One-Step Preservation of Phosphoproteins and Tissue Morphology at Room Temperature for Diagnostic and Research Specimens

BACKGROUND: There is an urgent need to measure phosphorylated cell signaling proteins in cancer tissue for the individualization of molecular targeted kinase inhibitor therapy. However, phosphoproteins fluctuate rapidly following tissue procurement. Snap-freezing preserves phosphoproteins, but is unavailable in most clinics and compromises diagnostic morphology. Formalin fixation preserves tissue histomorphology, but penetrates tissue slowly, and is unsuitable for stabilizing phosphoproteins. We originated and evaluated a novel one-step biomarker and histology preservative (BHP) chemistry that stabilizes signaling protein phosphorylation and retains formalin-like tissue histomorphology with equivalent immunohistochemistry in a single paraffin block. RESULTS: Total protein yield extracted from BHP-fixed, routine paraffin-embedded mouse liver was 100% compared to snap-frozen tissue. The abundance of 14 phosphorylated proteins was found to be stable over extended fixation times in BHP fixed paraffin embedded human colon mucosa. Compared to matched snap-frozen tissue, 8 phosphoproteins were equally preserved in mouse liver, while AMPKβ1 Ser108 was slightly elevated after BHP fixation. More than 25 tissues from mouse, cat and human specimens were evaluated for preservation of histomorphology. Selected tissues were evaluated in a multi-site, independent pathology review. Tissue fixed with BHP showed equivalent preservation of cytoplasmic and membrane cytomorphology, with significantly better nuclear chromatin preservation by BHP compared to formalin. Immunohistochemical staining of 13 non-phosphorylated proteins, including estrogen receptor alpha, progesterone receptor, Ki-67 and Her2, was equal to or stronger in BHP compared to formalin. BHP demonstrated significantly improved immunohistochemical detection of phosphorylated proteins ERK Thr202/Tyr204, GSK3-α/β Ser21/Ser9, p38-MAPK Thr180/Tyr182, eIF4G Ser1108 and Acetyl-CoA Carboxylase Ser79. CONCLUSION: In a single paraffin block BHP preserved the phosphorylation state of several signaling proteins at a level comparable to snap-freezing, while maintaining the full diagnostic immunohistochemical and histomorphologic detail of formalin fixation. This new tissue fixative has the potential to greatly facilitate personalized medicine, biobanking, and phospho-proteomic research

Don\u27t Just Like Me, Promote Me: How Attachment and Attitude Influence Brand Related Behaviors on Social Media

As social media marketing becomes more pervasive, questions continue to emerge regarding utilizing the medium strategically to maximize positive brand-related outcomes. Marketers are increasingly seeking guidance for targeting consumers who will interact and behave in ways that are meaningful to the brand on social media. Understanding how individual differences among consumers can influence social media behaviors linked to valuable organizational outcomes is crucial for managers seeking to justify social media marketing expenditures. This research addresses that issue by applying Attachment Theory to social media. In Study 1, we examine the roles of two individual difference factors, attitude toward social media and attachment to social media (ASM), in predicting token and meaningful behaviors on social media. We find that while attitude toward social media and ASM are both related to token behaviors, only ASM predicts meaningful behaviors. In Study 2, we investigate attachment and attitude toward the brand and social media as predictors of offline and social media brand advocacy and demonstrate that individuals who are attached to or have a positive attitude toward the brand are more likely to engage in offline advocacy. Further, ASM adds incremental explanatory power, beyond attitude and attachment to the brand, in predicting advocacy via social media. In Study 3, we examine likelihood of advocating for a brand via social media as a moderator and find that even consumers who are not likely to be offline advocates are more likely to advocate for the brand on social media if they are strongly attached to social media. In sum, the results indicate that ASM is an important predictor of meaningful social media behaviors and is a new means by which marketers can identify consumers who are more likely to perform meaningful behaviors for brands via social media

Neural Mechanisms Underlying Learning Following Semantic Mediation Treatment in a Case of Phonologic Alexia

Patients with phonologic alexia can be trained to read semantically impoverished words (e.g., functors) by pairing them with phonologically-related semantically rich words (e.g, nouns). What mechanisms underlie success in this cognitive re-training approach? Does the mechanism change if the skill is “overlearned”, i.e., practiced beyond criterion? We utilized fMRI pre- and post-treatment, and after overlearning, to assess treatment-related functional reorganization in a patient with phonologic alexia, two years post left temporoparietal stroke. Pre-treatment, there were no statistically significant differences in activation profiles across the sets of words. Post-treatment, accuracy on the two trained sets improved. Compared with untrained words, reading trained words recruited larger and more significant clusters of activation in the right hemisphere, including right inferior frontal and inferior parietal cortex. Post-overlearning, with near normal performance on overlearned words, predominant activation shifted to left hemisphere regions, including perilesional activation in superior parietal lobe, when reading overlearned vs. untrained words