Food is not only the source of nutrition for humans but also plays various roles in our daily lives, beliefs, and relationship. In China, one of the fundamental cultural elements is the sharing of food. Typically, the courses are served in the center of the table from which guests serve themselves on their plates or serve guests using their chopsticks. With the COVID-19 outbreak, people were advised to separate dining or at least use gongshao 公勺'serving spoon' or gongkuai公筷'serving chopsticks' instead of picking food directly from serving plates with their own chopsticks. The “table revolution” is a crucial issue: if it succeeds, it will change China's face. Public advertisements, as giant billboards on Shanghai's streets talking of serving chopsticks as a way to set the heart at ease, showed slogans like: "The distance between you and civilized dining is just one pair of serving chopsticks". Nevertheless, serving chopsticks have not quite caught on yet in China as they have done in Taiwan and Japan. According to the survey from Ma Lihua et al. (2020) resistance is strong. In a declaration from China Hotel Association, we find out: "Some restaurants in China have provided individual meals and public chopsticks and spoons for decades, but not everyone chooses to use them due to traditional eating habits". According to the Global Times, "if they eat with close friends and relatives, they would feel too embarrassed to use serving chopsticks as it seems like they dislike sharing with others, which often makes people uncomfortable" (Li Lei, Zhang Hu, Global Times 2020). The New York Times adds: "Many see sharing food with one's own chopsticks as among the most authentic expressions of China's communal culture and emphasis on family, no less integral than hugging is to Americans or the cheek kiss is to the French". The “dining table revolution”, through the lens of newspapers, is going to be an uphill battle.

As the novel coronavirus disease sweeps across the world, there is growing speculation on the role that atmospheric factors may have played on the different distribution of SARS-CoV-2, and on the epidemiological characteristics of COVID-19. Knowing the role that environmental factors play in influenza virus outbreaks, environmental pollution and, in particular, atmospheric airborne (particulate matter, PM) has been considered as a potential key factor in the spread and mortality of COVID-19. A possible role of the PM as the virus carrier has also been debated. The role of PM in exacerbating respiratory and cardiovascular disease has been well recognized. Accumulating evidence support the hypothesis that PM can trigger inflammatory response at molecular, cellular and organ levels. On this basis, we developed the hypothesis that PM may play a role as a booster of COVID-19 rather than as a carrier of SARS-CoV-2. To support our hypothesis, we analyzed the molecular signatures detected in cells exposed to PM samples collected in one of the most affected areas by the COVID-19 outbreak, in Italy. T47D human breast adenocarcinoma cells were chosen to explore the global gene expression changes induced by the treatment with organic extracts of PM 2.5. The analysis of the KEGG's pathways showed modulation of several gene networks related to the leucocyte transendothelial migration, cytoskeleton and adhesion system. Three major biological process were identified, including coagulation, growth control and immune response. The analysis of the modulated genes gave evidence for the involvement of PM in the endothelial disease, coagulation disorders, diabetes and reproductive toxicity, supporting the hypothesis that PM, directly or through molecular interplay, affects the same molecular targets as so far known for SARS-COV-2, contributing to the cytokines storm and to the aggravation of the symptoms triggered by COVID-19. We provide evidence for a plausible cooperation of receptors and transmembrane proteins, targeted by PM and involved in COVID-19, together with new insights into the molecular interplay of chemicals and pathogens that could be of importance for sustaining public health policies and developing new therapeutic approaches.

We present intermediate-resolution (R$\sim$1000) spectra in the $\sim$3500-10,000 A range of 14 globular clusters in the magellanic irregular galaxy NGC 4449 acquired with the Multi Object Double Spectrograph on the Large Binocular Telescope. We derived Lick indices in the optical and the CaII-triplet index in the near-infrared in order to infer the clusters' stellar population properties. The inferred cluster ages are typically older than $\sim$9 Gyr, although ages are derived with large uncertainties. The clusters exhibit intermediate metallicities, in the range $-1.2\lesssim$[Fe/H]$\lesssim-0.7$, and typically sub-solar [$\alpha/Fe$] ratios, with a peak at $\sim-0.4$. These properties suggest that i) during the first few Gyrs NGC 4449 formed stars slowly and inefficiently, with galactic winds having possibly contributed to the expulsion of the $\alpha$-elements, and ii) globular clusters in NGC 4449 formed relatively "late", from a medium already enriched in the products of type Ia supernovae. The majority of clusters appear also under-abundant in CN compared to Milky Way halo globular clusters, perhaps because of the lack of a conspicuous N-enriched, second-generation of stars like that observed in Galactic globular clusters. Using the cluster velocities, we infer the dynamical mass of NGC 4449 inside 2.88 kpc to be M($<$2.88 kpc)=$3.15^{+3.16}_{-0.75} \times 10^9~M_\odot$. We also report the serendipitous discovery of a planetary nebula within one of the targeted clusters, a rather rare event.

