Road dust resuspension is one of the main sources of particulate matter with impacts on air quality, health and climate. With the aim of characterising the thoracic fraction, a portable resuspension ...chamber was used to collect road dust from five main roads in Oporto and an urban tunnel in Braga, north of Portugal. The PM10 samples were analysed for: i) carbonates by acidification and quantification of the evolved CO2, ii) carbonaceous content (OC and EC) by a thermo-optical technique, iii) elemental composition by ICP-MS and ICP-AES after acid digestion, and iv) organic speciation by GC–MS. Dust loadings of 0.48±0.39mgPM10m−2 were obtained for asphalt paved roads. A much higher mean value was achieved in a cobbled pavement (50mgPM10m−2). In general, carbonates were not detected in PM10. OC and EC accounted for PM10 mass fractions up to 11% and 5%, respectively. Metal oxides accounted for 29±7.5% of the PM10 mass from the asphalt paved roads and 73% in samples from the cobbled street. Crustal and anthropogenic elements, associated with tyre and brake wear, dominated the inorganic fraction. PM10 comprised hundreds of organic constituents, including hopanoids, n-alkanes and other aliphatics, polycyclic aromatic hydrocarbons (PAH), alcohols, sterols, various types of acids, glycerol derivatives, lactones, sugars and derivatives, phenolic compounds and plasticizers. In samples from the cobbled street, these organic classes represented only 439μgg−1PM10, while for other pavements mass fractions up to 65mgg−1PM10 were obtained. Except for the cobbled street, on average, about 40% of the analysed organic fraction was composed of plasticizers. Although the risk via inhalation of PAH was found to be insignificant, the PM10 from some roads can contribute to an estimated excess of 332 to 2183 per million new cancer cases in adults exposed via ingestion and dermal contact.
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•Dust loadings<1mgPM10m−2 were obtained for asphalt paved roads.•These values are lower than those reported for other cities.•Dust loadings about 50 times higher were found for a cobbled pavement.•OC & EC accounted for small PM10 mass fractions, while inorganic components dominate.•OC comprised hundreds of organic constituents; plasticizers were the most abundant.
Calcium carbonates (CaCO
) often accumulate in mangrove and seagrass sediments. As CaCO
production emits CO
, there is concern that this may partially offset the role of Blue Carbon ecosystems as CO
...sinks through the burial of organic carbon (C
). A global collection of data on inorganic carbon burial rates (C
, 12% of CaCO
mass) revealed global rates of 0.8 TgC
yr
and 15-62 TgC
yr
in mangrove and seagrass ecosystems, respectively. In seagrass, CaCO
burial may correspond to an offset of 30% of the net CO
sequestration. However, a mass balance assessment highlights that the C
burial is mainly supported by inputs from adjacent ecosystems rather than by local calcification, and that Blue Carbon ecosystems are sites of net CaCO
dissolution. Hence, CaCO
burial in Blue Carbon ecosystems contribute to seabed elevation and therefore buffers sea-level rise, without undermining their role as CO
sinks.
Until there is an effective implementation of COVID-19 vaccination program, a robust testing strategy, along with prevention measures, will continue to be the most viable way to control disease ...spread. Such a strategy should rely on disparate diagnostic tests to prevent a slowdown in testing due to lack of materials and reagents imposed by supply chain problems, which happened at the beginning of the pandemic. In this study, we have established a single-tube test based on RT-LAMP that enables the visual detection of less than 100 viral genome copies of SARS-CoV-2 within 30 min. We benchmarked the assay against the gold standard test for COVID-19 diagnosis, RT-PCR, using 177 nasopharyngeal RNA samples. For viral loads above 100 copies, the RT-LAMP assay had a sensitivity of 100% and a specificity of 96.1%. Additionally, we set up a RNA extraction-free RT-LAMP test capable of detecting SARS-CoV-2 directly from saliva samples, albeit with lower sensitivity. The saliva was self-collected and the collection tube remained closed until inactivation, thereby ensuring the protection of the testing personnel. As expected, RNA extraction from saliva samples increased the sensitivity of the test. To lower the costs associated with RNA extraction, we performed this step using an alternative protocol that uses plasmid DNA extraction columns. We also produced the enzymes needed for the assay and established an in-house-made RT-LAMP test independent of specific distribution channels. Finally, we developed a new colorimetric method that allowed the detection of LAMP products by the visualization of an evident color shift, regardless of the reaction pH.
