 4.38, 53.3% male) with a history of smartphones use > 2 h/day for 1-year. Based on the cut-off values of the smartphone addiction scale-short version (SAS-SV), participants were entered into each group (cut-off for male ≥ 31, female ≥ 33). Neck muscle endurance time, joint position error and cervical ROM, along with forward head posture parameters of craniovertebral angle (CVA), shoulder angle (SA), sagittal head angle (SHA) and forward head distance (FHD)) were evaluated. A Mann-Whitney test and Spearman correlation coefficient were used to determine the difference between groups and the correlations between variables. RESULTS: The difference between 'Addicted' and 'Non-Addicted' groups was confirmed by the values for SAS-SV scores (25.23 ± 5.5 versus 43.9 ± 6.61) (p < 0.001). There were statistically significant differences between groups for the CVA and FHD parameters (p < 0.001). Further, the neck extensor muscle endurance (97 ± 3.79 versus 74.86 ± 2.23 s), was significantly different between groups (p = 0.010) but not after Bonferroni correction. There was no notable difference in the neck flexor muscle endurance, joint position error, SA, and SHA parameters between groups (p > 0.05). CONCLUSIONS: There is a positive correlation between smartphone addiction and both decreased extensor muscle endurance and changes in neck postural alignment.

Male reproductive aging arises via multifaceted mating-dependent sperm and seminal proteome declines, but is postponable in Drosophila.

Declining ejaculate performance with male age is taxonomically widespread and has broad fitness consequences. Ejaculate success requires fully functional germline (sperm) and soma (seminal fluid) components. However, some aging theories predict that resources should be preferentially diverted to the germline at the expense of the soma, suggesting differential impacts of aging on sperm and seminal fluid and trade-offs between them or, more broadly, between reproduction and lifespan. While harmful effects of male age on sperm are well known, we do not know how much seminal fluid deteriorates in comparison. Moreover, given the predicted trade-offs, it remains unclear whether systemic lifespan-extending interventions could ameliorate the declining performance of the ejaculate as a whole. Here, we address these problems using Drosophila melanogaster. We demonstrate that seminal fluid deterioration contributes to male reproductive decline via mating-dependent mechanisms that include posttranslational modifications to seminal proteins and altered seminal proteome composition and transfer. Additionally, we find that sperm production declines chronologically with age, invariant to mating activity such that older multiply mated males become infertile principally via reduced sperm transfer and viability. Our data, therefore, support the idea that both germline and soma components of the ejaculate contribute to male reproductive aging but reveal a mismatch in their aging patterns. Our data do not generally support the idea that the germline is prioritized over soma, at least, within the ejaculate. Moreover, we find that lifespan-extending systemic down-regulation of insulin signaling results in improved late-life ejaculate performance, indicating simultaneous amelioration of both somatic and reproductive aging.

Integrated Plasma and Tissue Proteomics Reveals Attractin Release by Intraluminal Thrombus of Abdominal Aortic Aneurysms and Improves Aneurysm Growth Prediction in Humans.

OBJECTIVE: Discovery of novel biomarkers for AAA growth prediction. BACKGROUND: Novel biomarker of AAA growth is a recognized priority in research. Our prior work implicated intraluminal thrombus (ILT) in AAAs to be a potential source of systemic mediators during AAA progression. Here we applied a mass spectrometry proteomics pipeline to discover novel biomarkers for AAA growth prediction. METHODS: Patients were prospectively recruited. Plasma samples were collected at baseline (n = 62). AAA growth was recorded at 12 months. In Experiment 1, plasma samples from the fastest and slowest growth patients (n = 10 each) were compared. In Experiment 2, plasma samples were collected before and at 10-12 weeks after surgery (n = 29). In Experiment 3, paired ILT and omental biopsies were collected intra-operatively during open surgical repair (n = 3). In Experiment 4, tissue secretome was obtained from ex-vivo culture of these paired tissue samples. Samples were subjected to a liquid chromatography tandem mass spectrometry workflow to discover novel biomarkers. RESULTS: We discovered 3 proteins that are: (i) present in ILT; (ii) released by ILT; (iii) reduced in circulation after AAA surgery; (iv) differs between fast and slow growth AAAs. One of these is Attractin. Plasma Attractin correlates significantly with future AAA growth (Spearman r = 0.35, P < 0.005). Using Attractin and AAA diameter as input variables, the area under receiver operating characteristics for predicting no growth and fast growth or AAA at 12 months is 85% and 76%, respectively. CONCLUSION: We show that ILT of AAAs releases mediators during the natural history of AAA growth. These are novel biomarkers for AAA growth prediction in humans.

