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Background: Bkv-miR-B1-5p is a viral micro-RNA (miRNA) specifically produced during BK polyomavirus (BKPyV) replication. Recent studies have suggested using bkv-miR-B1-5p as a biomarker to monitor viral infection and predict complications in kidney transplant patients. To identify the technical limitations of this miRNA quantification in biological samples, knowledge of its stability and distribution in the extracellular compartment is necessary. Moreover, a proof of concept for using bkv-miR-B1-5p as a biomarker of active replication in chronic infection is still missing in the published literature. Methods: The stability of bkv-miR-B1-5p was evaluated in samples derived from cell cultures and in urine from BKPyV-infected kidney transplant recipients. The miRNA was quantified in different fractions of the extracellular compartment, including exosomes, and protein binding was evaluated. Finally, we developed an in vitro model for chronic culture of BKPyV clinical isolates to observe changes in the bkv-miR-B1-5p level during persistent infections. Results: Bkv-miR-B1-5p is a stable biomarker in samples from humans and in vitro experiments. Marginally associated with the exosomes, most of the circulating bkv-miR-B1-5p is bound to proteins, especially Ago2, so the miRNA quantification does not require specific exosome isolation. The bkv-miR-B1-5p level is predictable of viral infectivity, which makes it a potential specific biomarker of active BKPyV replication after kidney transplantation.

In 2000, the Consumer Goods Forum established the Global Food Safety Initiative (GFSI) to increase the safety of the world's food supply and to harmonize food safety regulations worldwide. In 2013, a university research team in conjunction with Diversey Consulting (Sealed Air), the Consumer Goods Forum, and officers of GFSI solicited input from more than 15,000 GFSI-certified food producers worldwide to determine whether GFSI certification had lived up to these expectations. A total of 828 usable questionnaires were analyzed, representing about 2,300 food manufacturing facilities and food suppliers in 21 countries, mainly across Western Europe, Australia, New Zealand, and North America. Nearly 90% of these certified suppliers perceived GFSI as being beneficial for addressing their food safety concerns, and respondents were eight times more likely to repeat the certification process knowing what it entailed. Nearly three-quarters (74%) of these food manufacturers would choose to go through the certification process again even if certification were not required by one of their current retail customers. Important drivers for becoming GFSI certified included continuing to do business with an existing customer, starting to do business with new customer, reducing the number of third-party food safety audits, and continuing improvement of their food safety program. Although 50% or fewer respondents stated that they saw actual increases in sales, customers, suppliers, or employees, significantly more companies agreed than disagreed that there was an increase in these key performance indicators in the year following GFSI certification. A majority of respondents (81%) agreed that there was a substantial investment in staff time since certification, and 50% agreed there was a significant capital investment. This survey is the largest and most representative of global food manufacturers conducted to date.

Confocal laser endomicroscopy (CLE) uses a low-energy laser light source to obtain microscopic histology images of bladder tissue exposed to a fluorescent dye. To evaluate the feasibility of using CLE with two fluorophores: fluorescein (FLUO) and hexylaminolevulinate (HAL) to determine histologic and cytologic bladder cancer criteria.Patients eligible for HAL-photodynamic diagnosis-assisted transurethral resection of bladder tumor were included. The procedures were performed with the patient under regional or general anesthesia (60-90 minutes) after bladder instillation of HAL (50 mL, 8 mmol/L; HexvixTwenty-one patients were included, 12 examined by CVI and 9 by CVII. Sample examination on CVI after HAL-CLE-only histologic analysis was not possible because HAL is mostly cytoplasmic and gives poor details on cellular architecture. On the contrary, FLUO-CLE gives good extracellular architecture and not clear information of nucleocytoplasmic abnormality. Samples on CVII for seven out of nine patients clearly showed cytoplasm of suspect cells and nuclei. In real time, fluorescence observed on bandwidth (673-800 nm) with HAL and FLUO was associated with the presence of cancer, with a sensibility and specificity of 80% and 100%, respectively.Real-time cytodetection was feasible using two fluorophores (FLUO and HAL) and the new system of CVII. This technology was useful to observe cytoplasm, nuclei, and nucleocytoplasmic abnormality, but an improved system is necessary (to overcome the overlapping of fluorescence) to increase the specificity.