Standardization of islet isolation outcome – A new automatic system to determine pancreatic islet viability

Pancreatic islet transplantation is emerging as a therapeutic approach for patients affected by diabetes. This approach consists of a minimally invasive procedure replacing insulin-producing cells (pancreatic islets). The technique has been proven successful, but limitations have been identified. One of the major challenges of the procedure is the counting of the isolated pancreatic islets, which is currently jeopardized by subjectivity and inaccuracy. Determination of the accurate islet number is a crucial factor in determining the correlation between the isolation product and clinical outcome. In the proposed study, we have developed software capable of objectively evaluating islet numbers and other viability variables by image analysis. This software is based on image processing and feature extraction algorithms for recognition of the area of interest. This is the first step toward standardization of the isolation outcome and potential clinical success predictability.     

Legumes seeds protein isolates in the production of ascorbic acid microparticles

This study evaluated the use of pea (Pisum sativum) and cowpea (Vigna unguiculata) protein isolates (PPI and CPI) in the microencapsulation of ascorbic acid (AA), by spray drying. PPI and CPI were used pure or mixed with maltodextrin (M) as wall materials of AA. The process yield was improved from 66% to 86% by the addition of M (CPI/M and PPI/M); also, AA particles yield was 79%. The AA retention from the drying process was higher in PPI/M (69%) than in CPI/M (66%), and this was attributed to PPI proteolysis profile, demonstrated by the SDS–PAGE electrophoresis. Scanning electron microscopy revealed that AA particles were similarly spherical and shriveled. Mean particle sizes of PPI/M/AA and CPI/M/AA were 7 and 8 μm, respectively. This study showed the exploration of protein isolates from legumes in the development of a microparticulate system containing AA with great potential for use in human health.     

Investigation of metallic nanoparticles adsorbed on the QCM sensor by SEM and AFM techniques

Quartz crystal microbalance (QCM) is known as a very sensitive device used for determination of mass quantity adsorbed on sensor surface. Its detection limits are in the range of ng cm\(^{-2}\). The adsorption mechanism of metallic nanoparticles on QCM sensor was investigated by scanning electron microscopy (SEM) and atomic force microscopy (AFM). This study aims to highlight the importance of QCM applications in nanoparticles deposition field. The layers formed through adsorption process, induced by the oscillations of the QCM sensor, were investigated by AFM for surface topography and for particle mean size values. The morphology of layers and nanoparticles dimensions were determined by SEM. For a more complex investigation of the nanoparticles adsorption mechanism, the chemical composition of layers was achieved using SEM coupled with energy dispersive X-ray spectrometer (SEM-EDS). This preliminary research involved a new approach in characterization of metallic nanoparticles layers to achieve functional assembled monolayers.     

Hyaluronic acid hydrogels incorporating platelet lysate enhance human pulp cell proliferation and differentiation

The restoration of dentine-pulp complex remains a challenge for dentists; nonetheless, it has been poorly addressed. An ideal system should modulate the host response, as well as enable the recruitment, proliferation and differentiation of relevant progenitor cells. Herein was proposed a photocrosslinkable hydrogel system based on hyaluronic acid (HA) and platelet lysate (PL). PL is a cocktail of growth factors (GFs) and cytokines involved in wound healing orchestration, obtained by the cryogenic processing of platelet concentrates, and was expected to provide the HA hydrogels specific biochemical cues to enhance pulp cells’ recruitment, proliferation and differentiation. Stable HA hydrogels incorporating PL (HAPL) were prepared after photocrosslinking of methacrylated HA (Met-HA) previously dissolved in PL, triggered by the Ultra Violet activated photoinitiator Irgacure 2959. Both the HAPL and plain HA hydrogels were shown to be able to recruit cells from a cell monolayer of human dental pulp stem cells (hDPSCs) isolated from permanent teeth. The hDPCs were also seeded directly over the hydrogels (5?×?10⁴ cells/hydrogel) and cultured in osteogenic conditions. Cell metabolism and DNA quantification were higher, in all time-points, for PL supplemented hydrogels (p?<?0,05). Alkaline phosphatase (ALPL) activity and calcium quantification peaks were observed for the HAPL group at 21 days (p?<?0,05). The gene expression for ALPL and COLIA1 was up-regulated at 21 days to HAPL, compared with HA group (p?<?0,05). Within the same time point, the gene expression for RUNX2 did not differ between the groups. Overall, data demonstrated that the HA hydrogels incorporating PL increased the cellular metabolism and stimulate the mineralized matrix deposition by hDPSCs, providing clear evidence of the potential of the proposed system for the repair of damaged pulp/dentin tissue and endodontics regeneration.     