The evaluation of chemical hazard is based on the identification of the quality and the quantity of adverse effects as a consequence of exposure. The adverse effects that do not involve genetic damage are often related to the chemical dose or concentration. The adverse outcome is the consequence of a row of key events, each targeting a different biological trait. The identification of these key events at molecular and cellular level would provide novel biomarkers of exposure and risk. The application of toxicogenomics approaches to experimental models of chemical exposure allows the detection of gene pathways involved in response to low doses of the chemical as an early endpoint of adversity. The use of toxicogenomics would improve the knowledge on the dose-response relationship, linking the environmental exposure to the effect on the population and allowing a better refinement of the quantitative risk assessment. In this context, the gene modulation data can be used to calculate a No Observed Transcriptional Effect Level (NOTEL). In this paper we present a method for evaluating the NOTEL based on anomaly detection: a classifier is built that discriminates between target class instances, i.e., normal cases, and anomalies, i.e., samples with significant transcriptional effects. The strength of this method is that (i) it can be applied to cases in which few samples are available and the space dimension is high and (ii) it makes use of the complete gene expression profiles. (C) 2017 Elsevier B.V. All rights reserved.

The particulate matter represents one of the most complex environmental mixtures, whose effects on human health and environment vary according to particles characteristics and source of emissions. The present study describes an integrated approach, including in vitro tests and toxicogenomics, to highlight the effects of air particulate matter on toxicological relevant endpoints.Air samples (PM2.5) were collected in summer and winter at different sites, representative of different levels of air pollution. Samples organic extracts were tested in the BALB/c 3T3 CTA at a dose range 1-12 m(3). The effect of the exposure to the samples at a dose of 8 m(3) on the whole-genome transcriptomic profile was also assessed.All the collected samples induced dose-related toxic effects in the exposed cells. The modulated gene pathways confirmed that toxicity was related to sampling season and sampling site. The analysis of the KEGG's pathways showed modulation of several gene networks related to oxidative stress and inflammation. Even if the samples did not induce cell transformation in the treated cells, gene pathways related to the onset of cancer were modulated as a consequence of the exposure.This integrated approach could provide valuable information for predicting toxic risks in humans exposed to air pollution. (C) 2014 Elsevier Ltd. All rights reserved.

Penconazole is a systemic triazole fungicide mainly used on grapes. The UE Maximum Residue Level (MRL) for penconazole is set at 0.2 ppm in wine and grapes. In the aim of identifying potential biomarkers of exposure to penconazole and possibly highlighting its endocrine disrupting mode of action, we used a transcriptomics-based approach to detect genes, that are transcriptionally modulated by penconazole, by using an appropriate in vitro model. T-47D cells were treated with commercial penconazole or penconazole contaminated grape extracts for 4 h at doses close to the MRL. The whole-genome transcriptomic profile was assessed by using genome 44 K oligo-microarray slides. The list of common genes generated by the two treatments could be representative of potential markers of exposure. In order to understand the role of these genes in key events related to adversity, a pathway analysis was performed on a list of genes with the same modulation trend (up or down). The analysis returned a set of genes involved in Thyroid Cancer Pathway, thus confirming a role of penconazole in endocrine disrupting mediated effects and strongly suggesting a possible mode of action in thyroid carcinogenesis.

Alternative methods to animal testing are considered as promising tools to support the prediction of toxicological risks from environmental exposure. Among the alternative testing methods, the cell transformation assay (CTA) appears to be one of the most appropriate approaches to predict the carcinogenic properties of single chemicals, complex mixtures and environmental pollutants. The BALB/c 3T3 CTA shows a good degree of concordance with the in vivo rodent carcinogenesis tests. Whole-genome transcriptomic profiling is performed to identify genes that are transcriptionally regulated by different kinds of exposures. Its use in cell models representative of target organs may help in understanding the mode of action and predicting the risk for human health. Aiming at associating the environmental exposure to health-adverse outcomes, we used an integrated approach including the 3T3 CTA and transcriptomics on target cells, in order to evaluate the effects of airborne particulate matter (PM) on toxicological complex endpoints. Organic extracts obtained from PM2.5 and PM1 samples were evaluated in the 3T3 CTA in order to identify effects possibly associated with different aerodynamic diameters or airborne chemical components. The effects of the PM2.5 extracts on human health were assessed by using whole-genome 44 K oligo-microarray slides. Statistical analysis by GeneSpring GX identified genes whose expression was modulated in response to the cell treatment. Then, modulated genes were associated with pathways, biological processes and diseases through an extensive biological analysis. Data derived from in vitro methods and omics techniques could be valuable for monitoring the exposure to toxicants, understanding the modes of action via exposure-associated gene expression patterns and to highlight the role of genes in key events related to adversity.

