CUT-PCR: CRISPR-mediated, ultrasensitive detection of target DNA using PCR

Circulating tumor DNA (ctDNA) has emerged as a tumor-specific biomarker for the early detection of various cancers. To date, several techniques have been devised to enrich the extremely small amounts of ctDNA present in plasma, but they are still insufficient for cancer diagnosis, especially at the early stage. Here, we developed a novel method, CUT (CRISPR-mediated, Ultrasensitive detection of Target DNA)-PCR, which uses CRISPR endonucleases to enrich and detect the extremely small amounts of tumor DNA fragments among the much more abundant wild-type DNA fragments by specifically eliminating the wild-type sequences. We computed that by using various orthologonal CRISPR endonucleases such as SpCas9 and FnCpf1, the CUT-PCR method would be applicable to 80% of known cancer-linked substitution mutations registered in the COSMIC database. We further verified that CUT-PCR together with targeted deep sequencing enables detection of a broad range of oncogenes with high sensitivity (<0.01%) and accuracy, which is superior to conventional targeted deep sequencing. In the end, we successfully applied CUT-PCR to detect sequences with oncogenic mutations in the ctDNA of colorectal cancer patients' blood, suggesting that our technique could be adopted for diagnosing various types of cancer at early stages

Fasciola hepatica IN BOVINES IN BRAZIL: DATA AVAILABILITY AND SPATIAL DISTRIBUTION

Fasciolosis is a disease of importance for both veterinary and public health. For the first time, georeferenced prevalence data of Fasciola hepatica in bovines were collected and mapped for the Brazilian territory and data availability was discussed. Bovine fasciolosis in Brazil is monitored on a Federal, State and Municipal level, and to improve monitoring it is essential to combine the data collected on these three levels into one dataset. Data were collected for 1032 municipalities where livers were condemned by the Federal Inspection Service (MAPA/SIF) because of the presence of F. hepatica. The information was distributed over 11 states: Espírito Santo, Goiás, Minas Gerais, Mato Grosso do Sul, Mato Grosso, Pará, Paraná, Rio de Janeiro, Rio Grande do Sul, Santa Catarina and São Paulo. The highest prevalence of fasciolosis was observed in the southern states, with disease clusters along the coast of Paraná and Santa Catarina and in Rio Grande do Sul. Also, temporal variation of the prevalence was observed. The observed prevalence and the kriged prevalence maps presented in this paper can assist both animal and human health workers in estimating the risk of infection in their state or municipality

Tropical Fruit Pulps: Processing, Product Standardization and Main Control Parameters for Quality Assurance

ABSTRACT Fruit pulp is the most basic food product obtained from fresh fruit processing. Fruit pulps can be cold stored for long periods of time, but they also can be used to fabricate juices, ice creams, sweets, jellies and yogurts. The exploitation of tropical fruits has leveraged the entire Brazilian fruit pulp sector due mainly to the high acceptance of their organoleptic properties and remarkable nutritional facts. However, several works published in the last decades have pointed out unfavorable conditions regarding the consumption of tropical fruit pulps. This negative scenario has been associated with unsatisfactory physico-chemical and microbiological parameters of fruits pulps as outcomes of little knowledge and improper management within the fruit pulp industry. There are protocols for delineating specific identity and quality standards (IQSs) and standardized good manufacturing practices (GMP) for fruit pulps, which also embrace standard operating procedures (SOPs) and hazard analysis and critical control points (HACCP), although this latter is not considered mandatory by the Brazilian legislation. Unfortunately, the lack of skilled labor, along with failures in complying established protocols have impaired quality of fruit pulps. It has been necessary to collect all information available with the aim to identify the most important hazards within fruit pulp processing lines. Standardizing methods and practices within the Brazilian fruit pulp industry would assurance high quality status to tropical fruit pulps and the commercial growth of this vegetal product towards international markets