The Localization and Function of Novel Tetrahymena thermophila Cytoskeletal Genes BBC29 and BBC39

Ciliary function is known to play an important role in many human conditions, including chronic sinus and pulmonary diseases and problems with infertility. Cilia are cytoskeletal structures that protrude from the cell body to facilitate movement. Ciliary structure is conserved throughout eukaryotes, from unicellular to multicellular organisms, including humans. A clear understanding the proteins that compose cilia and how they interact with one another will increase our knowledge about important cytoskeletal elements. Because cilia are difficult to study in multicellular organisms, the unicellular eukaryote Tetrahymena thermophila serves as a useful model for the study of cytoskeletal genes, due to their robust and longitudinally organized cilia. Here, we examine two novel kinetodesmal fiber proteins in T. thermophila, BBC29 and BBC39. Kinetodesmal fibers stabilize and organize cytoskeletal elements to ensure that all the cilia beat together and in the same orientation to maximize the effects of their beating. Genes BBC29 and BBC39 were isolated using PCR amplification, cloned into pICY-gtw vectors, and transformed into T. thermophila. The pICY vector included an inducible 3’ MTT cadmium promoter and a 5’ yellow fluorescent protein tag to allow visualization of the proteins’ location in live cells under UV light. Both genes localized to the kinetodesmal fiber structures, indicating their role to help stabilize and coordinate ciliary movement. We examined the interaction between the two proteins using co-immunoprecipitation to determine if the two proteins are dependent on each other for proper function. Our results demonstrate the complex interactions that occur between cytoskeletal proteins, such as those in the kinetodesmal fibers of cilia, and illuminate the need to further understand the relationship among the many proteins that allow for proper ciliary function. Because cytoskeletal proteins are often highly conserved between species, results gleaned through study with T. thermophila may be extended to elucidate the biochemical basis of human ciliary diseases

Transcriptome profiling of the rice blast fungus during invasive plant infection and in vitro stresses

<p>Abstract</p> <p>Background</p> <p>Rice blast is the most threatening disease to cultivated rice. <it>Magnaporthe oryzae</it>, its causal agent, is likely to encounter environmental challenges during invasive growth in its host plants that require shifts in gene expression to establish a compatible interaction. Here, we tested the hypothesis that gene expression patterns during <it>in planta </it>invasive growth are similar to <it>in vitro </it>stress conditions, such as nutrient limitation, temperature up shift and oxidative stress, and determined which condition most closely mimicked that of <it>in planta </it>invasive growth. Gene expression data were collected from these <it>in vitro </it>experiments and compared to fungal gene expression during the invasive growth phase at 72 hours post-inoculation in compatible interactions on two grass hosts, rice and barley.</p> <p>Results</p> <p>We identified 4,973 genes that were differentially expressed in at least one of the <it>in planta </it>and <it>in vitro </it>stress conditions when compared to fungal mycelia grown in complete medium, which was used as reference. From those genes, 1,909 showed similar expression patterns between at least one of the <it>in vitro </it>stresses and rice and/or barley. Hierarchical clustering of these 1,909 genes showed three major clusters in which <it>in planta </it>conditions closely grouped with the nutrient starvation conditions. Out of these 1,909 genes, 55 genes and 129 genes were induced and repressed in all treatments, respectively. Functional categorization of the 55 induced genes revealed that most were either related to carbon metabolism, membrane proteins, or were involved in oxidoreduction reactions. The 129 repressed genes showed putative roles in vesicle trafficking, signal transduction, nitrogen metabolism, or molecular transport.</p> <p>Conclusions</p> <p>These findings suggest that <it>M. oryzae </it>is likely primarily coping with nutrient-limited environments at the invasive growth stage 72 hours post-inoculation, and not with oxidative or temperature stresses.</p

Genome-Wide Expression Profiling of Five Mouse Models Identifies Similarities and Differences with Human Psoriasis

Development of a suitable mouse model would facilitate the investigation of pathomechanisms underlying human psoriasis and would also assist in development of therapeutic treatments

Implementation of a validated post-operative opioid nomogram into clinical gynecologic surgery practice: A quality improvement initiative

Objectives: The Gynecologic Oncology Postoperative Opioid use Predictive (GO-POP) calculator is a validated tool to provide evidence-based guidance on post-operative opioid prescribing. The objective of this study was to evaluate the impact of the implementation of GO-POP within an academic Gynecologic Oncology division. Methods: Two cohorts of patients (pre-implementation and post-implementation) who underwent surgery were compared with reference to GO-POP calculator implementation. All patients were included in the post-implementation group, regardless of GO-POP calculator use. An additional expanded-implementation cohort was used to compare pain control between GO-POP users and non-GO-POP users prospectively. Wilcoxon rank sum tests or ANOVA for continuous variables and Chi-square or Fisher’s exact tests were used to categorical variables. Results: The median number of pills prescribed post-operatively decreased from 15 pills (Q1: 10, Q3: 20) to 10 pills (Q1: 8, Q3: 14.8) after implementation (p < 0.001). In the expanded-implementation cohort (293 patients), 41% patients were prescribed opioids using the GO-POP calculator. An overall median of 10 pills were prescribed with no difference by GO-POP calculator use (p = 0.26). Within the expanded-implementation cohort, refill requests (5% vs 9.2%; p = 0.26), clinician visits (0.8% vs 0.6%, p = 1), ED or urgent care visits (0% vs 2.3%, p = 0.15) and readmissions (0% vs 1.7%, p = 0.27) for pain did not differ between those prescribed opioids with and without the GO-POP calculator. Conclusions: A 33% reduction in post-operative opioid pills prescribed was seen following implementation of the GO-POP calculator into the Gynecologic Oncology division without increasing post-operative pain metrics or encounters for refill requests