Tendon cell ciliary length as a biomarker of in situ cytoskeletal tensional homeostasis

To determine if tendon cell ciliary length could be used as a biomarker of cytoskeletal tensional homeostasis, 20 mm long adult rat tail tendons were placed in double artery clamps set 18 mm apart to create a 10% laxity. The tendons were allowed to contract over a 7 day period under culture conditions. At 0, 1, 5, and 7 days the tendon cell cilia were stained and ciliary length measured using confocal imaging. There was a significant (p<0.001) increase in ciliary length at 1 day. At day 5 (when the tendon became visibly taut) there was a significant (p<0.001) decrease in ciliary length compared to day 1. By day 7 the tendon remained taut and ciliary length returned to day zero values (p=0.883). These results suggest that cilia length reflects the local mechanobiological environment of tendon cells and could be used as a potential in situ biomarker of altered cytoskeletal tensional homeostasi

How people get into mental health services: Stories of choice, coercion and "muddling through" from "first-timers"

Previous work examining how individuals enter mental health treatment comes either from the health services utilization tradition, which implicitly assumes that clients make decisions to seek care, or from the socio-legal perspective, which examines how clients are forced into care. This paper draws from the Network-Episode Model to systematically consider the different social processes through which people come to enter psychiatric treatment by exploring the "stories" told by individuals making their first major contact with the mental health system. We combine the use of qualitative and quantitative methods to examine data from the Indianapolis Network Mental Health Study, a longitudinal study of individuals in treatment at the largest public and voluntary facilities in the city. We analyze detailed self-reports of how they came to use mental health services, classifying these stories as "choice," "coercion," or "muddling through." Using multinomial logit analyses, we examine how factors such as gender, race and diagnosis shape the types of stories that individuals tell. The preliminary results indicate that fewer than half of the stories (45.9%) match the notion of choice underlying the dominant utilization theories. Almost a quarter of respondents (22.9%) report coercion and nearly one-third (31.2%) tell stories that lack a clear agent. Diagnosis and social networks tap differences in how individuals experience entry into care. Individuals diagnosed with bipolar disorder or who have larger, closer social networks are more likely to tell stories of coercion. We discuss the theoretical, methodological, and clinical implications of findings drawn from this examination of clients' storiesmental health utilization coercion

Integrated proteomic and transcriptomic profiling identifies aberrant gene and protein expression in the sarcomere, mitochondrial complex I, and the extracellular matrix in Warmblood horses with myofibrillar myopathy

Abstract Background Myofibrillar myopathy in humans causes protein aggregation, degeneration, and weakness of skeletal muscle. In horses, myofibrillar myopathy is a late-onset disease of unknown origin characterized by poor performance, atrophy, myofibrillar disarray, and desmin aggregation in skeletal muscle. This study evaluated molecular and ultrastructural signatures of myofibrillar myopathy in Warmblood horses through gluteal muscle tandem-mass-tag quantitative proteomics (5 affected, 4 control), mRNA-sequencing (8 affected, 8 control), amalgamated gene ontology analyses, and immunofluorescent and electron microscopy. Results We identified 93/1533 proteins and 47/27,690 genes that were significantly differentially expressed. The top significantly differentially expressed protein CSRP3 and three other differentially expressed proteins, including, PDLIM3, SYNPO2, and SYNPOL2, are integrally involved in Z-disc signaling, gene transcription and subsequently sarcomere integrity. Through immunofluorescent staining, both desmin aggregates and CSRP3 were localized to type 2A fibers. The highest differentially expressed gene CHAC1, whose protein product degrades glutathione, is associated with oxidative stress and apoptosis. Amalgamated transcriptomic and proteomic gene ontology analyses identified 3 enriched cellular locations; the sarcomere (Z-disc & I-band), mitochondrial complex I and the extracellular matrix which corresponded to ultrastructural Z-disc disruption and mitochondrial cristae alterations found with electron microscopy. Conclusions A combined proteomic and transcriptomic analysis highlighted three enriched cellular locations that correspond with MFM ultrastructural pathology in Warmblood horses. Aberrant Z-disc mechano-signaling, impaired Z-disc stability, decreased mitochondrial complex I expression, and a pro-oxidative cellular environment are hypothesized to contribute to the development of myofibrillar myopathy in Warmblood horses. These molecular signatures may provide further insight into diagnostic biomarkers, treatments, and the underlying pathophysiology of MFM

An E321G MYH1 mutation is strongly associated with nonexertional rhabdomyolysis in Quarter Horses.

