Massive-Scale RNA-Seq Analysis of Non Ribosomal Transcriptome in Human Trisomy 21

Hybridization- and tag-based technologies have been successfully used in Down syndrome to identify genes involved in various aspects of the pathogenesis. However, these technologies suffer from several limits and drawbacks and, to date, information about rare, even though relevant, RNA species such as long and small non-coding RNAs, is completely missing. Indeed, none of published works has still described the whole transcriptional landscape of Down syndrome. Although the recent advances in high-throughput RNA sequencing have revealed the complexity of transcriptomes, most of them rely on polyA enrichment protocols, able to detect only a small fraction of total RNA content. On the opposite end, massive-scale RNA sequencing on rRNA-depleted samples allows the survey of the complete set of coding and non-coding RNA species, now emerging as novel contributors to pathogenic mechanisms. Hence, in this work we analysed for the first time the complete transcriptome of human trisomic endothelial progenitor cells to an unprecedented level of resolution and sensitivity by RNA-sequencing. Our analysis allowed us to detect differential expression of even low expressed genes crucial for the pathogenesis, to disclose novel regions of active transcription outside yet annotated loci, and to investigate a plethora of non-polyadenilated long as well as short non coding RNAs. Novel splice isoforms for a large subset of crucial genes, and novel extended untranslated regions for known genes—possibly novel miRNA targets or regulatory sites for gene transcription—were also identified in this study. Coupling the rRNA depletion of samples, followed by high-throughput RNA-sequencing, to the easy availability of these cells renders this approach very feasible for transcriptome studies, offering the possibility of investigating in-depth blood-related pathological features of Down syndrome, as well as other genetic disorders

Detection of Babesia bovis and Babesia bigemina in Water Buffaloes (Bubalus bubalis) in Endemic Areas of São Paulo State, Brazil

Abstract Babesiosis is a tick-transmitted disease that causes severe economic losses to the cattle industry in Brazil. Water buffaloes (Bubalus bubalis) are often carriers of Babesia spp., but there are no studies that provide an accurate estimation of this infection in animals raised in regions of endemic stability. This study was conducted to investigate Babesia bovis and B. bigemina infections in 108 water buffaloes (50 calves and 58 adult females) located in areas of São Paulo state, where the animals were continuously exposed to Rhipicephalus microplus ticks. B. bovis and B. bigemina infections were screened by microscopic examination of blood smears, nested PCR (nPCR) and quantitative real-time PCR (qPCR), which were also used to estimate the number of copies (NC) of the cytochrome b (mt-cytB) gene in the blood samples. B. bigemina was found in blood smears of three calves from Alambari herd (all with less than 0.1% parasitemia). Molecular techniques were more sensitive than blood smears to diagnose piroplasms in water buffaloes: 20.37% and 100.00% for B. bovis-infected animals and 59.26% and 100.00% for B. bigemina-infected animals, respectively for nPCR and qPCR. The NC of mt-cytB gene of B. bovis and B. bigemina in blood samples re-* Corresponding author. T. A. Néo et al. 76 vealed significant effects (p < 0.05) of herd-age, species and their interaction. The NC values were higher (p ≤ 0.05) for B. bigemina (2.80 ± 0.06) than for B. bovis (2.61 ± 0.05). Within each herd-age, differences between the species' NC values were found only in Alambari calves, which showed significantly higher (p ≤ 0.05) NC of B. bigemina (3.48 ± 0.13). The calves and cows from Ibaté showed the lowest NC of B. bigemina (2.29 ± 0.13 and 2.63 ± 0.14) and B. bovis (2.54 ± 0.11 and 2.37 ± 0.12), respectively. These data suggest a high prevalence of B. bovis and B. bigemina infection in the buffalo population in endemic areas of São Paulo state

Recombinant Coagulation Factor VIIa in Major Liver Resection

Background: Prevention of bleeding episodes in noncirrhotic patients undergoing partial hepatectomy remains unsatisfactory in spite of improved surgical techniques. The authors conducted a randomized, placebo-controlled, double-blind trial to evaluate the hemostatic effect and safety of recombinant factor VIIa (rFVIIa) in major partial hepatectomy. Methods: Two hundred four noncirrhotic patients were equally randomized to receive either 20 or 80 g/kg rFVIIa or placebo. Partial hepatectomy was performed according to local practice at the participating centers. Patients were monitored for 7 days after surgery. Key efficacy parameters were perioperative erythrocyte requirements (using hematocrit as the transfusion trigger) and blood loss. Safety assessments included monitoring of coagulation-related parameters and Doppler examination of hepatic vessels and lower extremities. Results: The proportion of patients who required perioperative red blood cell transfusion (the primary endpoint) was 37% (23 of 63) in the placebo group, 41% (26of 63) in the 20-g/kg group, and 25% (15 of 59) in the 80-g/kg dose group (logistic regression model; P ‫؍‬ 0.09). Mean erythrocyte requirements for patients receiving erythrocytes were 1,024 ml with placebo, 1,354 ml with 20 g/kg rFVIIa, and 1,036 ml with 80 g/kg rFVIIa (P ‫؍‬ 0.78). Mean intraoperative blood loss was 1,422 ml with placebo, 1,372 ml with 20 g/kg rFVIIa, and 1,073 ml wit