Smartphone addiction in children: patterns of use and musculoskeletal discomfort during the COVID-19 pandemic in Iran.

BACKGROUND: Smartphone use has increased significantly, especially during the period of global pandemic caused by the novel SARS-CoV2 coronavirus (COVID-19). Concurrently, smartphone addiction is a growing social problem in children and adolescents with the consequence of adverse health outcomes. This study assessed the prevalence of smartphone addiction, patterns of use, and the experienced body-region discomfort among Iranian school students during the COVID-19 pandemic. METHODS: A cross-sectional study with students from grades 1-9 recruited n = 585 participants (mean age = 14.49 (2.26 years); female = 65.8%). Data were collected from parents and students through the online 'Smartphone addiction scale-short version' (SAS-SV), self-reported demographic questionnaires, and extracts of the Nordic musculoskeletal questionnaire for the evaluation of musculoskeletal disorders. RESULTS: The prevalence rate of smartphone addiction (53.3%) was relatively high in the overall sample. Participants spent 6.85 (4.62) hours per day on their smartphones, which had increased 53.86% relative to the pre-pandemic period. The primary smartphone uses were for social networking (77.9%), web-surfing (53.3%), and camera activities (50.9%). There was a positive correlation between smartphone addiction as assessed with the SAS-SV and daily use time (r = 0.34, p < 0.001), and the percentage of change relative to the pre-pandemic period (r = 0.26, p < 0.001). Discomfort related to smartphone use was mostly reported as present in the eyes (39.7%) and neck (39.1%). A positive correlation was found (p < 0.001) between smartphone addiction and discomfort in the eyes, neck, wrists, shoulders, and upper-back. CONCLUSION: The more frequent usage of smartphones by students during the Covid-19 pandemic were associated predominantly with discomfort to the eyes and neck. Parents should consider the complications of musculoskeletal and postural changes during the child's future years and pay particular attention to the individual's patterns of smartphone use with an emphasis on posture and usage that reduces discomfort to the eyes and the musculoskeletal system, particularly the neck.

BMP signaling inhibition in Drosophila secondary cells remodels the seminal proteome and self and rival ejaculate functions.

Seminal fluid proteins (SFPs) exert potent effects on male and female fitness. Rapidly evolving and molecularly diverse, they derive from multiple male secretory cells and tissues. In Drosophila melanogaster, most SFPs are produced in the accessory glands, which are composed of ∼1,000 fertility-enhancing "main cells" and ∼40 more functionally cryptic "secondary cells." Inhibition of bone morphogenetic protein (BMP) signaling in secondary cells suppresses secretion, leading to a unique uncoupling of normal female postmating responses to the ejaculate: refractoriness stimulation is impaired, but offspring production is not. Secondary-cell secretions might therefore make highly specific contributions to the seminal proteome and ejaculate function; alternatively, they might regulate more global-but hitherto undiscovered-SFP functions and proteome composition. Here, we present data that support the latter model. We show that in addition to previously reported phenotypes, secondary-cell-specific BMP signaling inhibition compromises sperm storage and increases female sperm use efficiency. It also impacts second male sperm, tending to slow entry into storage and delay ejection. First male paternity is enhanced, which suggests a constraint on ejaculate evolution whereby high female refractoriness and sperm competitiveness are mutually exclusive. Using quantitative proteomics, we reveal changes to the seminal proteome that surprisingly encompass alterations to main-cell-derived proteins, indicating important cross-talk between classes of SFP-secreting cells. Our results demonstrate that ejaculate composition and function emerge from the integrated action of multiple secretory cell types, suggesting that modification to the cellular make-up of seminal-fluid-producing tissues is an important factor in ejaculate evolution.

