Association between heavy metal exposure and biomarkers for non-alcoholic fatty liver disease in Korean adolescents.

Objectives: The global prevalence of non-alcoholic fatty liver disease (NAFLD) in adolescents has increased. In addition to childhood obesity, environmental risk factors, such as heavy metals that are known to be involved in hepatotoxicity, play role in NAFLD occurrence. However, their association with NAFLD remains unclear. This study aimed to investigate the association between heavy metal exposure and NAFLD biomarkers in adolescents. Methods: In this cross-sectional study, we used the data of a total of 1505 adolescents aged 12-17 years who participated in the Korean National Environmental Health Survey III (2015-2017) and IV (2018-2020). The presence of blood lead (BPb), blood mercury (BHg), urinary mercury (UHg), and urinary cadmium (UCd) were measured. Liver enzymes including serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), and gamma-glutamyl transferase (GGT) were evaluated. For NAFLD biomarkers, the hepatic steatosis index (HSI) was calculated. Multivariate linear regression models, weighted quantile sum (WQS) regression, and Bayesian kernel machine regression (BKMR) model were used to investigate the association between heavy metals and NAFLD biomarkers. Results: Among heavy metals, mercury presence showed a significant association with NAFLD biomarkers. Two-fold increases in BHg and UHg were associated with 0.21 points (95 % confidence interval [CI]: 0.08-0.35) and 0.19 points (95 % CI: 0.09-0.30) higher HSI, respectively. In the WQS model, heavy metal mixture was significantly associated with increased HSI (ß = 0.06, 95 % CI: 0.01-0.11). Similarly, in the BKMR model, heavy metal mixture was positively associated with NAFLD biomarkers, and BHg was the most important contributor in the association. Conclusions: BHg and UHg were significantly associated with NAFLD biomarkers in adolescents, indicating that organic and inorganic mercury exposure could potentially be a risk factor for NAFLD. To mitigate and address the risk of NAFLD associated with heavy metal exposure, it is imperative to take measure to reduce avoidable mercury exposure is necessary.

Cigarette butts enable toxigenic cyanobacteria growth by inhibiting their lethal fungal infections.

Cigarette butts (CBs), of which around 4.5 trillion are discarded annually, are one of the most common types of litter worldwide. CBs contain various chemicals, including metals, nicotine, and polycyclic aromatic hydrocarbons, which can leach into water and pose a threat to aquatic organisms such as cyanobacteria and chytrid fungi. Chytrids, zoosporic fungi that parasitize cyanobacteria lethally, play a crucial role in regulating cyanobacteria blooms by delaying or suppressing bloom formation. Despite the prevalence of CBs in the environment, the impact of their leachates on cyanobacteria-chytrid interactions is not well understood. We assessed the effects of CB leachate on the interaction between the toxigenic cyanobacterium Planktothrix agardhii and its chytrid parasite Rhizophydium megarrhizum. CB leachate inhibited cyanobacterial growth in uninfected cultures. Infection prevalence decreased at 0.2, 2, and 10 CB L-1, with the two highest concentrations completely suppressing infection. Interestingly, at the highest CB concentration, cyanobacterial biomass in infected cultures was comparable to that of uninfected cultures not exposed to CB leachate, suggesting that the presence of chytrids mitigates the impact of the leachate. This study demonstrates that CB leachates are a potential environmental hazard that can enable cyanobacterial growth by inhibiting chytrid infections during epidemics. In addition, our research highlights the importance of assessing the effects of chemical mixtures, such as those leached from CBs, on multi-species interactions, such as host-parasite dynamics. These assessments are crucial to better understand the impact of pollutants on aquatic ecosystems.

Aggregation Behavior of CHR-bis(BODIPY) Bichromophores in THF-water Mixtures: Effect of Linking Positions and Aryl-spacer Substituents.

