Visualization and quantitation of the expression of microRNAs and their target genes in neuroblastoma single cells using imaging cytometry

<p>Abstract</p> <p>Background</p> <p>MicroRNAs (miRNAs) are regulatory molecules that play an important role in many physiological processes, including cell growth, differentiation, and apoptosis. In addition to modulating normal cellular functions, it has also been reported that miRNAs are involved in the development of many pathologies, including cardiovascular diseases, cancer, inflammation, and neurodegeneration. Methods for the sensitive detection and measurement of specific miRNAs and their cellular targets are essential for both basic research endeavours, as well as diagnostic efforts aimed at understanding the role of miRNAs in disease processes.</p> <p>Findings</p> <p>In this study, we describe a novel, imaging cytometry-based protocol that allows for simultaneous visualisation and quantification of miRNAs and their putative targets. We validated this methodology in a neuronal cell line by examining the relationship of the miRNA miR-124 and its known target, cyclin dependent kinase 6 (CDK6). We found that ectopic overexpression of miR-124 resulted in the downregulation of CDK6, decreased cellular proliferation, and induced cellular morphological changes.</p> <p>Conclusions</p> <p>This method is suitable for analysing the expression and cellular localisation of miRNAs and target proteins in small cell subsets within a heterogeneous cell suspension. We believe that our cytometry-based methodology will be easily adaptable to miRNA studies in many areas of biomedical research including neuroscience, stem cell biology, immunology, and oncology.</p

Anti-inflammatory Components from Functional Foods for Obesity

Obesity, defined as excessive fat accumulation that may impair health, has been described throughout human history, but it has now reached epidemic proportions with the WHO estimating that 39% of the world’s adults over 18 years of age were overweight or obese in 2016. Obesity is a chronic low-grade inflammatory state leading to organ damage with an increased risk of common diseases including cardiovascular and metabolic disease, non-alcoholic fatty liver disease, osteo-arthritis and some cancers. This inflammatory state may be influenced by adipose tissue hypoxia and changes in the gut microbiota. There has been an increasing focus on functional foods and nutraceuticals as treatment options for obesity as drug treatments are limited in efficacy. This chapter summarises the importance of anthocyanin-containing fruits and vegetables, coffee and its components, tropical fruit and food waste as sources of phytochemicals for obesity treatment. We emphasise that preclinical studies can form the basis for clinical trials to determine the effectiveness of these treatments in humans

Interaction of wind and cold‐season hydrologic processes on erosion from complex topography following wildfire in sagebrush steppe

Wildfire is a natural component of sagebrush (Artemisia spp.) steppe rangelands that induces temporal shifts in plant community physiognomy, ground surface conditions, and erosion rates. Fire alteration of the vegetation structure and ground cover in these ecosystems commonly amplifies soil losses by wind- and water-driven erosion. Much of the fire-related erosion research for sagebrush steppe has focused on either erosion by wind over gentle terrain or water-driven erosion under high-intensity rainfall on complex topography. However, many sagebrush rangelands are geographically positioned in snow-dominated uplands with complex terrain in which runoff and sediment delivery occur primarily in winter months associated with cold-season hydrology. Current understanding is limited regarding fire effects on the interaction of wind- and cold-season hydrologic-driven erosion processes for these ecosystems. In this study, we evaluated fire impacts on vegetation, ground cover, soils, and erosion across spatial scales at a snow-dominated mountainous sagebrush site over a 2-year period post-fire. Vegetation, ground cover, and soil conditions were assessed at various plot scales (8 m2 to 3.42 ha) through standard field measures. Erosion was quantified through a network of silt fences (n = 24) spanning hillslope and side channel or swale areas, ranging from 0.003 to 3.42 ha in size. Sediment delivery at the watershed scale (129 ha) was assessed by suspended sediment samples of streamflow through a drop-box v-notch weir. Wildfire consumed nearly all above-ground live vegetation at the site and resulted in more than 60% bare ground (bare soil, ash, and rock) in the immediate post-fire period. Widespread wind-driven sediment loading of swales was observed over the first month post-fire and extensive snow drifts were formed in these swales each winter season during the study. In the first year, sediment yields from north- and south-facing aspects averaged 0.99–8.62 t ha−1 at the short-hillslope scale (~0.004 ha), 0.02–1.65 t ha−1 at the long-hillslope scale (0.02–0.46 ha), and 0.24–0.71 t ha−1 at the swale scale (0.65–3.42 ha), and watershed scale sediment yield was 2.47 t ha−1. By the second year post fire, foliar cover exceeded 120% across the site, but bare ground remained more than 60%. Sediment yield in the second year was greatly reduced across short- to long-hillslope scales (0.02–0.04 t ha−1), but was similar to first-year measures for swale plots (0.24–0.61 t ha−1) and at the watershed scale (3.05 t ha−1). Nearly all the sediment collected across all spatial scales was delivered during runoff events associated with cold-season hydrologic processes, including rain-on-snow, rain-on-frozen soils, and snowmelt runoff. Approximately 85–99% of annual sediment collected across all silt fence plots each year was from swales. The high levels of sediment delivered across hillslope to watershed scales in this study are attributed to observed preferential loading of fine sediments into swale channels by aeolian processes in the immediate post-fire period and subsequent flushing of these sediments by runoff from cold-season hydrologic processes. Our results suggest that the interaction of aeolian and cold-season hydrologic-driven erosion processes is an important component for consideration in post-fire erosion assessment and prediction and can have profound implications for soil loss from these ecosystems

Pre- and post-fire pollutant loads in an urban fringe watershed in Southern California

Post-fire runoff has the potential to be a large source of contaminants to downstream areas. However, the magnitude of this effect in urban fringe watersheds adjacent to large sources of airborne contaminants is not well documented. The current study investigates the impacts of wildfire on stormwater contaminant loading from the upper Arroyo Seco watershed, burned in 2009. This watershed is adjacent to the Greater Los Angeles, CA, USA area and has not burned in over 60 years. Consequently, it acts as a sink for regional urban pollutants and presents an opportunity to study the impacts of wildfire. Pre- and post-fire storm samples were collected and analyzed for basic cations, trace metals, and total suspended solids. The loss of vegetation and changes in soil properties from the fire greatly increased the magnitude of storm runoff, resulting in sediment-laden floods carrying high concentrations of particulate-bound constituents. Post-fire concentrations and loads were up to three orders of magnitude greater than pre-fire values for many trace metals, including lead and cadmium. A shift was also observed in the timing of chemical delivery, where maximum suspended sediment, trace metal, and cation concentrations coincided with, rather than preceded, peak discharge in the post-fire runoff, amplifying the fire's impacts on mass loading. The results emphasize the importance of sediment delivery as a primary mechanism for post-fire contaminant transport and suggest that traditional management practices that focus on treating only the early portion of storm runoff may be less effective following wildfire. We also advocate that watersheds impacted by regional urban pollutants have the potential to pose significant risk for downstream communities and ecosystems after fire. © 2013 Springer Science+Business Media Dordrecht