Alteration of the Chicken Upper Respiratory Microbiota, Following H9N2 Avian Influenza Virus Infection.

Several studies have highlighted the importance of the gut microbiota in developing immunity against viral infections in chickens. We have previously shown that H9N2 avian influenza A virus (AIV) infection retards the diversity of the natural colon-associated microbiota, which may further influence chicken health following recovery from infection. The effects of influenza infection on the upper respiratory tract (URT) microbiota are largely unknown. Here, we showed that H9N2 AIV infection lowers alpha diversity indices in the acute phase of infection in the URT, largely due to the family Lactobacillaceae being highly enriched during this time in the respiratory microbiota. Interestingly, microbiota diversity did not return to levels similar to control chickens in the recovery phase after viral shedding had ceased. Beta diversity followed a similar trend following the challenge. Lactobacillus associate statistically with the disturbed microbiota of infected chickens at the acute and recovery phases of infection. Additionally, we studied age-related changes in the respiratory microbiota during maturation in chickens. From 7 to 28 days of age, species richness and evenness were observed to advance over time as the microbial composition evolved. Maintaining microbiota homeostasis might be considered as a potential therapeutic target to prevent or aid recovery from H9N2 AIV infection.

Genetic variability of oxidative stress and DNA repair genes associated with pre-treatment cancer-related fatigue in women with breast cancer.

PURPOSE: Investigate potential relationships between pre-treatment cancer-related fatigue (CRF) in women with early-stage breast cancer and variation in genes involved with oxidative stress and DNA repair. METHODS: Investigated 39 functional and tagging single nucleotide polymorphisms (SNPs) in genes involved in oxidative stress (CAT, GPX1, SEPP1, SOD1, and SOD2) and DNA repair (ERCC2, ERCC3, ERCC5, and PARP1) in a sample (N = 219) that included n = 138 postmenopausal women diagnosed with early-stage breast cancer before initiation of therapy and n = 81 age- and education-matched healthy controls. Using the Profile of Mood States Fatigue/Inertia Subscale, fatigue occurrence and severity were evaluated in both groups. Regression analysis was used to independently identify significant SNPs for three outcomes: 1) any fatigue versus no fatigue, 2) clinically meaningful versus non-clinically meaningful fatigue, and 3) fatigue severity. Using a weighted multi-SNP method, genetic risk scores (GRS) were calculated for each participant, and GRS models were constructed for each outcome. Models were adjusted for age, pain, and symptoms of depression and anxiety. RESULTS: SEPP1rs3877899, ERCC2rs238406, ERCC2rs238416, ERCC2rs3916874, and ERCC3rs2134794 were associated with fatigue occurrence and had a significant GRS model (OR = 1.317, 95%CI [1.067, 1.675], P ≤ 0.05). One SNP, SOD2rs5746136, was significant for clinically meaningful fatigue; therefore, a GRS model could not be constructed. ERCC3rs4150407, ERCC3rs4150477, and ERCC3rs2134794 were associated with fatigue severity with a significant GRS model (b = 1.010, 95%CI [1.647, 4.577], R2 = 6.9%, P ≤ 0.01). CONCLUSIONS: These results may contribute to identifying patients who are at risk of developing CRF. Oxidative stress and DNA repair biological pathways may be involved with CRF.

Brain-derived neurotrophic factor (BDNF) epigenomic modifications and brain-related phenotypes in humans: A systematic review.

Epigenomic modifications of the brain-derived neurotrophic factor (BDNF) gene have been postulated to underlie the pathogenesis of neurodevelopmental, psychiatric, and neurological conditions. This systematic review summarizes current evidence investigating the association of BDNF epigenomic modifications (DNA methylation, non-coding RNA, histone modifications) with brain-related phenotypes in humans. A novel contribution is our creation of an open access web-based application, the BDNF DNA Methylation Map, to interactively visualize specific positions of CpG sites investigated across all studies for which relevant data were available. Our literature search of four databases through September 27, 2021 returned 1701 articles, of which 153 met inclusion criteria. Our review revealed exceptional heterogeneity in methodological approaches, hindering the identification of clear patterns of robust and/or replicated results. We summarize key findings and provide recommendations for future epigenomic research. The existing literature appears to remain in its infancy and requires additional rigorous research to fulfill its potential to explain BDNF-linked risk for brain-related conditions and improve our understanding of the molecular mechanisms underlying their pathogenesis.