The impact of inhalable fine particulate matter (PM2.5, aerodynamic diameter <2.5 μm) on public health is of great concern worldwide. Knowledge on their harmful effects are mainly due to studies ...carried out with whole air particles, with the contribution of their different fractions remaining largely unknown. Herein, a set of urban PM2.5 samples were collected during daytime and nighttime periods in autumn and spring, aiming to address the seasonal and day-night variability of water-soluble organic matter (WSOM) composition. In vitro analysis of the oxidative and pro-inflammatory potential of WSOM samples was carried out in both acute (24 h) and chronic (3 weeks) exposure setups using Raw264.7 macrophages as cell model. Findings revealed that the structural composition of WSOM samples differs between seasons and in a day–night cycle. Cell exposure resulted in an increase in the transcription of the cytoprotective Hmox1 and pro-inflammatory genes Il1b and Nos2, leading to a moderate pro-inflammatory status. These macrophages showed an impaired capacity to subsequently respond to a strong pro-inflammatory stimulus such as bacterial lipopolysaccharide, which may implicate a compromised capacity to manage harmful pathogens. Further investigation on aerosol WSOM could help to constrain the mechanisms of WSOM-induced respiratory diseases and contribute to PM2.5 regulations.
MicroRNAs (miRNAs) are aberrantly expressed in human cancer and involved in the (dys)regulation of cell survival, proliferation, differentiation and death. Specifically, miRNA-143 (miR-143) is ...down-regulated in human colon cancer. In the present study, we evaluated the role of miR-143 overexpression on the growth of human colon carcinoma cells xenografted in nude mice (immunodeficient mouse strain: N: NIH(s) II-nu/nu).
HCT116 cells with stable miR-143 overexpression (Over-143) and control (Empty) cells were subcutaneously injected into the flanks of nude mice, and tumor growth was evaluated over time. Tumors arose ∼ 14 days after tumor cell implantation, and the experiment was ended at 40 days after implantation. miR-143 was confirmed to be significantly overexpressed in Over-143 versus Empty xenografts, by TaqMan® Real-time PCR (p<0.05). Importantly, Over-143 xenografts displayed slower tumor growth compared to Empty xenografts from 23 until 40 days in vivo (p<0.05), with final volumes of 928±338 and 2512±387 mm(3), respectively. Evaluation of apoptotic proteins showed that Over-143 versus Empty xenografts displayed reduced Bcl-2 levels, and increased caspase-3 activation and PARP cleavage (p<0.05). In addition, the incidence of apoptotic tumor cells, assessed by TUNEL, was increased in Over-143 versus Empty xenografts (p<0.01). Finally, Over-143 versus Empty xenografts displayed significantly reduced NF-κB activation and ERK5 levels and activation (p<0.05), as well as reduced proliferative index, evaluated by Ki-67 immunohistochemistry (p<0.01).
Our results suggest that reduced tumor volume in Over-143 versus Empty xenografts may result from increased apoptosis and decreased proliferation induced by miR-143. This reinforces the relevance of miR-143 in colon cancer, indicating an important role in the control of in vivo tumor progression, and suggesting that miR-143 may constitute a putative novel therapeutic tool for colon cancer treatment that warrants further investigation.
Aims
Brazil ranks high in the number of coronavirus disease 19 (COVID‐19) cases and the COVID‐19 mortality rate. In this context, autopsies are important to confirm the disease, determine associated ...conditions, and study the pathophysiology of this novel disease. The aim of this study was to assess the systemic involvement of COVID‐19. In order to follow biosafety recommendations, we used ultrasound‐guided minimally invasive autopsy (MIA‐US), and we present the results of 10 initial autopsies.
Methods and results
We used MIA‐US for tissue sampling of the lungs, liver, heart, kidneys, spleen, brain, skin, skeletal muscle and testis for histology, and reverse transcription polymerase chain reaction to detect severe acute respiratory syndrome coronavirus 2 RNA. All patients showed exudative/proliferative diffuse alveolar damage. There were intense pleomorphic cytopathic effects on the respiratory epithelium, including airway and alveolar cells. Fibrinous thrombi in alveolar arterioles were present in eight patients, and all patients showed a high density of alveolar megakaryocytes. Small thrombi were less frequently observed in the glomeruli, spleen, heart, dermis, testis, and liver sinusoids. The main systemic findings were associated with comorbidities, age, and sepsis, in addition to possible tissue damage due to the viral infection, such as myositis, dermatitis, myocarditis, and orchitis.
Conclusions
MIA‐US is safe and effective for the study of severe COVID‐19. Our findings show that COVID‐19 is a systemic disease causing major events in the lungs and with involvement of various organs and tissues. Pulmonary changes result from severe epithelial injury and microthrombotic vascular phenomena. These findings indicate that both epithelial and vascular injury should be addressed in therapeutic approaches.
