Identification and characterization of microRNAs expressed in the African malaria vector Anopheles funestus life stages using high throughput sequencing

Background: Over the past several years, thousands of microRNAs (miRNAs) have been identified in the genomes of various insects through cloning and sequencing or even by computational prediction. However, the number of miRNAs identified in anopheline species is low and little is known about their role. The mosquito Anopheles funestus is one of the dominant malaria vectors in Africa, which infects and kills millions of people every year. Therefore, small RNA molecules isolated from the four life stages (eggs, larvae, pupae and unfed adult females) of An. funestus were sequenced using next generation sequencing technology. Results: High throughput sequencing of four replicates in combination with computational analysis identified 107 mature miRNA sequences expressed in the An. funestus mosquito. These include 20 novel miRNAs without sequence identity in any organism and eight miRNAs not previously reported in the Anopheles genus but are known in non-anopheles mosquitoes. Finally, the changes in the expression of miRNAs during the mosquito development were determined and the analysis showed that many miRNAs have stage-specific expression, and are co-transcribed and co-regulated during development. Conclusions: This study presents the first direct experimental evidence of miRNAs in An. funestus and the first profiling study of miRNA associated with the maturation in this mosquito. Overall, the results indicate that miRNAs play important roles during the growth and development. Silencing such molecules in a specific life stage could decrease the vector population and therefore interrupt malaria transmission.IS

Peritoneal carcinomatosis index as a predictor of diaphragmatic involvement in stage III and IV ovarian cancer

Antoni Llueca,1–3 Anna Serra,1–3 José Luis Herraiz,2 Isabel Rivadulla,1,4 Luis Gomez-Quiles,1,4 Juan Gilabert-Estelles,5,6 Javier Escrig1,3,4On behalf of the MUAPOS (Multidisciplinary Unit of Abdominal Pelvic Oncology Surgery) working group 1Multidisciplinary Unit of Abdominal Pelvic Oncology Surgery (MUAPOS), University General Hospital of Castellón, Castellón, Spain; 2Department of Obstetrics and Gynecology, University General Hospital of Castellón, Castellón, Spain; 3Department of Medicine, Universitat Jaume I, Castellón, Spain; 4Department of General Surgery, University General Hospital of Castellón, Castellón, Spain; 5Department of Obstetrics and Gynecology, University General Hospital of Valencia, Valencia, Spain; 6Department of Pediatrics, Obstetrics and Gynecology, University of Valencia, Valencia, Spain Objective: To analyze the surgical outcomes and diaphragmatic involvement in stage III and IV ovarian cancer.Patients and methods: All patients with stage III–IV ovarian cancer between January 2013 and January 2016 were included. The outcomes of interest reviewed were as follows: surgical (complications, mortality), peritoneal carcinomatosis index (PCI), rate of complete resection, and disease-free interval and survival.Results: Fifty-seven patients were included, 38 (67%) with diaphragmatic involvement; in 10 cases (18%), diaphragmatic resection was required. Optimal cytoreduction (OCR) was obtained in 49 cases (86%). The PCI was >10 in 31 cases (54%). Respiratory complications occurred in 10 cases (18%) and mortality in 3 (5%). Disease-free survival rate in 3 years was 53%, being 87% in cases without diaphragmatic involvement. The overall survival rate in 3 years is 46%, 83% in the cases without diaphragmatic involvement and 27% in cases with affectation (p<0.05). In cases of OCR, 3 year survival rate was 65%. In the multivariate analysis for the overall survival of cases with OCR, the only independent prognostic factor found was the operative PCI. A strong correlation was found between the total PCI and the diaphragmatic PCI (p<0.001). With a PCI >10, virtually all cases will present diaphragmatic involvement (p<0.05).Conclusion: The tumor burden is different in stages III and IV of advanced ovarian cancer and the PCI is an effective method to quantify it. The PCI constitutes an independent prognostic factor for the advanced stages of ovarian cancer. A PCI >10 constitutes a useful prognostic factor of the affectation and forces the surgeon to thoroughly review both diaphragms.Keywords: advanced ovarian cancer, peritoneal cancer index, diaphragmatic involvement, upper abdominal surgery, carcinomatosi