Divergent allocation of sperm and the seminal proteome along a competition gradient in Drosophila melanogaster.

Sperm competition favors large, costly ejaculates, and theory predicts the evolution of allocation strategies that enable males to plastically tailor ejaculate expenditure to sperm competition threat. While greater sperm transfer in response to a perceived increase in the risk of sperm competition is well-supported, we have a poor understanding of whether males (i) respond to changes in perceived intensity of sperm competition, (ii) use the same allocation rules for sperm and seminal fluid, and (iii) experience changes in current and future reproductive performance as a result of ejaculate compositional changes. Combining quantitative proteomics with fluorescent sperm labeling, we show that Drosophila melanogaster males exercise independent control over the transfer of sperm and seminal fluid proteins (SFPs) under different levels of male-male competition. While sperm transfer peaks at low competition, consistent with some theoretical predictions based on sperm competition intensity, the abundance of transferred SFPs generally increases at high competition levels. However, we find that clusters of SFPs vary in the directionality and sensitivity of their response to competition, promoting compositional change in seminal fluid. By tracking the degree of decline in male mating probability and offspring production across successive matings, we provide evidence that ejaculate compositional change represents an adaptive response to current sperm competition, but one that comes at a cost to future mating performance. Our work reveals a previously unknown divergence in ejaculate component allocation rules, exposes downstream costs of elevated ejaculate investment, and ultimately suggests a central role for ejaculate compositional plasticity in sexual selection.

Quantitative Proteomics Identification of Seminal Fluid Proteins in Male Drosophila melanogaster.

Seminal fluid contains some of the fastest evolving proteins currently known. These seminal fluid proteins (Sfps) play crucial roles in reproduction, such as supporting sperm function, and particularly in insects, modifying female physiology and behavior. Identification of Sfps in small animals is challenging, and often relies on samples taken from the female reproductive tract after mating. A key pitfall of this method is that it might miss Sfps that are of low abundance because of dilution in the female-derived sample or rapid processing in females. Here we present a new and complementary method, which provides added sensitivity to Sfp identification. We applied label-free quantitative proteomics to Drosophila melanogaster, male reproductive tissue - where Sfps are unprocessed, and highly abundant - and quantified Sfps before and immediately after mating, to infer those transferred during copulation. We also analyzed female reproductive tracts immediately before and after copulation to confirm the presence and abundance of known and candidate Sfps, where possible. Results were cross-referenced with transcriptomic and sequence databases to improve confidence in Sfp detection. Our data were consistent with 125 previously reported Sfps. We found nine high-confidence novel candidate Sfps, which were both depleted in mated versus, unmated males and identified within the reproductive tract of mated but not virgin females. We also identified 42 more candidates that are likely Sfps based on their abundance, known expression and predicted characteristics, and revealed that four proteins previously identified as Sfps are at best minor contributors to the ejaculate. The estimated copy numbers for our candidate Sfps were lower than for previously identified Sfps, supporting the idea that our technique provides a deeper analysis of the Sfp proteome than previous studies. Our results demonstrate a novel, high-sensitivity approach to the analysis of seminal fluid proteomes, whose application will further our understanding of reproductive biology.

A comparative evaluation of telehealth and direct assessment when screening for spasticity in residents of two long-term care facilities.