Epigenomic Links Between Social Determinants of Health and Symptoms: A Scoping Review.

Social determinants of health (SDoH) impact health and wellness. The link between SDoH and adverse health outcomes, including symptom occurrence and severity, may be explained by an individual's physiologic response to one or more SDoH. One potential mechanism underlying this physiologic response linking SDoH and symptoms is the dynamic epigenome. The purpose of this scoping review of the literature was to examine differential susceptibility for symptoms by identifying and summarizing research linking SDoH and symptoms through epigenomic mechanisms. PubMed was searched to identify empirical research where at least one SDoH was an independent or dependent variable, at least one symptom was investigated, and the investigation included an epigenomic measure. Of the 484 articles initially retrieved, after thorough vetting, 41 articles met eligibility. The most studied symptom was depressive symptoms followed by anxiety, cognitive function, sleep dysfunction, and pain. The most frequently studied SDoH were: 1) stress, particularly early life stress and acculturative stress; and 2) trauma, predominantly childhood trauma. DNA methylation and telomere length were the most studied epigenomic measures. Four genes (SLC6A4, BDNF, NR3C1, OXTR) had evidence from multiple studies and across methodological approaches linking SDoH to symptoms. This review supports the inclusion of epigenomic approaches to better understand the link between SDoH and symptoms and provides evidence that SDoH impact telomere length and the methylation of genes involved in neurotransmitter signaling, neuronal survival, behavior, inflammation and stress response.

Electrophysiological Measures of Swallowing Functions: A Systematic Review.

The purpose of this systematic review was to examine the application of event-related potentials (ERPs) to investigate neural processes of swallowing functions in adults with and without dysphagia. Computerized literature searches were performed from three search engines. Studies were screened using Covidence (Cochrane tool) and followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement standards (PRISMA-2009). A total of 759 studies were initially retrieved, of which 12 studies met inclusion criteria. Electrophysiological measures assessing swallowing functions were identified in two major ERP categories: (1) sensory potentials and (2) pre-motor potentials. Approximately 80% of eligible studies demonstrated strong methodological quality, although most employed a case series or case-control study design. Pharyngeal sensory-evoked potentials (PSEPs) were used to assess pharyngeal afferent cortical processing. The temporal sequence of the PSEP waveforms varied based on the sensory stimuli. PSEPs were delayed with localized scalp maps in patients with dysphagia as compared to healthy controls. The pre-motor ERPs assessed the cortical substrates involved in motor planning for swallowing, with the following major neural substrates identified: pre-motor cortex, supplementary motor area, and primary sensorimotor cortex. The pre-motor ERPs differed in amplitude for the swallow task (saliva versus liquid swallow), and the neural networks differed for cued versus non-cued task of swallowing suggesting differences in cognitive processes. This systematic review describes the application of electrophysiological measures to assess swallowing function and the promising application for furthering understanding of the neural substrates of swallowing. Standardization of protocols for use of electrophysiological measures to examine swallowing would allow for aggregation of study data to inform clinical practice for dysphagia rehabilitation.

A Preliminary Exploration of Pitch Discrimination, Temporal Sequencing, and Prosodic Awareness Skills of Children Who Participate in Different School-Based Music Curricula.