Aggregation-caused quenching effect (ACQ) greatly limits the practical use of many organic luminophores in biomedicine, optics and electronics. The comparative analysis of aggregation characteristics of CHR-bis(BODIPY) bichromophores 1-6 with R = H, Ph, MeOPh and various linking positions (α,α-; α,ß-; ß,ß- and ß',ß'-) in THF-water mixtures with different water fractions or dye concentrations is first presented in this article. Both the linking style 1-4 and the arylation of the spacer with phenyl (Ph-) 5 or methoxyphenyl (MeOPh-) 6 substituents strongly affect the formation of luminophore aggregated forms in binary THF-water mixtures. The α,α-and ß,ß-isomers (1 and 3) form non-fluorescent H-type aggregates in THF-water mixtures with fw > 70%. The α,ß-; ß',ß'-isomers (2, 4) and the MeOPh-substituted ß,ß-bichromophore 6 are characterized by predominant formation fluorescent aggregates. All bichromophores are characterized by the presence of residual amounts of non-aggregated forms in binary mixtures with maximum water content. The results are useful for controlling the aggregation behavior and spectral characteristics of CHR-bis(BODIPY) bichromophores in aqueous-organic media, which is important in the development of biomarkers and PDT agents.

Semi-quantitative chemometric models for characterization of mixtures of sugars using infrared spectral data.

Sugars (saccharides) are sweet-tasting carbohydrates that are abundant in foods and play very important roles in living organisms, particularly as sources and stores of energy, and as structural elements in cellular membranes. They are desirable therapeutic targets, as they participate in multiple metabolic processes as fundamental elements. However, the physicochemical characterization of sugars is a challenging task, mostly due to the structural similarity shared by the large diversity of compounds of this family. The need for fast, accurate enough, and cost-effective analytical methods for these substances is of extreme relevance, in particular because of the recently increasing importance of carbohydrates in Medicine and food industry. With this in view, this work focused on the development of chemometric models for semi-quantitative analysis of samples of different types of sugars (glucose, galactose, mannitol, sorbose and fructose) using infrared spectra as data, as an example of application of a novel approach, where the Principal Component Analysis (PCA) score plots are used to estimate the composition (weight-%) of the mixtures of the sugars. In these plots, polygonal geometric shapes emerge in the vectorial space of the most significant principal components, that allow grouping different types of samples on the vertices, edges, faces and interior of the polygons according to the composition of the samples. This approach was applied successfully to mixtures of up to 5 sugars and shown to appropriately extract the compositional information from the hyper-redundant complex spectral data. Thought the method has been applied here to a specific problem, it shall be considered as a general procedure for the semi-quantitative analysis of other types of mixtures and applicable to other types of data reflecting their composition. In fact, the methodology appears as an efficient tool to solve three main general problems: (i) use hyper-redundant (in variables) data, as spectral information, directly and with minimum pre-treatment, to evaluate semi-quantitatively the composition of mixtures; (ii) do this for systems which produce data that can be considered rather similar; and (iii) do it for a number of substances present in the mixtures that might be greater than that usually considered in chemistry, which in general is limited to 3 components. In addition, this work also demonstrates that, similarly to the developed analysis based on the PCA score plots, the Multivariate Curve Resolution with Alternating Least Squares (MCR-ALS) chemometric method can also be used successfully for the qualitative (when used without any previous knowledge of the components present in the samples) or semi-quantitative (when the pure components spectral profiles are provided as references) analyses of mixtures of (at least) up to 5 distinct sugars.

Mixed contaminant exposure in tapwater and the potential implications for human-health in disadvantaged communities in California.