OBJECTIVE: To evaluate the performance of telehealth as a screening tool for spasticity compared to direct patient assessment in the long-term care setting. DESIGN: Cross-sectional, observational study. SETTING: Two long-term care facilities: a 140-bed veterans' home and a 44-bed state home for individuals with intellectual and developmental disabilities. SUBJECTS: Sixty-one adult residents of two long-term care facilities (aged 70.1 ± 16.2 years) were included in this analysis. Spasticity was identified in 43% of subjects (Modified Ashworth Scale rating mode = 2). Contributing diagnoses included traumatic brain injury, spinal cord injury, birth trauma, stroke, cerebral palsy, and multiple sclerosis. MAIN MEASURES: Movement disorders neurologists conducted in-person examinations to determine whether spasticity was present (reference standard) and also evaluated subjects with spasticity using the Modified Ashworth Scale. Telehealth screening examinations, facilitated by a bedside nurse, were conducted remotely by two teleneurologists using a three-question screening tool. Telehealth screening determinations of spasticity were compared to the reference standard determination to calculate sensitivity, specificity, and the area under the curve (AUC) in receiver operating characteristics. Teleneurologist agreement was evaluated using Cohen's kappa. RESULTS: Teleneurologist 1 had a specificity of 89% and sensitivity of 65% to identify the likely presence of spasticity (n = 61; AUC = 0.770). Teleneurologist 2 showed 100% specificity and 82% sensitivity (n = 16; AUC = 0.909). There was almost perfect agreement between the two examiners at 94% (kappa = 0.875, 95% CI: 0.640-1.000). CONCLUSION: Telehealth may provide a useful, efficient method of identifying residents of long-term care facilities that likely need referral for spasticity evaluation.

An ALS-linked mutation in TDP-43 disrupts normal protein interactions in the motor neuron response to oxidative stress.

TDP-43 pathology is a key feature of amyotrophic lateral sclerosis (ALS), but the mechanisms linking TDP-43 to altered cellular function and neurodegeneration remain unclear. We have recently described a mouse model in which human wild-type or mutant TDP-43 are expressed at low levels and where altered stress granule formation is a robust phenotype of TDP-43M337V/- expressing cells. In the present study we use this model to investigate the functional connectivity of human TDP-43 in primary motor neurons under resting conditions and in response to oxidative stress. The interactome of human TDP-43WT or TDP-43M337V was compared by mass spectrometry, and gene ontology enrichment analysis identified pathways dysregulated by the M337V mutation. We found that under normal conditions the interactome of human TDP-43WT was enriched for proteins involved in transcription, translation and poly(A)-RNA binding. In response to oxidative stress, TDP-43WT recruits proteins of the endoplasmic reticulum and endosomal-extracellular transport pathways, interactions which are reduced in the presence of the M337V mutation. Specifically, TDP-43M337V impaired protein-protein interactions involved in stress granule formation including reduced binding to the translation initiation factors Poly(A)-binding protein and Eif4a1 and the endoplasmic reticulum chaperone Grp78. The M337V mutation also affected interactions involved in endosomal-extracellular transport and this this was associated with reduced extracellular vesicle secretion in primary motor neurons from TDP-43M337V/- mice and in human iPSCs-derived motor neurons. Taken together, our analysis highlights a TDP-43 interaction network in motor neurons and demonstrates that an ALS associated mutation may alter the interactome to drive aberrant pathways involved in the pathogenesis of ALS.

CSF extracellular vesicle proteomics demonstrates altered protein homeostasis in amyotrophic lateral sclerosis.

BACKGROUND: Extracellular vesicles (EVs) released by neurons and glia reach the cerebrospinal fluid (CSF). Studying the proteome of CSF-derived EVs offers a novel perspective on the key intracellular processes associated with the pathogenesis of the neurodegenerative disease amyotrophic lateral sclerosis (ALS) and a potential source from which to develop biomarkers. METHODS: CSF EVs were extracted using ultrafiltration liquid chromatography from ALS patients and controls. EV size distribution and concentration was measured using nanoparticle tracking analysis and liquid chromatography-tandem mass spectrometry proteomic analysis performed. RESULTS: CSF EV concentration and size distribution did not differ between ALS and control groups, nor between a sub-group of ALS patients with or without an associated hexanucleotide repeat expansion (HRE) in C9orf72. Univariate proteomic analysis identified downregulation of the pentameric proteasome-like protein Bleomycin hydrolase in ALS patients, whilst Gene Ontology enrichment analysis demonstrated downregulation of proteasome core complex proteins (8/8 proteins, normalized enrichment ratio -1.77, FDR-adjusted p = 0.057) in the ALS group. The sub-group of ALS patients associated with the C9orf72 HRE showed upregulation in Ubiquitin-like modifying-activating protein 1 (UBA1) compared to non-C9orf72 cases. CONCLUSIONS: Proteomic analysis of CSF EVs in ALS detects intracellular alterations in protein homeostatic mechanisms, previously only identified in pathological tissues. This supports the wider use of CSF EVs as a source of novel biomarkers reflecting key and potentially druggable pathological intracellular pathway alterations in ALS.