Musical training has been shown to have a positive influence on a variety of skills, including auditory-based tasks and nonmusical cognitive and executive functioning tasks; however, because previous investigations have yielded mixed results regarding the relationship between musical training and these skills, the purpose of this study was to examine and compare the auditory processing skills of children who receive focused, daily musical training with those with more limited, generalized musical training. Sixteen typically developing children (second-fourth grade) from two different schools receiving different music curricula were assessed on measures of pitch discrimination, temporal sequencing, and prosodic awareness. The results indicated significantly better scores in pitch discrimination abilities for the children receiving daily, focused musical training (School 1) compared to students attending music class only once per week, utilizing a more generalized elementary school music curriculum (School 2). The findings suggest that more in-depth and frequent musical training may be associated with better pitch discrimination abilities in children. This finding is important given that the ability to discriminate pitch has been linked to improved phonological processing skills, an important skill for developing spoken language and literacy. Future investigations are needed to determine whether the null findings for temporal sequencing and prosodic awareness can be replicated or may be different for various grades and tasks for measuring these abilities.

Interprofessional Collaborative Practice Trends Between Speech-Language Pathologists and Audiologists.

PURPOSE: This study investigated interprofessional practice (IPP) trends and attitudes between speech-language pathologists (SLP) and audiologists (AUD). METHODS: Clinical SLPs and AUDs completed an online survey, consisting of demographics, caseload trends, collaborative practice trends, and barriers to collaborative practice. RESULTS: A total of 237 participants (131 SLPs, 106 AUDs) completed an online survey. Survey results indicated that IPP is important or very important to both professions. A smaller proportion of SLPs reported collaborating with AUDs than AUDs reported collaborating with SLPs. The most frequently identified barrier to collaboration experienced by the two professions was "access." Based on their beliefs, however, the top barriers differed between the professions. Specifically, SLPs believed that "access" was the top barrier to collaboration with AUDs whereas AUDs believed that "attitudes and perceptions" was the top barrier to collaboration with SLPs. Findings from the revised Attitudes Towards Health Care Teams Scale indicated that SLPs had a significantly more positive attitude toward IPP than AUDs. CONCLUSIONS: This study indicated that although IPP is valued by SLPs and AUDs, barriers continue to limit the percentage of practitioners engaging in IPP. Although both professions experienced common barriers, their beliefs about barriers to collaboration differed.

The Contribution of Frequency Discrimination Ability to Auditory Temporal Patterning Tests in Children.

Purpose The Pitch Patterns Test (PPT) and the Duration Patterns Test (DPT) are clinical auditory processing tests that evaluate temporal patterning skills based on pitch (PPT) or duration (DPT) aspects of sound. Although temporal patterning tests are categorized under the temporal processing domain, successful performance on the PPT also relies on accurate pitch discrimination. However, the relationship between pitch discrimination ability and temporal patterning skills has not been thoroughly evaluated. This study examined the contribution of pitch discrimination ability to performance on temporal patterning in children through the use of a pitch discrimination task and the PPT. The DPT was also given as a control measure to assess temporal patterning with no pitch component. Method Thirty-two typically developing elementary school-age children (6;11-11;3 [years;months]) with normal hearing were given a series of three counterbalanced tasks: an adaptive psychophysical pitch discrimination task (difference limen for frequency [DLF]), the PPT, and the DPT. Results Correlational analysis revealed moderate correlations between DLF and PPT scores. After accounting for age, results of a linear regression analysis suggested that pitch discrimination accounts for a significant amount of variance in performance on the PPT. No significant correlation was found between DLF and DPT scores, supporting the hypothesis that the pitch task had no significant temporal patterning component contributing to the overall score. Discussion These findings indicate that pitch discrimination contributes significantly to performance on the PPT, but not the DPT, in a typically developing pediatric population. This is an important clinical consideration in both assessment and utilization of targeted therapy techniques for different clinical populations.

-----Comparison of T1-Post and FLAIR-Post MRI for identification of traumatic meningeal enhancement in traumatic brain injury patients.