Water is an increasingly precious resource in California as years of drought, climate change, pollution, as well as an expanding population have all stressed the state's drinking water supplies. Currently, there are increasing concerns about whether regulated and unregulated contaminants in drinking water are linked to a variety of human-health outcomes particularly in socially disadvantaged communities with a history of health risks. To begin to address this data gap by broadly assessing contaminant mixture exposures, the current study was designed to collect tapwater samples from communities in Gold Country, the San Francisco Bay Area, two regions of the Central Valley (Merced/Fresno and Kern counties), and southeast Los Angeles for 251 organic chemicals and 32 inorganic constituents. Sampling prioritized low-income areas with suspected water quality challenges and elevated breast cancer rates. Results indicated that mixtures of regulated and unregulated contaminants were observed frequently in tapwater throughout the areas studied and the types and concentrations of detected contaminants varied by region, drinking-water source, and size of the public water system. Multiple exceedances of enforceable maximum contaminant level(s) (MCL), non-enforceable MCL goal(s) (MCLG), and other health advisories combined with frequent exceedances of benchmark-based hazard indices were also observed in samples collected in all five of the study regions. Given the current focus on improving water quality in socially disadvantaged communities, our study highlights the importance of assessing mixed-contaminant exposures in drinking water at the point of consumption to adequately address human-health concerns (e.g., breast cancer risk). Data from this pilot study provide a foundation for future studies across a greater number of communities in California to assess potential linkages between breast cancer rates and tapwater contaminants.

Mix and manage: Cultivar mixtures can maintain yield under high wheat blast disease pressure.

Originating in South America, wheat blast disease has spread to both Asia and Africa and is considered a significant threat to food security. Bangladesh experienced the first outbreak of wheat blast outside of the Americas in 2016. Shortly thereafter, the blast-resistant variety BARI Gom 33 was released. Seeds of this variety are however not as widely available as required, although the disease threat remains. While varietal mixtures have been shown to mitigate some symptoms and yield losses associated with other fungal diseases in wheat, there is a complete research gap on this topic as it pertains to wheat blast. As such, we evaluated the potential of using BARI Gom 33 as a component of a variety mixture under high disease pressure in Bangladesh. During three cropping seasons, blast symptoms and yield were determined in a field experiment for the highly blast-susceptible variety BARI Gom 26, the moderately susceptible BARI Gom 30, the resistant BARI Gom 33, and seven mixture combinations of the three varieties using artificial inoculation to increase disease pressure. In addition to wheat blast, Bipolaris leaf blight (BpLB) symptoms were observed and evaluated. While yields of the susceptible varieties were severely affected by blast even after fungicide application, disease-inflicted yield loss without fungicide was only 15% for sole BARI Gom 33 and did not differ significantly from yield losses in BARI Gom 33 and BARI Gom 30 mixtures. Furthermore, in the mixture containing 67% BARI Gom 33 and 33% BARI Gom 30, blast incidence and severity were reduced by 25% and 16%, respectively, in comparison to weighted values in sole stands. Conversely, mixing varieties tended to increase the symptoms of BpLB. Under high wheat blast pressure, fungicide protection against blast was relatively weak, underscoring the importance of resistant varieties. Although variety mixtures did not increase yield, the yield advantage of BARI Gom 33 was maintained when its seeds were mixed with the less resistant BARI Gom 30. This study confirms recommendations that farmers should use BARI Gom 33 as a first line of defense against wheat blast in Bangladesh. Yet where farmers cannot access sufficient BARI Gom 33 seed for planting, our data suggest that agricultural extension services can recommend this variety with non-resistant cultivars as interim strategy without significant risk of yield loss.

Cocktail effects of clothianidin and imidacloprid in zebrafish embryonic development, with high and low concentrations of mixtures.

