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MicroRNAs (miRNAs) constitute a class of small noncodingRNAs that play important roles in a variety of biological processes including development, apoptosis, proliferation, and differentiation. Here we show that the expression of miR-199a and miR-199a* (miR-199a/a*), which are processed from the same precursor, is confined to fibroblast cells among cultured cell lines. The fibroblast-specific expression pattern correlated well with methylation patterns: gene loci on chromosome 1 and 19 were fully methylated in all examined cell lines but unmethylated in fibroblasts. Transfection of miR-199a and/or -199a* mimetics into several cancer cell lines caused prominent apoptosis with miR-199a* being more pro-apoptotic. The mechanism underlying apoptosis induced by miR-199a was caspasedependent, whereas a caspase-independent pathway was involved in apoptosis induced by miR-199a* in A549 cells. By employing microarray and immunoblotting analyses, we identified the MET proto-oncogene as a target of miR-199a*. Studies with a luciferase reporter fused to the 3-untranslated region of the MET gene demonstrated miR-199a*-mediated down-regulation of luciferase activity through a binding site of miR-199a*. Interestingly, extracellular signal-regulated kinase 2 (ERK2) was also down-regulated by miR-199a*. Coordinated down-regulation of both MET and its downstream effector ERK2 by miR199a* may be effective in inhibiting not only cell proliferation but also motility and invasive capabilities of tumor cells.

Background Cerebral Performance Category (CPC), Modified Rankin Scale (mRS) and discharge disposition are commonly used to determine outcomes following cardiac arrest. This study tested the association between these outcome measures. Methods Retrospective chart review of subjects who survived to hospital discharge between 1/1/2006 and 12/31/2009 was conducted. Charts were reviewed for outcomes (CPC, mRS, and discharge disposition). Discharge disposition was classified in 6 categories: home with no services, home with home healthcare, acute rehabilitation facility, skilled nursing facility, long term acute care facility, and hospice. Intra-and inter-rater reliabilities were calculated for outcome measures. Rates of “good outcome” (defined as a CPC of 1–2, mRS of 0–3, or discharge disposition to home or acute rehabilitation facility) were also determined. Kendall’s tau correlation coefficients explored relationships among measures. Results A total of 211 charts were reviewed. Mean age was 60 years (SD 16), the majority (75%) were white males, in- and out-of hospital cardiac arrests were equally prevalent, and ventricular dysrhythmia was most common (N=109, 52%). Half of the subjects were comatose following resuscitation and 75 (35%) received therapeutic hypothermia. Inter-rater percentage agreement for CPC and mRS abstraction was 95.24% (kappa 0.89, p<0.001) and 95.24% (kappa 0.90, p<0.001) respectively. “Good outcomes” were found in 44 subjects (20%) using the CPC definition, 47 subjects (22%) using the mRS definition, and 129 subjects (61%) subjects using discharge disposition definition. There was fair relationship between the CPC and mRS (tau 0.43) and poor relationships between CPC and discharge disposition (tau 0.23) and between mRS and discharge disposition (tau 0.25). Conclusions Determination of the CPC, mRS and discharge disposition at hospital discharge is reliable from chart review. These instruments provide widely differing estimates of “good outcome.” Agreement between these measures ranges from poor to fair. A more nuanced outcome measure designed for the post-cardiac arrest population is needed.

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by progressive weakness from loss of motor neurons. The fundamental pathogenic mechanisms are unknown and recent evidence is implicating a significant role for abnormal exon splicing and RNA processing. Using new comprehensive genomic technologies, we studied exon splicing directly in 12 sporadic ALS and 10 control lumbar spinal cords acquired by a rapid autopsy system that processed nervous systems specifically for genomic studies. ALS patients had rostral onset and caudally advancing disease and abundant residual motor neurons in this region. We created two RNA pools, one from motor neurons collected by laser capture microdissection and one from the surrounding anterior horns. From each, we isolated RNA, amplified mRNA, profiled whole-genome exon splicing, and applied advanced bioinformatics. We employed rigorous quality control measures at all steps and validated findings by qPCR. In the motor neuron enriched mRNA pool, we found two distinct cohorts of mRNA signals, most of which were up-regulated: 148 differentially expressed genes (P ≤ 10−3) and 411 aberrantly spliced genes (P ≤ 10−5). The aberrantly spliced genes were highly enriched in cell adhesion (P ≤ 10−57), especially cell–matrix as opposed to cell–cell adhesion. Most of the enriching genes encode transmembrane or secreted as opposed to nuclear or cytoplasmic proteins. The differentially expressed genes were not biologically enriched. In the anterior horn enriched mRNA pool, we could not clearly identify mRNA signals or biological enrichment. These findings, perturbed and up-regulated cell–matrix adhesion, suggest possible mechanisms for the contiguously progressive nature of motor neuron degeneration. Data deposition: GeneChip raw data (CEL-files) have been deposited for public access in the Gene Expression Omnibus (GEO), , accession number GSE18920.