Traumatic meningeal enhancement (TME) is a novel biomarker observed on post-contrast fluid-attenuated inversion recovery (FLAIR) in patients who undergo contrast-enhanced magnetic resonance imaging (MRI) after suspected traumatic brain injury (TBI). TME may be seen on acute MRI despite the absence of other trauma-related intracranial findings. In this study we compare conspicuity of TME on FLAIR post-contrast and T1 weighted imaging (T1WI) post-contrast, and investigate if TME is best detected by FLAIR post-contrast or T1WI post-contrast sequences. Subjects selected for analysis enrolled in the parent study (NCT01132937) in 2016 and underwent contrast-enhanced MRI within 48 hours of suspected TBI. Two blinded readers reviewed pairs of pre- and post-contrast T1WI and FLAIR images for presence or absence of TME. Discordant pairs between the two blinded readers were reviewed by a third reader. Cohen's kappa coefficient was used to calculate agreement. Twenty-five subjects (15 males, 10 females; median age 48 (Q1:35-Q3:62; IQR: 27)) were included. The blinded readers had high agreement for presence of TME on FLAIR (Kappa of 0.90), but had no agreement for presence of TME on T1WI (Kappa of -0.24). The FLAIR and T1WI scans were compared among all three readers and 62% of the cases positive on FLAIR could be seen on T1WI. However, 38% of the cases who were read positive on FLAIR for TME were read negative for TME on T1WI. Conspicuity of TME is higher on post-contrast FLAIR MRI than on post-contrast T1WI. TME as seen on post-contrast FLAIR MRI can aid in the identification of patients with TBI.

Inflammatory Cytokines Associate With Neuroimaging After Acute Mild Traumatic Brain Injury.

Introduction: Elevated levels of blood-based proinflammatory cytokines are linked to acute moderate to severe traumatic brain injuries (TBIs), yet less is known in acute mild (m)TBI cohorts. The current study examined whether blood-based cytokines can differentiate patients with mTBI, with and without neuroimaging findings (CT and MRI). Material and Methods: Within 24 h of a mTBI, determined by a Glasgow Coma Scale (GCS) between 13 and 15, participants (n = 250) underwent a computed tomography (CT) and magnetic resonance imaging (MRI) scan and provided a blood sample. Participants were classified into three groups according to imaging findings; (1) CT+, (2) MRI+ (CT-), (3) Controls (CT- MRI-). Plasma levels of circulating cytokines (IL-6, IL-10, TNFα), and vascular endothelial growth factor (VEGF) were measured using an ultra-sensitive immunoassay. Results: Concentrations of inflammatory cytokines (IL-6, TNFα) and VEGF were elevated in CT+, as well as MRI+ groups (p < 0.001), compared to controls, even after controlling for age, sex and cardiovascular disease (CVD)-related risk factors; hypertension, and hyperlipidemia. Post-concussive symptoms were associated with imaging groupings, but not inflammatory cytokines in this cohort. Levels of VEGF, IL-6, and TNFα differentiated patients with CT+ findings from controls, with the combined biomarker model (VEGF, IL-6, TNFα, and IL-10) showing good discriminatory power (AUC 0.92, 95% CI 0.87-0.97). IL-6 was a fair predictor of MRI+ findings compared to controls (AUC 0.70, 95% CI 0.60-0.78). Finally, the combined biomarker model discriminated patients with MRI+ from CT+ with an AUC of 0.71 (95% CI 0.62-0.80). Conclusions: When combined, IL-6, TNFα, and VEGF may provide a promising biomarker cytokine panel to differentiate mTBI patients with CT+ imaging vs. controls. Singularly, IL-6 was a fair discriminator between each of the imaging groups. Future research directions may help elucidate mechanisms related to injury severity and potentially, recovery following an mTBI.

Meningeal blood-brain barrier disruption in acute traumatic brain injury.