There is growing concern that sprayed neonicotinoid pesticides (neonics) persist in mixed forms in the environmental soil and water systems, and these concerns stem from reports of increase in both the detection frequency and concentration of these pollutants. To confirm the toxic effects of neonics, we conducted toxicity tests on two neonics, clothianidin (CLO) and imidacloprid (IMD), in embryos of zebrafish. Toxicity tests were performed with two different types of mixtures: potential mixture compounds and realistic mixture compounds. Potential mixtures of CLO and IMD exhibited synergistic effects, in a dose-dependent manner, in zebrafish embryonic toxicity. Realistic mixture toxicity tests that are reflecting the toxic effects of mixture in the aquatic environment were conducted with zebrafish embryos. The toxicity of the CLO and IMD mixture at environmentally-relevant concentrations was confirmed by the alteration of the transcriptional levels of target genes, such as cell damage linked to oxidative stress response and thyroid hormone synthesis related to zebrafish embryonic development. Consequently, the findings of this study can be considered a strategy for examining mixture toxicity in the range of detected environmental concentrations. In particular, our results will be useful in explaining the mode of toxic action of chemical mixtures following short-term exposure. Finally, the toxicity information of CLO and IMD mixtures will be applied for the agricultural environment, as a part of chemical regulation guideline for the use and production of pesticides.

Olfactory perception complexity induced by key odorants perceptual interactions of alcoholic beverages: Wine as a focus case example.

The odorants in alcoholic beverages are frequently experienced as complex mixtures, and there is a complex array of influence factors and interactions involved during consumption that deeply increase its olfactory perception complexity, especially the complexity induced by perceptual interactions between different odorants. In this review, the effect of olfactory perceptual interactions and other factors related to the complexity of olfactory perception of alcoholic beverages are discussed. The classification, influencing factors, and mechanisms of olfactory perceptual interactions are outlined. Recent research progress as well as the methodologies applied in these studies on perceptual interactions between odorants observed in representative alcoholic beverages, especially wine, are briefly summarized. In the future, unified theory or systematic research methodology need to be established, since up to now, the rules of perceptual interaction between multiple odorants, which is critical to the alcoholic beverage industry to improve the flavor of their products, are still not revealed.

Assessment of Neurotoxic Mechanisms of Individual and Binary Mixtures of Cobalt, Nickel and Lead in Hippocampal Neuronal Cells.

Many studies have focused on the neurotoxic effects of single metals, while investigation on the exposure to metal mixtures, which mainly occur in real-life situations, is scarce. This study sought to assess the neurotoxic effect of Ni, Co, and Pb binary mixtures and their individual effects in hippocampal neuronal cells (HT-22). Cells were exposed to Ni, Co, and Pb separately for 48 h at 37°C and 5% CO2, and cell viability was assessed. Morphological assessment of the cells exposed to binary mixtures of Co, Ni, and Pb and single metals was assessed using a microscope. Furthermore, acetylcholinesterase (AChE) activity, oxidative stress biomarkers (glutathione [GSH] and malondialdehyde [MDA] levels, catalase [CAT], and glutathione-S transferase [GST] activities) and nitric oxide [NO] levels were evaluated after treatment with the binary mixtures and single metals. Binary mixtures of the metals reduced cell viability, exerting an additivity action. The combinations also exerted synergistic action, as revealed by the combination index. Furthermore, a significant reduction in AChE activity, GSH levels, CAT and GST activities, and high MDA and NO levels were observed in neuronal cells. The additive interactions and synergistic actions of the binary mixtures might contribute to the significant reduction of AChE activity, GSH levels, GST, and CAT activities, and an increase in MDA and NO levels. The findings from this study revealed significant evidence that binary mixtures of Co, Pb, and Ni may induce impaired neuronal function and, ultimately, neurodegeneration.

Solubility and thermodynamic analysis of aceclofenac in different {Carbitol + water} mixtures at various temperatures.