The novel brominated flame retardants (NBFRs), 2-ethylhexyl-2,3,4,5-tetrabromobenzoate (TBB), Bis(2-ethylhexyl)-2,3,4,5-tetrabromophtalate (TBPH), and 1,2,5,6-tetrabromocyclooctane (TBCO) are components of flame retardant mixtures including Firemaster 550 and Saytex BC-48. Despite the detection of these NBFRs in environmental and biotic matrices, studies regarding their toxicological effects are poorly represented in the literature. The present study examined endocrine disruption by these three NBFRs using the yeast YES/YAS reporter assay and the mammalian H295R steroidogenesis assay. Activation of the aryl hydrocarbon receptor (AhR) was also assessed using the H4IIE reporter assay. The NBFRs produced no TCDD-like effects in the H4IIE assay or agonistic effects in the YES/YAS assays. TBB produced a maximal antiestrogenic effect of 62% at 0.5 mg L−1 in the YES assay while TBPH and TBCO produced maximal antiandrogenic effects of 74% and 59% at 300 mg L−1 and 1500 mg L−1, respectively, in the YAS assay. Significant effects were also observed in the H295R assay. At 0.05 mg L−1, 15 mg L−1, and 15 mg L−1 TBB, TBPH, and TBCO exposures, respectively resulted in a 2.8-fold, 5.4-fold, and 3.3-fold increase in concentrations of E2. This is one of the first studies to demonstrate the in vitro endocrine disrupting potentials of TBB, TBPH, and TBCO.

Cell transformation assays (CTAs) are currently regarded as the only possible in vitro alternative to animal testing for carcinogenesis studies. CTAs have been proposed as screening tests for the carcinogenic potential of compounds that have no evidence of genotoxicity but present structural alerts for carcinogenicity. We have extensively used the BALB/c 3T3 model based on the A31 cell clone to test single chemicals, complex mixtures and environmental pollutants. In the prevalidation study carried out by ECVAM, the improved protocol is based on BALB/c 3T3 A31-1-1 cells, a clone derived by A31 cells, that is very sensitive to PAH-induced transformation. The present study was performed in the aim to compare the results obtained with the two different clones exposed to different classes of carcinogens. Cells were treated with PAHs (3-methylcholanthrene, benzo(a)pyrene), alkylating agents (melphalan) and aloethanes (1,2-dibromoethane). The induction of cytotoxicity and the onset of chemically transformed foci were evaluated by two experimental protocols, differing for cell seeding density and chemical treatment duration. The A31-1-1 cells showed higher inherent transformation rate after PAHs treatment, but they were insensitive to 1,2-dibromoethane at concentrations that usually induced transformation in A31 cells. As 1,2-dibromoethane is bioactivated to reactive forms able to bind DNA mainly through the conjugation with intracellular glutathione, these results suggested a reduced activity of phase-2 enzymes involved in glutathione conjugation in A31-1-1 cells. Our results give evidence that inherent metabolic capacity of cells may play a critical role in in vitro cell transformation, cautioning against possible misclassification of chemicals.

The risk of human exposure to environmental compounds has historically been based on the extrapolation of data from animal studies performed at high levels of exposure to single chemicals. Models used for this extrapolation include the assumption of low-dose linearity for genotoxic carcinogens and a predictable threshold for non mutagenic compounds.This concept is increasingly questioned as the theory of hormesis is more and more supported by experimental evidence. Hormesis is a term to define the biphasic dose-response to an environmental agent that can induce stimulatory or beneficial effects at low doses and inhibitory or adverse effects at high doses. The adaptative response of living organisms to the environment poses serious questions about the reliability of the current approach for risk assessment. Some other considerations should be taken in account when defining the effects of chemicals on human health. The human population is chronically exposed to low doses of environmental pollutants. Environmental and genetic factors both play a role in the development of most diseases. Environment include any external agent, which also means food and medications. The interactions among all these aspects make the comprehension of the whole picture difficult.The sequencing of the human genome has provided new opportunities, new technologies and new tools to study the environmental impact on living organisms and life processes. Toxicogenomics has the potential to provide all the information we need for the risk prediction in just one experimental design. Toxicogenomic technologies can be used for screening the toxic potential of environmental agents, for assessing cellular responses to different doses, for classifying chemicals on the basis of their mechanism of action, for monitoring human exposure and for predicting individual variability in the response to toxicants. We used a combination of in vitro models and in vivo studies to identify expression signatures of low-dose exposures to physical and chemical agents, highlighting hormetic effects in different target and endpoints. omics approaches, functional analysis studies, which can be used alone or in combination, could provide information for different exposure in different scenarios.By using such an approach we performed in vitro and in vivo studies and were able to profile the response to the exposure to low doses of ionizing radiation at cellular and molecular level, by defining a set of genes whose expression is directly related to the exposure (paper in preparation).