Two-dimensional 67Zn HYSCORE spectroscopy reveals that a Zn-bacteriochlorophyll aP' dimer is the primary donor (P840) in the type-1 reaction centers of Chloracidobacterium thermophilum.

Chloracidobacterium (C.) thermophilum is a microaerophilic, chlorophototrophic species in the phylum Acidobacteria that uses homodimeric type-1 reaction centers (RC) to convert light energy into chemical energy using (bacterio)chlorophyll ((B)Chl) cofactors. Pigment analyses show that these RCs contain BChl aP, Chl aPD, and Zn2+-BChl aP' in the approximate ratio 7.1 : 5.4 : 1. However, the functional roles of these three different Chl species are not yet fully understood. It was recently demonstrated that Chl aPD is the primary electron acceptor. Because Zn2+-(B)Chl aP' is present at low abundance, it was suggested that the primary electron donor might be a dimer of Zn2+-BChl aP' molecules. In this study, we utilize isotopic enrichment and high-resolution two-dimensional (2D) 14N and 67Zn hyperfine sublevel correlation (HYSCORE) spectroscopy to demonstrate that the primary donor cation, P840+, in the C. thermophilum RC is indeed a Zn2+-BChl aP' dimer. Density functional theory (DFT) calculations and the measured electron-nuclear hyperfine parameters of P840+ indicate that the electron spin density on P840+ is distributed nearly symmetrically over two Zn2+-(B)Chl aP' molecules as expected in a homodimeric RC. To our knowledge this is the only example of a photochemical RC in which the Chl molecules of the primary donor are metallated differently than those of the antenna.

Moral Distress of Clinicians in Canadian Pediatric and Neonatal ICUs.

OBJECTIVE: To quantify moral distress in neonatal ICU and PICU clinicians and to identify associated factors. DESIGN: A national cross-sectional survey of clinicians working in an neonatal ICU or PICU. Moral distress was assessed with the Moral Distress Scale-Revised and by self-rating. Depersonalization was assessed on the subscale of the Maslach Burnout Inventory. Respondents reported their attendance at each of six hospital supports that may serve to mitigate moral distress in frontline staff. Analyses compared outcomes across respondent characteristics and hierarchical linear regression evaluated individual, ICU, hospital, and regional effects. SETTING: Eligible ICUs were PICUs and level-3 neonatal ICUs in Canada. SUBJECTS: Eligible participants had worked in the participating ICU for more than 3 months. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: We identified 54 eligible ICUs from 31 hospitals. Forty-nine Canadian neonatal ICUs and PICUs (91%) contributed 2,852 complete responses for a 45.2% response rate. Most respondents were nurses (64.9%) or from a neonatal ICU (66.5%). The median and interquartile range Moral Distress Scale-Revised were 79 (52-113); 997 respondents (34.2%) had Moral Distress Scale-Revised scores greater than or equal to 100, and 234 respondents (8.3%) strongly agreed that work caused them significant moral distress. Nurses had a median (interquartile range) Moral Distress Scale-Revised score of 85 (57-121), 19 points higher than physicians and 8 points higher than respiratory therapists (p < 0.0001). Moral Distress Scale-Revised scores increased from 53 (35-79) for those working in ICU less than 1 year to 83 (54-120) in those working in ICU more than 30 years (p < 0.0001); 22.5% reported high degrees of depersonalization, which was associated with moral distress (p < 0.0001). Variability in Moral Distress Scale-Revised scores was explained by individual-level (92%), hospital-level (5%), and ICU-level effects (1%). Frequency of participation in potentially mitigating hospital supports had small effects (< 10 points) on mean Moral Distress Scale-Revised scores. CONCLUSIONS: Moral distress is common in clinicians working in ICUs for children. Addressing moral distress will require interventions tailored to individuals in higher-risk groups.