The meninges serve as a functional barrier surrounding the brain, critical to the immune response, and can be compromised following head trauma. Meningeal enhancement can be detected on contrast-enhanced MRI in patients presenting with acute traumatic brain injury, even when head CT is negative. Following head trauma, gadolinium-based contrast appears to extravasate from the vasculature, enhancing the dura within minutes, and later permeates the subarachnoid space. The aims of this study were to characterize the initial kinetics of the uptake of contrast agent after injury and the delayed redistribution of contrast enhancement in the subarachnoid space in hyperacute patients. Neuroimaging was obtained prospectively in two large ongoing observational studies of patients aged 18 years or older presenting to the emergency department with suspected acute head injury. Dynamic contrast-enhanced MRI studies in a cohort of consecutively enrolling patients with mild traumatic brain injury (n = 36) determined that the kinetic half-life of dural-related meningeal enhancement was 1.3 ± 0.6 min (95% enhancement within 6 min). The extravasation of contrast into the subarachnoid space was investigated in a cohort of CT negative mild traumatic brain injury patients initially imaged within 6 h of injury (hyperacute) who subsequently underwent a delayed MRI, with no additional contrast administration, several hours after the initial MRI. Of the 32 patients with delayed post-contrast imaging, 18 (56%) had conspicuous expansion of the contrast enhancement into the subarachnoid space, predominantly along the falx and superior sagittal sinus. Patients negative for traumatic meningeal enhancement on initial hyperacute MRI continued to have no evidence of meningeal enhancement on the delayed MRI. These studies demonstrate that (i) the initial enhancement of the traumatically injured meninges occurs within minutes of contrast injection, suggesting highly permeable meningeal vasculature, and that (ii) contrast in the meninges redistributes within the subarachnoid space over the period of hours, suggesting a compromise in the blood-brain and/or blood-cerebrospinal barriers. Data from the parent study indicate that up to one in two patients with mild traumatic brain injury have traumatic brain injury on acute (<48 h) MRI, with a higher prevalence seen in patients with moderate or severe traumatic brain injury. The current study's findings of traumatic meningeal enhancement and the subsequent delayed extravasation of contrast into the subarachnoid spaces indicate that a substantial percentage of patients with even mild traumatic brain injury may have a transient disruption in barriers separating the vasculature from the brain.

Structural and Functional Insights into Human Nuclear Cyclophilins.

The peptidyl prolyl isomerases (PPI) of the cyclophilin type are distributed throughout human cells, including eight found solely in the nucleus. Nuclear cyclophilins are involved in complexes that regulate chromatin modification, transcription, and pre-mRNA splicing. This review collects what is known about the eight human nuclear cyclophilins: peptidyl prolyl isomerase H (PPIH), peptidyl prolyl isomerase E (PPIE), peptidyl prolyl isomerase-like 1 (PPIL1), peptidyl prolyl isomerase-like 2 (PPIL2), peptidyl prolyl isomerase-like 3 (PPIL3), peptidyl prolyl isomerase G (PPIG), spliceosome-associated protein CWC27 homolog (CWC27), and peptidyl prolyl isomerase domain and WD repeat-containing protein 1 (PPWD1). Each "spliceophilin" is evaluated in relation to the spliceosomal complex in which it has been studied, and current work studying the biological roles of these cyclophilins in the nucleus are discussed. The eight human splicing complexes available in the Protein Data Bank (PDB) are analyzed from the viewpoint of the human spliceophilins. Future directions in structural and cellular biology, and the importance of developing spliceophilin-specific inhibitors, are considered.

The spliceosomal proteins PPIH and PRPF4 exhibit bi-partite binding.