Modeling ozone and aerosol formation and transport in the pacific northwest with the community Multi-Scale Air Quality (CMAQ) modeling system

The Community Multi-Scale Air Quality (CMAQ) modeling system was used to investigate ozone and aerosol concentrations in the Pacific Northwest (PNW) during hot summertime conditions during July 1-15, 1996. Two emission inventories (El) were developed: emissions for the first El were based upon the National Emission Trend 1996 (NET96) database and the BEIS2 biogenic emission model, and emissions for the second El were developed through a "bottom up" approach that included biogenic emissions obtained from the GLOBEIS model. The two simulations showed that elevated PM2.5 concentrations occurred near and downwind of the Interstate-5 corridor along the foothills of the Cascade Mountains and in forested areas of central Idaho. The relative contributions of organic and inorganic aerosols varied by region, but generally organic aerosols constituted the largest fraction of PM2.5. In wilderness areas near the 1-5 corridor, organic carbon from anthropogenic sources contributed approximately 50% of the total organic carbon with the remainder from biogenic precursors, while in wilderness areas in Idaho, biogenic organic carbon accounted for 80% of the total organic aerosol. Regional analysis of the secondary organic aerosol formation in the Columbia River Gorge, Central Idaho, and the Olympics/Puget Sound showed that the production rate of secondary organic carbon depends on local terpene concentrations and the local oxidizing capacity of the atmosphere, which was strongly influenced by anthropogenic emissions. Comparison with observations from 12 IMPROVE sites and 21 ozone monitoring sites showed that results from the two El simulations generally bracketed the average observed PM parameters and that errors calculated for the model results were within acceptable bounds. Analysis across all statistical parameters indicated that the NW-AIRQUEST El solution performed better at predicting PM2.5, PM1, and beta(ext) even though organic carbon PM was over-predicted, and the NET96 El solution performed better with regard to the inorganic aerosols. For the NW-AIRQUEST El solution, the normalized bias was 30% and the normalized absolute error was 49% for PM2.5 mass. The NW-AIRQUEST solution slightly overestimated peak hourly ozone downwind of urban areas, while the NET96 solution slightly underestimated peak values, and both solutions over-predicted average 03 concentrations across the domain by approximately 6 ppb.     

Transferable Antibiotic Resistances in Marketed Edible Grasshoppers (Locusta migratoria migratorioides)

Grasshoppers are the most commonly eaten insects by humans worldwide, as they are rich in proteins and micronutrients. This study aimed to assess the occurrence of transferable antibiotic resistance genes in commercialized edible grasshoppers. To this end, the prevalence of 12 selected genes [aac(6’)‐Ie aph(2″)‐Ia, blaZ, erm(A), erm(B), erm(C), mecA, tet(M), tet(O), tet(S), tet(K), vanA, vanB] coding for resistance to antibiotics conventionally used in clinical practice was determined. The majority of samples were positive for tet(M) (70.0%), tet(K) (83.3%) and blaZ (83.3%). A low percentage of samples were positive for erm(B) (16.7%), erm(C) (26.7%), and aac(6’)‐Ie aph(2″)‐Ia (13.3%), whereas no samples were positive for erm(A), vanA, vanB, tet(O), and mecA. Cluster analysis identified 4 main clusters, allowing a separation of samples on the basis of their country of origin.     

The Occurrence of Beer Spoilage Lactic Acid Bacteria in Craft Beer Production

Beer is one of the world's most ancient and widely consumed fermented alcoholic beverages produced with water, malted cereal grains (generally barley and wheat), hops, and yeast. Beer is considered an unfavorable substrate of growth for many microorganisms, however, there are a limited number of bacteria and yeasts, which are capable of growth and may spoil beer especially if it is not pasteurized or sterile‐filtered as craft beer. The aim of this research study was to track beer spoilage lactic acid bacteria (LAB) inside a brewery and during the craft beer production process. To that end, indoor air and work surface samples, collected in the brewery under study, together with commercial active dry yeasts, exhausted yeasts, yeast pellet (obtained after mature beer centrifugation), and spoiled beers were analyzed through culture‐dependent methods and PCR‐DGGE in order to identify the contaminant LAB species and the source of contamination. Lactobacillus brevis was detected in a spoiled beer and in a commercial active dry yeast. Other LAB species and bacteria ascribed to Staphylococcus sp., Enterobaceriaceae, and Acetobacter sp. were found in the brewery. In conclusion, the PCR‐DGGE technique coupled with the culture‐dependent method was found to be a useful tool for identifying the beer spoilage bacteria and the source of contamination. The analyses carried out on raw materials, by‐products, final products, and the brewery were useful for implementing a sanitization plan to be adopted in the production plant.