The solubility and thermodynamic properties of the anti-inflammatory drug aceclofenace (ACF) have been assessed in a range of {2-(2-ethoxyethoxy)ethanol (Carbitol) + water} combinations at temperatures ranging from 298.2 K to 318.2 K and atmospheric pressure of 101.1 kPa. The shake flask method was employed to determine the solubility of ACF, and various models including "van't Hoff, Apelblat, Buchowski-Ksiazczak λh, Yalkowsky-Roseman, Jouyban-Acree, and Jouyban-Acree-van't Hoff models" were used to validate the results. The computational models demonstrated a strong correlation with the experimental ACF solubility data, as indicated by the error values of < 3.0%. In the compositions of {Carbitol + water}, the ACF mole fraction solubility was enhanced by temperature and Carbitol mass fraction. The solubility of ACF in mole fraction was found to be lowest in pure water (1.07 × 10- 6 at 298.2 K), and highest in pure Carbitol (1.04 × 10- 1 at 318.2 K). Based on the positive values of the calculated thermodynamic parameters, the dissolution of ACF was determined to be "endothermic and entropy-driven" in all of the {Carbitol + water} solutions that were studied. It was also observed that enthalpy controls the solvation of ACF in solutions containing {Carbitol + water}. ACF-Carbitol had the strongest molecular interactions in contrast to ACF-water. Based on the results of this study, Carbitol holds significant potential for enhancing the solubility of ACF in water.

Hyperpolarized 13C NMR Reveals Pathway Regulation in Lactococcus lactis and Metabolic Similarities and Differences Across the Tree of Life.

The control of metabolic networks is incompletely understood, even for glycolysis in highly studied model organisms. Direct real-time observations of metabolic pathways can be achieved in cellular systems with 13C NMR using dissolution Dynamic Nuclear Polarization (dDNP NMR). The method relies on a short-lived boost of NMR sensitivity using a redistribution of nuclear spin states to increase the alignment of the magnetic moments by more than four orders of magnitude. This temporary boost in sensitivity allows detection of metabolism with sub-second time resolution. Here, we hypothesized that dDNP NMR would be able to investigate molecular phenotypes that are not easily accessible with more conventional methods. The use of dDNP NMR allows real-time insight into carbohydrate metabolism in a Gram-positive bacterium (Lactoccocus lactis), and comparison to other bacterial, yeast and mammalian cells shows differences in the kinetic barriers of glycolysis across the kingdoms of life. Nevertheless, the accumulation of non-toxic precursors for biomass at kinetic barriers is found to be shared across the kingdoms of life. We further find that the visualization of glycolysis using dDNP NMR reveals kinetic characteristics in transgenic strains that are not evident when monitoring the overall glycolytic rate only. Finally, dDNP NMR reveals that resting Lactococcus lactis cells use the influx of carbohydrate substrate to produce acetoin rather than lactate during the start of glycolysis. This metabolic regime can be emulated using suitably designed substrate mixtures to enhance the formation of the C4 product acetoin more than 400-fold. Overall, we find that dDNP NMR provides analytical capabilities that may help to clarify the intertwined mechanistic determinants of metabolism and the optimal usage of biotechnologically important bacteria.

The potential of human leukocyte antigen alleles to assist with multiple-contributor DNA mixtures: Proof of concept study.

One of the most challenging issues still present in forensic DNA analysis is identifying individuals in samples containing DNA from multiple contributors. The introduction of novel identification markers may be a useful tool in the deconvolution of such DNA mixtures. In this study, we investigated the potential of alleles from the human leukocyte antigen system (HLA) to aid in identifying individuals in complex, multiple-donor DNA samples. The most advantageous characteristic of the HLA complex is its polymorphism in the human genome. A 22-loci multiplex with HLA markers was designed and applied to two-, three-, and four-person DNA mixtures. The results of the conducted experiments demonstrated that the identification of individuals in multiple contributor samples with the help of HLA markers is possible; however, it is clear that the reliability of the method is heavily dependent on the number of unique alleles for each individual in the analysed mixture. In order to compare this novel approach against the already established process, the same group of reference and multiple-contributor samples was analysed with a commonly used set of STR markers. This proof-of-concept research shows the importance of examining alternative solutions to the current deconvolution challenge in forensic DNA profiling.

Predictive toxicology of chemical mixtures using proteome-wide thermal profiling and protein target properties.