Pre-mRNA splicing is a dynamic, multistep process that is catalyzed by the RNA (ribonucleic acid)-protein complex called the spliceosome. The spliceosome contains a core set of RNAs and proteins that are conserved in all organisms that perform splicing. In higher organisms, peptidyl-prolyl isomerase H (PPIH) directly interacts with the core protein pre-mRNA processing factor 4 (PRPF4) and both integrate into the pre-catalytic spliceosome as part of the tri-snRNP (small nuclear RNA-protein complex) subcomplex. As a first step to understand the protein interactions that dictate PPIH and PRPF4 function, we expressed and purified soluble forms of each protein and formed a complex between them. We found two sites of interaction between PPIH and the N-terminus of PRPF4, an unexpected result. The N-terminus of PRPF4 is an intrinsically disordered region and does not adopt secondary structure in the presence of PPIH. In the absence of an atomic resolution structure, we used mutational analysis to identify point mutations that uncouple these two binding sites and find that mutations in both sites are necessary to break up the complex. A discussion of how this bipartite interaction between PPIH and PRPF4 may modulate spliceosomal function is included.

A Slow Conformational Switch in the BMAL1 Transactivation Domain Modulates Circadian Rhythms.

The C-terminal transactivation domain (TAD) of BMAL1 (brain and muscle ARNT-like 1) is a regulatory hub for transcriptional coactivators and repressors that compete for binding and, consequently, contributes to period determination of the mammalian circadian clock. Here, we report the discovery of two distinct conformational states that slowly exchange within the dynamic TAD to control timing. This binary switch results from cis/trans isomerization about a highly conserved Trp-Pro imide bond in a region of the TAD that is required for normal circadian timekeeping. Both cis and trans isomers interact with transcriptional regulators, suggesting that isomerization could serve a role in assembling regulatory complexes in vivo. Toward this end, we show that locking the switch into the trans isomer leads to shortened circadian periods. Furthermore, isomerization is regulated by the cyclophilin family of peptidyl-prolyl isomerases, highlighting the potential for regulation of BMAL1 protein dynamics in period determination.

P300 Event-Related Potentials Differentiate Better Performing Individuals With Traumatic Brain Injury: A Preliminary Study of Semantic Processing.

OBJECTIVE: To measure the effect of traumatic brain injury on the cognitive processing of words, as measured by the P300, in a semantic categorization task. PARTICIPANTS: Eight adults with a history of moderate to severe traumatic brain injury and 8 age- and gender-matched controls. DESIGN: A pilot study measuring cognitive event-related potentials in response to word pairs that were either in same or different semantic categories. MAIN MEASURES: The P300 (P3b) component of the auditory event-related potential and neuropsychological assessment. RESULTS: Two patterns of P300 amplitude related to brain injury were observed. Participants with poorer performance on neuropsychological tests exhibited reduced P300 amplitude as compared to controls but showed the typical P300 parietal scalp distribution. In contrast, better performing participants demonstrated robust P300 amplitude but a substantially altered scalp distribution, characterized by the recruitment of anterior brain regions in addition to parietal activation. CONCLUSIONS: The recruitment of frontal areas after traumatic brain injury may represent compensatory neural mechanisms utilized to successfully maximize task performance. The P300 in a semantic processing paradigm may be a sensitive marker of neural plasticity that could be used to improve functional outcomes in cognitive remediation paradigms.

The Effect of Signal-to-Noise Ratio on Linguistic Processing in a Semantic Judgment Task: An Aging Study.

