Interobserver Agreement and Correlation of an Automated Algorithm for B-Line Identification and Quantification With Expert Sonologist Review in a Handheld Ultrasound Device.

OBJECTIVES: B-lines are ultrasound artifacts that can be used to detect a variety of pathologic lung conditions. Computer-aided methods to detect and quantify B-lines may standardize quantification and improve diagnosis by novice users. We sought to test the performance of an automated algorithm for the detection and quantification of B-lines in a handheld ultrasound device (HHUD). METHODS: Ultrasound images were prospectively collected on adult emergency department patients with dyspnea. Images from the first 124 patients were used for algorithm development. Clips from 80 unique subjects for testing were randomly selected in a predefined proportion of B-lines (0 B-lines, 1-2 B-lines, 3 or more B-lines) and blindly reviewed by five experts using both a manual and reviewer-adjusted process. Intraclass correlation coefficient (ICC) and weighted kappa were used to measure agreement, while an a priori threshold of an ICC (3,k) of 0.75 and precision of 0.3 were used to define adequate performance. RESULTS: ICC between the algorithm and manual count was 0.84 (95% confidence interval [CI] 0.75-0.90), with a precision of 0.15. ICC between the reviewer-adjusted count and the algorithm count was 0.94 (95% CI 0.90-0.96), and the ICC between the manual and reviewer-adjusted counts was 0.94 (95% CI 0.90-0.96). Weighted kappa was 0.72 (95% CI 0.49-0.95), 0.88 (95% CI 0.74-1), and 0.85 (95% CI 0.89-0.96), respectively. CONCLUSIONS: This study demonstrates a high correlation between point-of-care ultrasound experts and an automated algorithm to identify and quantify B-lines using an HHUD. Future research may incorporate this HHUD in clinical studies in multiple settings and users of varying experience levels.

Distinctive requirement of PKCε in the control of Rho GTPases in epithelial and mesenchymally transformed lung cancer cells.

Diacylglycerol (DAG)/phorbol ester-regulated protein kinase C (PKC) isozymes have been widely linked to tumor promotion and the development of a metastatic phenotype. PKCε, an oncogenic member of the PKC family, is abnormally overexpressed in lung cancer and other cancer types. This kinase plays significant roles in proliferation, survival, and migration; however, its role in epithelial-to-mesenchymal transition (EMT) has been scarcely studied. Silencing experiments in non-small lung cancer (NSCLC) cells revealed that PKCε or other DAG-regulated PKCs (PKCα and PKCδ) were dispensable for the acquisition of a mesenchymal phenotype induced by transforming growth factor beta (TGF-ß). Unexpectedly, we found a nearly complete down-regulation of PKCε expression in TGF-ß-mesenchymally transformed NSCLC cells. PMA and AJH-836 (a DAG-mimetic that preferentially activates PKCε) promote ruffle formation in NSCLC cells via Rac1, however they fail to induce these morphological changes in TGF-ß-mesenchymally transformed cells despite their elevated Rac1 activity. Several Rac guanine nucleotide exchange-factors (Rac-GEFs) were also up-regulated in TGF-ß-treated NSCLC cells, including Trio and Tiam2, which were required for cell motility. Lastly, we found that silencing or inhibiting PKCε enhances RhoA activity and stress fiber formation, a phenotype also observed in TGF-ß-transformed cells. Our studies established a distinctive involvement of PKCε in epithelial and mesenchymal NSCLC cells, and identified a complex interplay between PKCε and small GTPases that contributes to regulation of NSCLC cell morphology and motile activity.

Examining differences between P1 versus P2 students as teaching assistants in a P1 skills-based course.

INTRODUCTION: The purpose of this study was to assess differences between the use of first-year (P1; "peer") versus second-year (P2; "near-peer") students as teaching assistants (TA) in a first-year, skills-based course. METHODS: The practicum course assesses competence in the provision of screening services and patient counseling. TAs review weekly material followed by a one-on-one assessment of each student using a grading rubric. Both qualitative and quantitative data were analyzed to determine if there was a difference in performance between the peer and near-peer teaching assistants. RESULTS: Sixteen peer and 33 near-peer TAs were evaluated by 210 students for six different skill assessments in practicum. There was no significant difference between peer and near-peer TAs in both student perception of TA performance and in TA grading of student performance. CONCLUSIONS: There is no difference in the use of peer versus near-peer TAs in evaluating first-year pharmacy students in the skills-based course. Using peer TAs over near-peer TAs can be useful when faced with scheduling and other resource conflicts.

Sexually dimorphic gene expression in the lateral eyes of Euphilomedes carcharodonta (Ostracoda, Pancrustacea).

BACKGROUND: The evolution and development of sexual dimorphism illuminates a central question in biology: How do similar genomes produce different phenotypes? In an XX/XO system especially the state of a sexually dimorphic trait is determined by differences in gene expression, as there are no additional genetic loci in either sex. Here, we examine the XX/XO ostracod crustacean species Euphilomedes carcharodonta. This species exhibits radical sexual dimorphism of their lateral eyes, females have only a tiny simple lateral eye while males have elaborate ommatidial eyes. RESULTS: We find that males express three of nine eye-development gene homologs at significantly higher levels during juvenile eye development, compared to females. We also find that most eye-development genes examined are pleiotropic, with high expression levels during embryonic development as well as during juvenile eye development. Later, in adults, we find that phototransduction genes are expressed at higher levels in males than in females, as we might expect when comparing ommatidial to simple eyes. CONCLUSIONS: We show here that expression changes of a handful of developmental genes may underlie the radical difference in a dimorphic character. This work gives an important point of comparison for studying eye evolution and development in the Pancrustacea.