Our capability to predict the impact of exposure to chemical mixtures on environmental and human health is limited in comparison to the advances on the chemical characterization of the exposome. Current approaches, such as new approach methodologies, rely on the characterization of the chemicals and the available toxicological knowledge of individual compounds. In this study, we show a new methodological approach for the assessment of chemical mixtures based on a proteome-wide identification of the protein targets and revealing the relevance of new targets based on their role in the cellular function. We applied a proteome integral solubility alteration assay to identify 24 protein targets from a chemical mixture of 2,3,7,8-tetrachlorodibenzo-p-dioxin, alpha-endosulfan, and bisphenol A among the HepG2 soluble proteome, and validated the chemical mixture-target interaction orthogonally. To define the range of interactive capability of the new targets, the data from intrinsic properties of the targets were retrieved. Introducing the target properties as criteria for a multi-criteria decision-making analysis called the analytical hierarchy process, the prioritization of targets was based on their involvement in multiple pathways. This methodological approach that we present here opens a more realistic and achievable scenario to address the impact of complex and uncharacterized chemical mixtures in biological systems.

Applications of mixture methods in epidemiological studies investigating the health impact of persistent organic pollutants exposures: a scoping review.

BACKGROUND: Persistent organic pollutants (POPs) are environmental chemicals characterized by long half-lives in nature and human bodies, posing significant health risks. The concept of the exposome, encompassing all lifetime environmental exposures, underscores the importance of studying POP as mixtures rather than in isolation. The increasing body of evidence on the health impacts of POP mixtures necessitates the proper application of statistical methods. OBJECTIVES: We aimed to summarize studies on the overall effects of POP mixtures, identify patterns in applications of mixture methods-statistical methods for investigating the association of mixtures-and highlight current challenges in synthesizing epidemiologic evidence of POP mixtures on health effects as illustrated through a case study. METHODS: We conducted a systematic literature search on PubMed and Embase for epidemiological studies published between January 2011 and April 2023. RESULTS: We included 240 studies that met our eligibility criteria. 126 studies focused on per- and polyfluoroalkyl substances (PFAS) mixtures only, while 40 analyzed three or more classes of POPs in mixture analyses. We identified 23 unique mixture methods used to estimate the overall effects of POP mixtures, with Bayesian Kernel Machine Regression (BKMR), a type of response-surface modeling, being the most common. Additionally, 22.9% of studies used a combination of methods, including response-surface modeling, index modeling, dimension reduction, and latent variable models. The most extensively explored health outcome category was body weight and birth sizes (n = 43), and neurological outcomes (n = 41). In the case study of PFAS mixtures and birth weight, 12 studies showed negative associations, while 4 showed null results, and 2 showed positive associations. IMPACT STATEMENT: This scoping review consolidates the existing literature on the overall effects of POP mixtures using statistical methods. By providing a comprehensive overview, our study illuminates the present landscape of knowledge in this field and underscores the methodological hurdles prevalent in epidemiological studies focused on POP mixtures. Through this analysis, we aim to steer future research directions, fostering a more nuanced comprehension of the intricate dynamics involved in assessing the health effects of POP mixtures. Our work stands as a significant contribution to the ongoing exploration of the chemical exposome.

Effects of microplastic interaction with persistent organic pollutants on the activity of the aryl hydrocarbon and estrogen receptors.