BACKGROUND: Aging effects on speech understanding in noise have primarily been assessed through speech recognition tasks. Recognition tasks, which focus on bottom-up, perceptual aspects of speech understanding, intentionally limit linguistic and cognitive factors by asking participants to only repeat what they have heard. On the other hand, linguistic processing tasks require bottom-up and top-down (linguistic, cognitive) processing skills and are, therefore, more reflective of speech understanding abilities used in everyday communication. The effect of signal-to-noise ratio (SNR) on linguistic processing ability is relatively unknown for either young (YAs) or older adults (OAs). PURPOSE: To determine if reduced SNRs would be more deleterious to the linguistic processing of OAs than YAs, as measured by accuracy and reaction time in a semantic judgment task in competing speech. RESEARCH DESIGN: In the semantic judgment task, participants indicated via button press whether word pairs were a semantic Match or No Match. This task was performed in quiet, as well as, +3, 0, -3, and -6 dB SNR with two-talker speech competition. STUDY SAMPLE: Seventeen YAs (20-30 yr) with normal hearing sensitivity and 17 OAs (60-68 yr) with normal hearing sensitivity or mild-to-moderate sensorineural hearing loss within age-appropriate norms. DATA COLLECTION AND ANALYSIS: Accuracy, reaction time, and false alarm rate were measured and analyzed using a mixed design analysis of variance. RESULTS: A decrease in SNR level significantly reduced accuracy and increased reaction time in both YAs and OAs. However, poor SNRs affected accuracy and reaction time of Match and No Match word pairs differently. Accuracy for Match pairs declined at a steeper rate than No Match pairs in both groups as SNR decreased. In addition, reaction time for No Match pairs increased at a greater rate than Match pairs in more difficult SNRs, particularly at -3 and -6 dB SNR. False-alarm rates indicated that participants had a response bias to No Match pairs as the SNR decreased. Age-related differences were limited to No Match pair accuracies at -6 dB SNR. CONCLUSIONS: The ability to correctly identify semantically matched word pairs was more susceptible to disruption by a poor SNR than semantically unrelated words in both YAs and OAs. The effect of SNR on this semantic judgment task implies that speech competition differentially affected the facilitation of semantically related words and the inhibition of semantically incompatible words, although processing speed, as measured by reaction time, remained faster for semantically matched pairs. Overall, the semantic judgment task in competing speech elucidated the effect of a poor listening environment on the higher order processing of words.

Recruitment Strategies and Costs Associated With Enrolling People With Insomnia and High Blood Pressure Into an Online Behavioral Sleep Intervention: A Single-Site Pilot Study.

BACKGROUND: Recruitment in clinical research is a common challenge and source of study failure. The reporting of recruitment methods and costs in hypertension trials is limited especially for smaller, single-site trials, online intervention trials, and trials using newer online recruitment strategies. OBJECTIVE: The aims of this study are to describe and examine the feasibility of newer online-e-mail recruitment strategies and traditional recruitment strategies used to enroll participants with insomnia and high blood pressure into an online behavioral sleep intervention study (Sleeping for Heart Health). METHODS: The 16 online-e-mail-based and traditional recruitment strategies used are described. Recruitment strategy feasibility was examined by study interest and enrollee yields, conversion rates, and costs (direct, remuneration, labor, and cost per enrollee). RESULTS: From August 2014 to October 2015, 183 people were screened and 58 (31.7%) enrolled in the study (51.1 ± 12.9 years, 63.8% female, 72.4% African American, 136 ± 12/88 ± 7 mm Hg, 87.9% self-reported hypertension, 67.2% self-reported antihypertensive medication use). The recruitment strategies yielding the highest enrollees were the university hospital phone waiting message system (25.4%), Craigslist (22.4%), and flyers (20.3%) at a per enrollee cost of $42.84, $98.90, and $128.27, respectively. The university hospital phone waiting message system (55.6%) and flyers (54.5%) had the highest interested participant to enrolled participant conversion rate of all recruitment strategies. CONCLUSION: Approximately 70% of all enrolled participants were recruited from the university hospital phone waiting message system, Craigslist, or flyers. Given the recruitment challenges that most researchers face, we encourage the documenting, assessing, and reporting of detailed recruitment strategies and associated recruitment costs so that other researchers may benefit.

Phosphorylation of the RNA-binding protein Dazl by MAPKAP kinase 2 regulates spermatogenesis.