The prediction of the carcinogenic risk for humans is mostly based on animal experiments. For the last 20 years, however, the scientific community has paid great attention to alternative strategies in compliance with common moral and ethical values. The new European chemical regulation REACH (Reg. EC 1907/2006) requires the performance of new studies in vertebrates only as a last resort. REACH asks for the development of validated in vitro protocols that can replace, in the medium to the long term, animal bioassays. An in vitro cell transformation assay (CTA) is proposed as an alternative to in vivo carcinogenicity testing. This assay is reported in the list of accepted methods for REACH (Reg. EC 440/2008). The BALB/c 3T3 model represents one of the most well-known CTAs and is regarded as a useful tool to screen single chemicals or complex mixtures for carcinogenicity prediction. In this study we used a modified protocol to highlight the transforming potential of three single compounds, ethinylestradiol (EE), azathioprine (AZA-T), melphalan, and two polychlorinated biphenyls (PCBs) mixtures, which are known or suspected to be human carcinogens. We also evaluated the activity of the antioxidant alpha-lipoic acid (ALA), a promising tumor chemopreventive. A significant increase in transformation frequency was observed when the BALB/c 3T3 cells were exposed to EE, AZA-T or melphalan as well as after PCBs treatment. On the contrary, ALA did not induce any increase of foci occurrence. Our results confirm the suitability of the improved protocol to discriminate carcinogenic compounds and support the use of BALB/c 3T3 cell transformation assay as a possible alternative to predict carcinogenic risk to humans.

Recent whole-genome studies and in-depth expressed sequence tag (EST) analyses have identified most of the developmentally relevant genes in the urochordate, Ciona intestinalis. In this study, we made use of a large-scale oligo-DNA microarray to further investigate and identify genes with specific or correlated expression profiles, and we report global gene expression profiles for about 66% of all the C. intestinalis genes that are expressed during its life cycle. We succeeded in categorizing the data set into 5 large clusters and 49 sub-clusters based on the expression profile of each gene. This revealed the higher order of gene expression profiles during the developmental and aging stages. Furthermore, a combined analysis of microarray data with the EST database revealed the gene groups that were expressed at a specific stage or in a specific organ of the adult. This study provides insights into the complex structure of ascidian gene expression, identifies co-expressed gene groups and marker genes and makes predictions for the biological roles of many uncharacterized genes. This large-scale oligo-DNA microarray for C. intestinalis should facilitate the understanding of global gene expression and gene networks during the development and aging of a basal chordate.

The use of nuclear resources for medical purposes causes considerable concern about occupational exposure. Nevertheless, little information is available regarding the effects of low-dose irradiations protracted over time. We used oligomicroarrays to identify the genes that are transcriptionally regulated by persistent exposure to extremely low doses of ionizing radiation in 28 exposed professionals (mean cumulative effective dose +/- SD, 19 +/- 38 mSv) compared with a matched sample of nonexposed subjects. We identified 256 modulated genes from peripheral blood mononuclear cells profiles, and the main biological processes we found were DNA packaging and mitochondrial electron transport NADH to ubiquinone. Next we investigated whether a different pattern existed when only 22 exposed subjects with accumulated doses >2.5 mSv, a threshold corresponding to the natural background radiation in Italy per year, and mean equal to 25 +/- 41 mSv were used. In addition to DNA packaging and NADH dehydrogenase function, the analysis of the higher-exposed subgroup revealed a significant modulation of ion homeostasis and programmed cell death as well. The changes in gene expression that we found suggest different mechanisms from those involved in high-dose studies that may help to define new biomarkers of radiation exposure for accumulated doses below 25 mSv.