BackgroundAn E321G mutation in MYH1 was recently identified in Quarter Horses (QH) with immune-mediated myositis (IMM) defined by a phenotype of gross muscle atrophy and myofiber lymphocytic infiltrates.Hypothesis/objectivesWe hypothesized that the MYH1 mutation also was associated with a phenotype of nonexertional rhabdomyolysis. The objective of this study was to determine the prevalence of the MYH1 mutation in QH with exertional (ER) and nonexertional (nonER) rhabdomyolysis.AnimalsQuarter Horses: 72 healthy controls, 85 ER-no atrophy, 56 ER-atrophy, 167 nonER horses selected regardless of muscle atrophy.MethodsClinical and histopathologic information and DNA was obtained from a database for (1) ER &gt; 2 years of age, with or without atrophy and (2) nonER creatine kinase (CK) ≥ 5000 U/L, &lt;5 years of age. Horses were genotyped for E321G MYH1 by pyrosequencing.ResultsThe MYH1 mutation was present in a similar proportion of ER-no atrophy (1/56; 2%) and in a higher proportion of ER-atrophy (25/85; 29%) versus controls (4/72; 5%). The MYH1 mutation was present in a significantly higher proportion of nonER (113/165; 68%) than controls either in the presence (39/42; 93%) or in absence (72/123; 59%) of gross atrophy. Lymphocytes were present in &lt;18% of muscle samples with the MYH1 mutation.Conclusions and clinical importanceAlthough not associated with ER, the MYH1 mutation is associated with atrophy after ER. The MYH1 mutation is highly associated with nonER regardless of whether muscle atrophy or lymphocytic infiltrates are present. Genetic testing will enhance the ability to diagnose MYH1 myopathies (MYHM) in QH

Improving advanced pharmacy practice experiences with an intention/reflection practice

Background and purpose: Experiential learning in pharmacy has the potential to offer transformative experiences for students. Advanced pharmacy practice experiences (APPEs) can be improved if students are encouraged and able to (1) identify and track individual learning gains and interests, and (2) develop self-awareness and intrinsic motivation. Educational activity and setting: The intention/reflection (I/R) practice was developed to address these educational concepts and help facilitate meaningful experiences during APPEs. The I/R tool is a simple, nontechnical, resource-light activity consisting of a set of three to five questions at the beginning, midpoint, and end of an APPE. The questions were designed to help students identify how they can attain meaningful gains from each APPE. Preceptors across three universities designed, implemented, and evaluated the impact of an I/R practice within the context of a variety of APPEs. The APPEs varied between 4, 5, 6, and 10 weeks and were focused on patient care and academia. Findings: Three of the I/R questions were selected for thematic analysis, one at each point of the rotation. These questions were strategically selected because they demonstrate the value resulting from the progressive nature of the I/R tool. The answers to the three questions were descriptively coded to capture the main thought(s) of each student\u27s response. A quarter of students indicated I/R helped them achieve their goals. A retrospective pre-post survey demonstrated statistically significant improvements in all survey items, including (1) ability to identify learning outcomes, (2) focus and motivation, and (3) self-awareness and metacognition

A missense mutation in MYH1 is associated with susceptibility to immune-mediated myositis in Quarter Horses

Abstract Background The cause of immune-mediated myositis (IMM), characterized by recurrent, rapid-onset muscle atrophy in Quarter Horses (QH), is unknown. The histopathologic hallmark of IMM is lymphocytic infiltration of myofibers. The purpose of this study was to identify putative functional variants associated with equine IMM. Methods A genome-wide association (GWA) study was performed on 36 IMM QHs and 54 breed matched unaffected QHs from the same environment using the Equine SNP50 and SNP70 genotyping arrays. Results A mixed model analysis identified nine SNPs within a ~ 2.87 Mb region on chr11 that were significantly (P unadjusted < 1.4 × 10− 6) associated with the IMM phenotype. Associated haplotypes within this region encompassed 38 annotated genes, including four myosin genes (MYH1, MYH2, MYH3, and MYH13). Whole genome sequencing of four IMM and four unaffected QHs identified a single segregating nonsynonymous E321G mutation in MYH1 encoding myosin heavy chain 2X. Genotyping of additional 35 IMM and 22 unaffected QHs confirmed an association (P = 2.9 × 10− 5), and the putative mutation was absent in 175 horses from 21 non-QH breeds. Lymphocytic infiltrates occurred in type 2X myofibers and the proportion of 2X fibers was decreased in the presence of inflammation. Protein modeling and contact/stability analysis identified 14 residues affected by the mutation which significantly decreased stability. Conclusions We conclude that a mutation in MYH1 is highly associated with susceptibility to the IMM phenotype in QH-related breeds. This is the first report of a mutation in MYH1 and the first link between a skeletal muscle myosin mutation and autoimmune disease