SummaryROCK-Myosin II drives fast rounded-amoeboid migration in cancer cells during metastatic dissemination. Analysis of human melanoma biopsies revealed that amoeboid melanoma cells with high Myosin II activity are predominant in the invasive fronts of primary tumors in proximity to CD206+CD163+ tumor-associated macrophages and vessels. Proteomic analysis shows that ROCK-Myosin II activity in amoeboid cancer cells controls an immunomodulatory secretome, enabling the recruitment of monocytes and their differentiation into tumor-promoting macrophages. Both amoeboid cancer cells and their associated macrophages support an abnormal vasculature, which ultimately facilitates tumor progression. Mechanistically, amoeboid cancer cells perpetuate their behavior via ROCK-Myosin II-driven IL-1α secretion and NF-κB activation. Using an array of tumor models, we show that high Myosin II activity in tumor cells reprograms the innate immune microenvironment to support tumor growth. We describe an unexpected role for Myosin II dynamics in cancer cells controlling myeloid function via secreted factors.

There is ongoing debate on whether screening for nonalcoholic fatty liver disease (NAFLD) is worthwhile in high-risk groups. Because of shared risk factors, NAFLD is highly prevalent in patients with coronary artery disease. We aimed to test the hypothesis that NAFLD screening in patients requiring coronary angiogram would identify high-risk patients and predict long-term clinical outcomes. This was a prospective cohort study. NAFLD screening was performed by abdominal ultrasonography before coronary angiogram in 612 consecutive patients. At baseline, 356 (58.2%) patients had NAFLD. NAFLD patients, compared with those without, were more likely to have >50% stenosis in one or more coronary arteries (84.6% vs. 64.1%; P < 0.001) and therefore require percutaneous coronary intervention (68.3% vs. 43.4%; P < 0.001). During 3,679 patient-years of follow-up, 47 (13.2%) NAFLD patients and 59 (23.0%) patients without NAFLD died (age-and sex-adjusted hazard ratio [aHR]: 0.36; 95% confidence interval [CI]: 0.18-0.70; P 5 0.003). Composite cardiovascular outcomes (cardiovascular deaths, nonfatal myocardial infarction, heart failure, or secondary interventions) were similar between groups (36.5% vs. 37.1%; aHR, 0.90; 95% CI: 0.69-1.18). Older age and diabetes were the only independent factors associated with cardiovascular events. Only 2 patients, both in the NAFLD group, died of primary liver cancer. No other patients developed liver-related complications. Conclusion: In patients with clinical indications for coronary angiogram, the presence of NAFLD is associated with coronary artery stenosis and need for coronary intervention, but not increased mortality or cardiovascular complications. Liver cancer and cirrhotic complications are rare. Our data do not support NAFLD screening in this patient group at present, but studies with a longer duration of follow-up are needed. (HEPATOLOGY 2016;63:754-763)

Based on findings in small RNA-sequencing (Seq) data analysis, we developed highly sensitive and specific real-time reverse transcription (RT)-PCR assays with locked nucleic acid probes targeting the abundantly expressed leader sequences of Middle East respiratory syndrome coronavirus (MERS-CoV) and other human coronaviruses. Analytical and clinical evaluations showed their noninferiority to a commercial multiplex PCR test for the detection of these coronaviruses. C oronaviruses (CoVs) have repeatedly crossed species barriers, and some have emerged as important human pathogens (1, 2). Human coronavirus (HCoV)-229E, HCoV-OC43, HCoV-NL63, and HCoV-HKU1 predominantly cause mild upper respiratory tract infections, while severe acute respiratory syndrome CoV (SARS-CoV) and Middle East respiratory syndrome CoV (MERS-CoV) frequently cause severe pneumonia with extrapulmonary manifestations (3-6). Highly sensitive and specific laboratory diagnostic tests are essential for the control of emerging CoV outbreaks (7). The gold standard for the laboratory diagnosis of CoV infection is the isolation of infectious virus from the respiratory tract and/or other clinical specimens. However, most CoVs are either difficult or dangerous to culture in cell lines (8, 9). The need for convalescent-phase samples and potential false-positive results due to cross-reactivity with other CoVs limit the use of serum antibody detection assays in the acute setting (10). The overall sensitivity of antigen detection assays is inferior to that of molecular assays such as reverse transcription (RT)-PCR (11,12). With the increasing availability of molecular diagnostic facilities and expertise in clinical microbiology laboratories worldwide, RT-PCR has become the test of choice for diagnosing CoV infections (7,(13)(14)(15).Traditionally, the preferred targets of RT-PCR assays are genes that are conserved and/or abundantly expressed from the viral genome (16). For CoVs, the most commonly employed targets include the structural nucleocapsid (N) and spike (S) genes, and the nonstructural RNA-dependent RNA polymerase (RdRp) and replicase ORF1a/b genes (4, 7). Recently, other unique noncoding genome regions not present in related CoVs have also been utilized to develop an RT-PCR for the emerging MERS-CoV (7, 13-15). The World Health Organization (WHO) recommends using the upE assay (regions upstream of the envelope [E] gene) for laboratory screening of suspected MERS cases, followed by confirmation with either the ORF1a or ORF1b assay (7). Notably, a number of single nucleotide mismatches at different positions included in the upE assay forward primer and probe have been detected in recent strains of MERS-CoV and may affect the sensitivity of this assay (17). We hypothesize that additional gene targets may be suitable for design of RT-PCR assays for CoVs and would increase the options of molecular diagnosis for circulating and emerging CoV infections. In this study, we designed and evaluated novel real-time RT-PCR assays with locked nucleic acid (LNA) pro...

No abstract

No abstract