Background The clinical efficacy of camptothecin (CPT), a drug specifically targeting topoisomerase I (TopoI), is under evaluation for the treatment of malignant gliomas. Due to the high unresponsiveness of these tumours to chemotherapy, it would be very important to study the signalling network that drives camptothecin outcome in this type of cancer cells. To address this issue, we had previously compared the expression profile of human U87-MG glioblastoma cells with that of a CPT-resistant counterpart, giving evidence that the development of a robust inflammatory response was the main transcriptional effect associated with CPT resistance. Here we report time-related changes and cell line specific patterns of gene expression after CPT treatment by using two p53 wild-type glioblastoma cell lines, U87-MG and DBTRG-05, with different sensitivities to TopoI inhibition. Results First, we demonstrated that CPT treatment brings the two cell lines to completely different outcomes: accelerated senescence in U87-MG and apoptosis in DBTRG-05 cells. Then, to understand the different susceptibility to CPT, we used oligo-microarray to identify the genes whose expression was regulated during a time-course treatment, ranging from 2 h to 72 h. The statistical analysis of microarray data by MAANOVA (MicroArray ANalysis Of VAriance) showed much less modulated genes in apoptotic DBTRG-05 cells (155) with respect to the senescent U87-MG cells (3168), where the number of down-regulated genes largely exceeded that of the up-regulated ones (80% vs. 20%). Despite this great difference, the two data-sets showed a large overlapping (60% circa) mainly due to the expression of early stress responsive genes. The use of High-Throughput GoMINER and EASE tools, for functional analysis of significantly enriched GO terms, highlighted common cellular processes and showed that U87-MG and DBTRG-05 cells shared many GO terms, which are related to the down-regulation of cell cycle and mitosis and to the up-regulation of cell growth inhibition and DNA damage. Furthermore, the down-regulation of MYC and DP1 genes, which act as key transcription factors in cell growth control, together with the inhibition of BUB1, BUB3 and MAD2 mRNAs, which are known to be involved in the spindle checkpoint pathway, were specifically associated with the execution of senescence in U87-MG cells and addressed as critical factors that could drive the choice between different CPT-inducible effectors programs. In U87-MG cells we also found inflammation response and IL1-beta induction, as late transcriptional effects of Topo I treatment but these changes were only partially involved in the senescence development, as shown by IL1-beta gene silencing. Conclusion By comparing the transcription profile of two glioblastoma cell lines treated with camptothecin, we were able to identify the common cellular pathways activated upon Topo I inhibition. Moreover, our results helped in identifying some key genes whose expression seemed to be associated with the execution of senescence or apoptosis in U87-MG and DBTRG-05 cells, respectively.

3122 The manipulation of nuclear resources for medical purposes has a considerable concern in occupational exposure but the health risk due to low-dose ionizing radiation is still debated. Functional genomics approaches provide an opportunity to explore further complex biological processes and may lead to an increased understanding of the pathways that are induced by ionizing irradiation (IR). We used 60-mer oligo microarrays to identify genes that are transcriptionally regulated by low-dose IR in occupationally exposed workers. To this purpose, 28 exposed professional were compared with a reference sample of non-exposed people, working at the same place at a large health care facility. The expression profile of their peripheral blood lymphocytes (PBMC) was investigated according to a balanced block design, consisting in the equal distribution of cyanine 5 and cyanine 3 labels within the two classes of exposed and control samples. The raw intensity data derived from Feature Extraction software 7.5 were analysed by MAANOVA (MicroArray ANalysis Of VAriance) data analysis package of R programming environments and we carried out hypothesis testing using F statistics computed on the James-Stein shrinkage estimates of the error variance. By this approach, we identified 270 genes which were correlated to occupational low-dose IR exposure, including chromatin assembly genes as well as genes involved in apoptosis, DNA repair and response to stress. Since exposed individuals were recruited from two distinct departments and showed different ranges of accumulated doses, we furthermore performed a MAANOVA analysis to determine the differentially expressed genes discriminating between these two divisions. In this case, a list of 284 genes was obtained and the use of agglomerative hierarchical clustering with Ward’s linkage partly confirmed the homogeneity of the two sets of exposed individuals. Some of the genes, that were found to be differentially expressed between the workers of the two departments, were under the direct control of p53, such as p21-Waf1, ATF3 or TNFRSF10B. Moreover, beside chromatin assembly,which was strongly affected, many regulated genes were specifically involved in mRNA processing and protein biosynthesis, as well. In conclusion, we assessed the expression profile of peripheral blood lymphocytes from radio-exposed workers and identified a 270-gene set, referred to the whole sample of 28 exposed individuals, and an additional list of 284 genes that potentially serve as a signature for workers showing different cumulative doses. We aimed at providing further insight into the transcriptional fingerprint of low-dose IR in humans in order to improve the classical methods to estimate IR exposure and add information to the current models of the risk assessment for chronic low-dose exposure. This work was partly supported by the Italian Ministry of Health, Research Grants 2003