Developing male germ cells are exquisitely sensitive to environmental insults such as heat and oxidative stress. An additional characteristic of these cells is their unique dependence on RNA-binding proteins for regulating posttranscriptional gene expression and translational control. Here we provide a mechanistic link unifying these two features. We show that the germ cell-specific RNA-binding protein deleted in azoospermia-like (Dazl) is phosphorylated by MAPKAP kinase 2 (MK2), a stress-induced protein kinase activated downstream of p38 MAPK. We demonstrate that phosphorylation of Dazl by MK2 on an evolutionarily conserved serine residue inhibits its interaction with poly(A)-binding protein, resulting in reduced translation of Dazl-regulated target RNAs. We further show that transgenic expression of wild-type human Dazl but not a phosphomimetic form in the Drosophila male germline can restore fertility to flies deficient in boule, the Drosophila orthologue of human Dazl. These results illuminate a novel role for MK2 in spermatogenesis, expand the repertoire of RNA-binding proteins phosphorylated by this kinase, and suggest that signaling by the p38-MK2 pathway is a negative regulator of spermatogenesis via phosphorylation of Dazl.

Age-Related Effects of Dichotic Attentional Mode on Interaural Asymmetry: An AERP Study with Independent Component Analysis.

BACKGROUND: The degree of interaural asymmetry (IA) obtained on a dichotic listening task is affected by attentional demands attributable to the mode of test administration. Previous research has shown that IA in the elderly is more influenced by increased attentional demands than young adults (YAs), but the effect of attentional mode on IA in middle-aged adults (MAs) has not been established. Auditory event-related potentials (AERPs), such as the N400, allow the evaluation of subtle differences in linguistic and cognitive processing between YAs and MAs that are imperceptible by behavioral means. PURPOSE: To determine the effect of dichotic attentional mode on IA in the N400 in YA and MA listeners. RESEARCH DESIGN: Participants listened to groups of words that consisted of a reference word followed by dichotic probe words. Participants judged whether probe words were semantically related or unrelated to the reference word. This semantic judgment task was elicited in both divided-attention (DIV) and directed-attention (DIR) modes. STUDY SAMPLE: Twenty-three YA (19-25 yr) and twenty-three MA (47-59 yr) females participated in the study. DATA COLLECTION AND ANALYSIS: Individual, as well as grand-averaged, AERP waveforms, scalp topographies, and event-related potential-image plots were analyzed. A mixed design analysis of variance was used to compare the N400 amplitude and latency response between ears in both attentional modes. Independent component analysis was used to isolate temporally overlapping neural sources that contributed to the negativity in the latency range of the N400 component. RESULTS: N400 amplitude was significantly more negative in the DIV mode than DIR in both age groups. IA differences between age groups were evident only in DIV, as indicated by an age-related shift in the direction of IA in the N400 from greater asymmetry on the right in YAs to greater asymmetry on the left in MAs. ICA revealed that the age-related difference in IA in the AERP waveform reflected differences between YAs and MAs primarily in an electroencephalographic source process consistent with attentional processing. CONCLUSIONS: IA differences between YAs and MAs were revealed in the N400 only in DIV, which was the result of an increased information-processing load. ICA successfully separated temporally overlapping EEG sources that contributed to the N400 component, allowing a refined interpretation of differences in the AERP waveform among groups.

Nuclear cyclophilins affect spliceosome assembly and function in vitro.

Cyclophilins are ubiquitously expressed proteins that bind to prolines and can catalyse cis/trans isomerization of proline residues. There are 17 annotated members of the cyclophilin family in humans, ubiquitously expressed and localized variously to the cytoplasm, nucleus or mitochondria. Surprisingly, all eight of the nuclear localized cyclophilins are found associated with spliceosomal complexes. However, their particular functions within this context are unknown. We have therefore adapted three established assays for in vitro pre-mRNA splicing to probe the functional roles of nuclear cyclophilins in the context of the human spliceosome. We find that four of the eight spliceosom-associated cyclophilins exert strong effects on splicing in vitro. These effects are dose-dependent and, remarkably, uniquely characteristic of each cyclophilin. Using both qualitative and quantitative means, we show that at least half of the nuclear cyclophilins can act as regulatory factors of spliceosome function in vitro. The present work provides the first quantifiable evidence that nuclear cyclophilins are splicing factors and provides a novel approach for future work into small molecule-based modulation of pre-mRNA splicing.