Background & Aims Non-alcoholic fatty liver disease (NAFLD) comprises a spectrum of stages from simple steatosis to non-alcoholic steatohepatitis (NASH). However, disease pathogenesis remains largely ...unknown. microRNA (miRNA or miR) expression has recently been reported to be altered in human NASH, and modulated by ursodeoxycholic acid (UDCA) in the rat liver. Here, we aimed at evaluating the miR-34a/Sirtuin 1(SIRT1)/p53 pro-apoptotic pathway in human NAFLD, and to elucidate its function and modulation by UDCA in the rat liver and primary rat hepatocytes. Methods Liver biopsies were obtained from NAFLD morbid obese patients undergoing bariatric surgery. Rat livers were collected from animals fed a 0.4% UDCA diets. Primary rat hepatocytes were incubated with bile acids or free fatty acids (FFAs) and transfected with a specific miRNA-34a precursor and/or with a p53 overexpression plasmid. p53 transcriptional activity was assessed by ELISA and target reporter constructs. Results miR-34a, apoptosis and acetylated p53 increased with disease severity, while SIRT1 diminished in the NAFLD liver. UDCA inhibited the miR-34a/SIRT1/p53 pathway in the rat liver in vivo and in primary rat hepatocytes. miR-34a overexpression confirmed its targeting by UDCA, which prevented miR-34a-dependent repression of SIRT1, p53 acetylation, and apoptosis. Augmented apoptosis by FFAs in miR-34a overexpressing cells was also inhibited by UDCA. Finally, p53 overexpression activated miR-34a/SIRT1/p53, which in turn was inhibited by UDCA, via decreased p53 transcriptional activity. Conclusions Our results support a link between liver cell apoptosis and miR-34a/SIRT1/p53 signaling, specifically modulated by UDCA, and NAFLD severity. Potential endogenous modulators of NAFLD pathogenesis may ultimately provide new tools for therapeutic intervention.
In both Run 1 and Run 2 of the LHC, the CMS collaboration has observed an excess of events in the searches for low-mass Higgs bosons in the diphoton final state at a mass of about 95 GeV. After a ...recent update of the experimental analysis, in which the full Run 2 data collected at 13 TeV has been included and an improved experimental calibration has been applied, the local significance of the excess amounts to 2
.
9
σ
. The presence of this diphoton excess is especially interesting in view of a further excess observed by CMS in ditau final states at a comparable mass and similar local significance. Moreover, an excess of events with about 2
σ
local significance and consistent with a mass of 95 GeV was observed in LEP searches for a Higgs boson decaying to pairs of bottom quarks. We interpret the CMS diphoton excess in combination with the ditau excess in terms of a pseudoscalar resonance in the CP-conserving two-Higgs-doublet model (2HDM). Furthermore, we discuss the possibility that, if CP-violation is taken into account, a CP-mixed scalar state can in addition describe the LEP result, thus accommodating all three excesses simultaneously. We find that the region of parameter space where both the CMS diphoton and ditau excesses can be fitted is in tension with current constraints from the flavour sector, potentially calling for other new-physics contributions to flavour-physics observables, most notably
b
→
sγ
transitions. Additionally, the new source of CP-violation violates electron EDM constraints by about an order of magnitude. We also comment on the compatibility with the recent ATLAS diphoton searches.
: Galacto‐oligosaccharides (GOS) have now been definitely established as prebiotic ingredients after in vitro and animal and human in vivo studies. Currently, GOS are produced by glycoside ...hydrolases (GH) using lactose as substrate. Converting lactose into GOS by GH results in mixtures containing GOS of different degrees of polymerization (DP), unreacted lactose, and monomeric sugars (glucose and galactose). Recent and future developments in the production of GOS aim at delivering purer and more efficient mixtures. To produce high‐GOS‐content mixtures, GH should not only have good ability to catalyze the transgalactosylation reaction relative to hydrolysis, but also have low affinity for the GOS formed relative to the affinity for lactose. In this article, several microbial GH, proposed for the synthesis of GOS, are hierarchized according to the referred performance indicators. In addition, strategies for process improvement are discussed. Besides the differences in purity of GOS mixtures, differences in the position of the glycosidic linkages occur, because different enzymes have different regiochemical selectivity. Depending on oligosaccharide composition, GOS products will vary in terms of prebiotic activity, as well as other physiological effects. This review focuses on GOS production from synthesis to purification processes. Physicochemical characteristics, physiological effects, and applications of these prebiotic ingredients are summarized. Regulatory aspects of GOS‐containing food products are also highlighted with emphasis on the current process of health claims evaluation in Europe.