Environmental microplastics (MPs) are complex mixtures of plastic polymers and sorbed chemical pollutants with high degrees of heterogeneity, particularly in terms of particle size, morphology and degree of weathering. Currently, limitations exist in sampling sufficient amounts of environmental particles for laboratory studies to assess toxicity endpoints with statistical rigor and to examine chemical pollutant interactions. This study seeks to bridge this gap by investigating environmental plastic particle mimetics and pollutant-polymer interactions by mixing polymer particles with persistent organic pollutants (POPs) at set concentrations over time. Solutions containing combinations of polymers including polystyrene (PS), polypropylene (PP), polyethylene terephthalate (PET), and polyamide (PA) and POPs including 2,3,7,8 -Tetrachlorodibenzo-p-dioxin (TCDD), bisphenol A (BPA), and atrazine, were stirred for up to 19 weeks and monitored using assays to test for aryl hydrocarbon (AhR) and estrogen receptor (ER) activity which are cell signaling pathways impacted by environmental pollutants. TCDD induced AhR activity decreased over time in the presence of PS in a surface area dependent manner. BPA and atrazine also exhibited AhR antagonist activity in the presence of TCDD. The addition of BPA slowed the loss of activity but atrazine did not, suggesting that polymer chemistry impacts interactions with POPs. We also observed potential differences in TCDD sorption with different plastic polymers and that higher concentrations of PS particles may inhibit BPA-induced estrogen receptor activation. These results emphasize the need for additional understanding of how POPs and polymer chemistry impact their interaction and toxicity.

Thermo-responsible PNIPAM-grafted polystyrene microspheres for mesenchymal stem cells culture and detachment.

The preparation of cells is a critical step in cell therapy. To ensure the effectiveness of cells used for clinical treatments, it is essential to harvest adherent cells from the culture media in a way that preserves their high viability and full functionality. In this study, we developed temperature-responsive poly(N-isopropylacrylamide) (PNIPAM)-grafted polystyrene (PS) microspheres using reversible addition-fragmentation chain transfer polymerization. These microspheres allow for the non-destructive harvesting of cultured cells through temperature changes. The composition and physicochemical properties of the PNIPAM-grafted PS microspheres were confirmed using infrared spectroscopy, elemental analysis, dynamic light scattering, and thermogravimetric analysis.In vitroexperiments demonstrated that these microspheres exhibit excellent biocompatibility, supporting the adhesion and proliferation of various cells. Moreover, the microspheres showed good temperature responsiveness in thermosensitive detachment experiments with GFP-HepG2cells and umbilical cord mesenchymal stem cells (UC-MSCs). Additionally, through orthogonal experiments, we identified a cell detachment aid mixture that significantly improved the dispersibility of cells detached from the microspheres, enhancing the efficiency of thermosensitive cell detachment by approximately 40%. The harvested UC-MSCs retained their capacity for re-proliferation and trilineage differentiation. Consequently, the temperature-responsive microspheres developed in this study, combined with the cell detachment aid mixtures, hold great potential for large-scale culture and harvesting of therapeutic cells in clinical applications.

Entropy Analysis of Implicit Heat Fluxes in Multi-Temperature Mixtures.

We propose new implicit constitutive relations for the heat fluxes of a two-temperature mixture of fluids. These relations are frame-indifferent forms. However, classical explicit forms of the stress tensors and the interaction forces (specified as explicit forms of constitutive relations) as given in mixture theory are used. The focus here is to establish constraints imposed on the implicit terms in the heat fluxes due to the Second Law of Thermodynamics. Our analysis establishes that the magnitude of the explicit entropy production is equal to or greater than that of the implicit entropy production.

Combinatory Effects of Acrylamide and Deoxynivalenol on In Vitro Cell Viability and Cytochrome P450 Enzymes of Human HepaRG Cells.

Acrylamide (AA) can be formed during the thermal processing of carbohydrate-rich foods. Deoxynivalenol (DON), a mycotoxin produced by Fusarium spp., contaminates many cereal-based products. In addition to potential co-exposure through a mixed diet, co-occurrence of AA and DON in thermally processed cereal-based products is also likely, posing the question of combinatory toxicological effects. In the present study, the effects of AA (0.001-3 mM) and DON (0.1-30 µM) on the cytotoxicity, gene transcription, and expression of major cytochrome P450 (CYP) enzymes were investigated in differentiated human hepatic HepaRG cells. In the chosen ratios of AA-DON (10:1; 100:1), cytotoxicity was clearly driven by DON and no overadditive effects were observed. Using quantitative real-time PCR, about twofold enhanced transcript levels of CYP1A1 were observed at low DON concentrations (0.3 and 1 µM), reflected by an increase in CYP1A activity in the EROD assay. In contrast, CYP2E1 and CYP3A4 gene transcription decreased in a concentration-dependent manner after incubation with DON (0.01-0.3 µM). Nevertheless, confocal microscopy showed comparably constant protein levels. The present study provided no indication of an induction of CYP2E1 as a critical step in AA bioactivation by co-occurrence with DON. Taken together, the combination of AA and DON showed no clear physiologically relevant interaction in HepaRG cells.

