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Abstract: The SARS-CoV-2 pandemic has accelerated the development of virus concentration and molecular-based virus detection methods, monitoring systems and overall approach to epidemiology. Early into the pandemic, wastewater-based epidemiology started to be employed as a tool for tracking the virus transmission dynamics in a given area. The complexity of wastewater coupled with a lack of standardized methods led us to evaluate each step of the analysis individually and see which approach gave the most robust results for SARS-CoV-2 monitoring in wastewater. In this article, we present a step-by-step, retrospective view on the method development and implementation for the case of a pilot monitoring performed in Slovenia. We specifically address points regarding the thermal stability of the samples during storage, screening for the appropriate sample concentration and RNA extraction procedures and real-time PCR assay selection. Here, we show that the temperature and duration of the storage of the wastewater sample can have a varying impact on the detection depending on the structural form in which the SARS-CoV-2 target is present. We found that concentration and RNA extraction using Centricon filtration units coupled with Qiagen RNA extraction kit or direct RNA capture and extraction using semi-automated kit from Promega give the most optimal results out of the seven methods tested. Lastly, we confirm the use of N1 and N2 assays developed by the CDC (USA) as the best performing assays among four tested in combination with Fast Virus 1-mastermix. Data show a realistic overall process for method implementation as well as provide valuable information in regards to how different approaches in the analysis compare to one another under the specific conditions present in Slovenia during a pilot monitoring running from the beginning of the pandemic.
Keywords: waste water, method evolution, virus detection, SARS-CoV-2
Published in DiRROS: 17.07.2024; Views: 270; Downloads: 183
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Abstract: Tartary buckwheat flavonoids (TBFs) exhibit diverse biological activities, with antioxidant, antidiabetes, anti-inflammatory, and cholesterol-lowering properties. In this study, we investigated the role of TBFs in attenuating glucose and lipid disturbances in diabetic mice and hence preventing the occurrence of diabetes-related colon lesions in mice by regulating the gut microbiota. The results showed that TBFs (1) reversed blood glucose levels and body weight changes; (2) improved levels of serum total cholesterol (TC), triglycerides (TGs), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and fasting insulin; and (3) significantly reduced diabetes-related colon lesions in diabetic mice. In addition, TBFs also affected the diabetes-related imbalance of the gut microbiota and enriched beneficial microbiota, including Akkermansia and Prevotella. The TBF also selectively increased short-chain fatty acid-producing bacteria, including Roseburia and Odoribacter, and decreased the abundance of the diabetes-related gut microbiota, including Escherichia, Mucispirillum, and Bilophila. The correlation analysis indicated that TBFs improved metabolic parameters related to key communities of the gut microbiota. Our data suggested that TBFs alleviated glucose and lipid disturbances and improved colon lesions in diabetic mice, possibly by regulating the community composition of the gut microbiota. This regulation of the gut microbiota composition may explain the observed effects of TBFs to alleviate diabetes-related symptoms.
Keywords: Tartary buckwheat, flavonoids, microbiota, mice
Published in DiRROS: 17.07.2024; Views: 208; Downloads: 242
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Abstract: Coastal-estuarine habitats are rapidly changing due to global climate change, with impacts influenced by the variability of carbonate chemistry conditions. However, our understanding of the responses of ecologically and economically important calcifiers to pH variability and temporal variation is limited, particularly with respect to shell-building processes. We investigated the mechanisms driving biomineralogical and physiological responses in juveniles of introduced (Pacific; Crassostrea gigas) and native (Olympia; Ostrea lurida) oysters under flow-through experimental conditions over a six-week period that simulate current and future conditions: static control and low pH (8.0 and 7.7); low pH with fluctuating (24-h) amplitude (7.7 ± 0.2 and 7.7 ± 0.5); and high-frequency (12-h) fluctuating (8.0 ± 0.2) treatment. The oysters showed physiological tolerance in vital processes, including calcification, respiration, clearance, and survival. However, shell dissolution significantly increased with larger amplitudes of pH variability compared to static pH conditions, attributable to the longer cumulative exposure to lower pH conditions, with the dissolution threshold of pH 7.7 with 0.2 amplitude. Moreover, the high-frequency treatment triggered significantly greater dissolution, likely because of the oyster’s inability to respond to the unpredictable frequency of variations. The experimental findings were extrapolated to provide context for conditions existing in several Pacific coastal estuaries, with time series analyses demonstrating unique signatures of pH predictability and variability in these habitats, indicating potentially benefiting effects on fitness in these habitats. These implications are crucial for evaluating the suitability of coastal habitats for aquaculture, adaptation, and carbon dioxide removal strategies.
Keywords: ocean acidification, diel pH variability, amplitude, shell dissolution, predictability
Published in DiRROS: 17.07.2024; Views: 263; Downloads: 278
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Abstract: Marine primary producers are highly sensitive to environmental deterioration caused by natural and human-induced stressors. Following the Water Framework Directive and the Marine Strategy Framework Directive requirements, the importance of using the different primary producers of the coastal marine ecosystem (pelagic: phytoplankton and benthic: macroalgae and angiosperms) as appropriate tools for an integrated assessment of the ecological status of the coastal environment has been recognized. However, the processes by which water column characteristics and phytobenthic indicators are linked have not been systematically studied. Based on a large dataset from three Mediterranean sub-basins (Adriatic, Ionian and Aegean Seas) with different trophic conditions, this study aims to explore the coupled responses of benthic and pelagic primary producers to eutrophication pressures on a large scale, focusing on the structural and functional traits of benthic macroalgal and angiosperm communities, and to investigate the key drivers among the different eutrophication-related pelagic indicators (such as nutrient and Chl-a concentrations, water transparency, etc.) that can force the benthic system indicators to low ecological quality levels. In addition to the effects of high nutrient loading on phytoplankton biomass, our results also show that increased nutrient concentrations in seawater have a similar effect on macroalgal communities. Indeed, increasing nutrient concentrations lead to increased coverage of opportunistic macroalgal species at the expense of canopy-forming species. Most structural traits of Posidonia oceanica (expressed either as individual metrics: shoot density, lower limit depth and lower limit type, or in the context of PREI index) show opposite trends to increasing levels of pressure indicators such as ammonium, nitrate, phosphate, Chl-a and light attenuation. Furthermore, our results highlight the regulating effect of light availability on the ecological status of seagrass meadows (Posidonia oceanica and Cymodocea nodosa). Increasing leaf length values of C. nodosa are closely associated with higher turbidity values linked to higher phytoplankton biomass (expressed as Chl-a). Overall, the coupling of pelagic and benthic primary producers showed consistent patterns across trophic gradients at the subregional scale.
Keywords: macroalgae, seagrasses, phytoplankton, eutrophication, Mediterranean Sea
Published in DiRROS: 17.07.2024; Views: 300; Downloads: 181
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Abstract: We report here that synthetic cannabinoid WIN 55,212–2 inhibits tumor cell proliferation and induces cell death of oral and pancreatic tumor cells, and the effect is much more pronounced on stem-like/poorly differentiated OSCSCs and MP2 cells when compared to well-differentiated OSCCs, and PL-12 tumor cells. In addition, WIN 55,212-2 decreases cell surface expression of CD44, CD54, MHC class I and PD-L1 on oral and pancreatic tumor cells with the exception of PD-L1 expression on well-differentiated PL-12 pancreatic tumor cells which exhibits an increase in the expression rather than a decrease. Overall, we demonstrate that WIN 55,212-2 has an increased targeting activity against cancer stem cells/poorly differentiated oral and pancreatic tumor cells when compared to well-differentiated tumor cells, and furthermore, such differences in function do not correlate with the levels of CB1 and CB2 receptor expression on tumor cells, suggesting it's function either through post-receptor mediated activation and/or yet-to-be identified novel receptors. Intraperitoneal (IP) delivery of WIN 55-212-2 in humanized BLT mice is found to impart an activating potential for NK cells demonstrating increased NK cell mediated cytotoxicity and secretion of IFN-γ in our preliminary experiments. These results not only suggest a direct targeting of CSCs/poorly differentiated tumors by WIN 55-212-2 but also by indirect targeting of such tumors through the activation and increased functions of NK cells.
Keywords: cancer stem cells, cannabinoids, cell death, cancer biology, genetic toxicology
Published in DiRROS: 17.07.2024; Views: 301; Downloads: 180
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