Elementomics of 32 elements in cord serum depicts the risk of orofacial clefts: A case-control study in Shanxi, China.

Maternal exposure to various metallic and non-metallic elements has been linked to the occurrence of orofacial clefts (OFCs), yet there remains a dearth of comprehensive research on the potential ramifications of simultaneous exposure to multiple elements. In this study, we investigated the individual and combined effects of element exposure on OFCs in a cohort of 168 pregnant women (49 cases and 119 controls) in the Shanxi province of northern China from 2010 to 2015. Cord serum samples were obtained from all participants to analyze the levels of 32 elements using inductively coupled plasma-mass spectrometry. The study examined the independent correlation between element concentrations and OFCs using two machine screening models, Boruta and Least Absolute Shrinkage and Selection Operator. Bayesian kernel machine regression (BKMR) was utilized to determine the combined effects of key exposure elements on OFCs and to clarify the interaction between exposed elements through the generalized additive model (GAM). The screening models identified lead (Pb), tin (Sn), iron (Fe), and cesium (Cs) as the most significant risk factors for OFC development in offspring. In the BKMR model, the probability of OFCs increased with higher overall levels of these risk elements, with Pb emerging as the primary contributor to the combined effect of the mixture. The findings of the GAM indicated that the combined exposure to Pb and Sn had a synergistic effect on the risk of developing OFCs. Analysis of elemental exposure in umbilical cord serum suggested that Pb exposure may have detrimental effects on OFC development in offspring, which may be further intensified by a synergistic interaction between Sn and Pb in the occurrence of OFCs.

17α-Ethynylestradiol and Levonorgestrel Exposure of Rainbow Trout RTL-W1 Cells at 18 °C and 21 °C Mainly Reveals Thermal Tolerance, Absence of Estrogenic Effects, and Progestin-Induced Upregulation of Detoxification Genes.

Fish are exposed to increased water temperatures and aquatic pollutants, including endocrine-disrupting compounds (EDCs). Although each stressor can disturb fish liver metabolism independently, combined effects may exist. To unveil the molecular mechanisms behind the effects of EDCs and temperature, fish liver cell lines are potential models needing better characterisation. Accordingly, we exposed the rainbow trout RTL-W1 cells (72 h), at 18 °C and 21 °C, to ethynylestradiol (EE2), levonorgestrel (LNG), and a mixture of both hormones (MIX) at 10 µM. The gene expression of a selection of targets related to detoxification (CYP1A, CYP3A27, GST, UGT, CAT, and MRP2), estrogen exposure (ERα, VtgA), lipid metabolism (FAS, FABP1, FATP1), and temperature stress (HSP70b) was analysed by RT-qPCR. GST expression was higher after LNG exposure at 21 °C than at 18 °C. LNG further enhanced the expression of CAT, while both LNG and MIX increased the expressions of CYP3A27 and MRP2. In contrast, FAS expression only increased in MIX, compared to the control. ERα, VtgA, UGT, CYP1A, HSP70b, FABP1, and FATP1 expressions were not influenced by the temperature or the tested EDCs. The RTL-W1 model was unresponsive to EE2 alone, sensitive to LNG (in detoxification pathway genes), and mainly insensitive to the temperature range but had the potential to unveil specific interactions.