Ligand Control of Copper-Mediated Cycloadditions of Acetylene to Azides: Chemo- and Regio-Selective Formation of Deutero- and Iodo-Substituted 1,2,3-Triazoles.

The participation of σ-monocopper and σ-bis-copper acetylide in mechanistic pathways for copper-catalyzed cycloaddition (CuAAC) reactions of acetylene with azides was probed by analysis of deuterium distributions in the 1,2,3-triazole product formed by deuterolysis of initially formed mono- and bis-copper triazoles. The results show that, when Cu(Phen)(PPh3)2NO3 is used as the catalyst for reactions of acetylene with azides in DMF/D2O, 1-substituted-5-deutero-1,2,3-triazoles are generated selectively. This finding demonstrates that the Cu(Phen)(PPh3)2NO3-catalyzed cycloadditions utilize monocopper acetylide as the substrate and produce 5-copper-1,2,3-triazoles initially. Conversely, when DBU or Et3N is the copper ligand, the process takes place through initial formation and cycloaddition of bis-copper acetylide to produce 4,5-bis-copper-triazole, which reacts with D2O to form the corresponding 4,5-bis-deutero-triazole. Moreover, when C2D2 is used as the substrate, Cu(Phen)(PPh3)2NO3 as the Cu ligand, and H2O/DMF as the solvent, mono-C4-deutreo 1,2,3-triazoles are generated in high yields and excellent levels of regioselectivity. Lastly, CuAAC reactions of acetylene with azides, promoted by CuCl2·2H2O and NaI, yield 4,5-diiodo-1,2,3-triazoles with moderate to high efficiencies.

A framework enabling LLMs into regulatory environment for transparency and trustworthiness and its application to drug labeling document.

Regulatory agencies consistently deal with extensive document reviews, ranging from product submissions to both internal and external communications. Large Language Models (LLMs) like ChatGPT can be invaluable tools for these tasks, however present several challenges, particularly the proprietary information, combining customized function with specific review needs, and transparency and explainability of the model's output. Hence, a localized and customized solution is imperative. To tackle these challenges, we formulated a framework named askFDALabel on FDA drug labeling documents that is a crucial resource in the FDA drug review process. AskFDALabel operates within a secure IT environment and comprises two key modules: a semantic search and a Q&A/text-generation module. The Module S built on word embeddings to enable comprehensive semantic queries within labeling documents. The Module T utilizes a tuned LLM to generate responses based on references from Module S. As the result, our framework enabled small LLMs to perform comparably to ChatGPT with as a computationally inexpensive solution for regulatory application. To conclude, through AskFDALabel, we have showcased a pathway that harnesses LLMs to support agency operations within a secure environment, offering tailored functions for the needs of regulatory research.

Exploring the association between campus environment of higher education and student health: A systematic review of findings and measures.

Numerous studies have investigated the relationship between the neighborhood environments and residents' health. However, other important settings, such as university campuses, have received little attention. This paper conducted a systematic review and synthesized existing empirical works examining the association between the university/college campuses built and natural environments and students' health. Following the PRISMA guidelines, we searched nine databases using keywords related to higher-education campuses and health-related outcomes. A total of 19 articles were identified, including fifteen cross-sectional studies, three experimental studies, and one longitudinal study. The majority of the studies were conducted in Asian countries and published in the past five years. The findings indicate that active transportation infrastructure, such as increased road intersections and better walkability, were found to be positively associated with students' physical activity. The natural environments, including perceived naturalness, blue space, and greenness was shown to support student's mental health and quality of life. Specifically, blue space was found to be the most preferred place for mental restoration, and scattered trees demonstrated a supportive effect in reducing depression symptoms. Even just viewing virtual trees had a restorative effect and feel less anxiety. Additionally, during the summer, tree shadows were identified as the most important factors for enhancing thermal comfort. This review emphasizes the crucial role of campus environments in promoting college students' health. Future longitudinal studies and investigations using multiple campuses would provide a more comprehensive understanding of this relationship. Such endeavors can contribute to the development of evidence-based strategies for designing and planning healthy campus environments that optimize students' well-being.

The spatial relationship between long-term vacant housing and non-communicable diseases in U.S. shrinking and growing metropolitan areas.

The rising prevalence of non-communicable diseases (NCDs) has led to increased attention on understanding how built environments affect NCD risks. However, there's a significant gap in the literature regarding the relationship between housing vacancy duration and NCDs in metropolitan areas with varying development rates. Our research addresses this gap by examining the association between housing vacancy duration and NCDs across all U.S. metropolitan areas, considering growing, shrinking, and fluctuating counties. We used a Multiscale Geographically Weighted Regression (MGWR) model to analyze this relationship, finding that longer-term vacant housing (over 3 years) is more positively associated with NCDs compared to short-term vacancies. We also discovered that this association is non-uniform across metropolitan counties, except for cancer and stroke outcomes. Shrinking counties in the Northeast are particularly affected, emphasizing the need for targeted public health interventions in these areas. This study underscores the importance of revitalizing vacant homes, especially those vacant for over 3 years, in both shrinking and growing regions to improve public health. Policymakers should adopt tailored strategies, engage public health experts, and invest in healthcare infrastructure to effectively address the health risks linked to vacant housing.

Dynamic evaluation of pathological changes in a mouse acne model by optical imaging technology.

Acne vulgaris is a disorder of the pilosebaceous unit that is primarily caused by hyperseborrhoea, colonization with Propionibacterium acnes, hyperkeratosis and an inflammatory response. Existing pharmacodynamic assessment methods primarily focus on a single causative factor at a certain time point, making it difficult to assess multiple factors simultaneously in real time. Therefore, it is crucial to establish a dynamic and nondestructive method for the assessment of acne in vivo. This study utilized four-dimensional optical imaging techniques to assess the pathogenic factors and pathological progression of acne. LSCI was employed to measure blood flow; TPEF was used to observe inflammatory changes (NAD(P)H) in epidermal granular layer cells and structural changes in collagen fibres in the dermal layer. Additionally, the dermatoscope was used to investigate the micro-characterization of the lesions. We observed that the epidermis in the lesion area was thickened, hair follicles were keratinized, and there was obvious inflammation and blood flow aggregation by optical imaging technology. Based on these findings, the pathological progression of this acne model could be divided into the inflammation phase, accompanied by bacterial colonization, and the reparative phase. These results provide a new perspective for the assessment of acne and offer an experimental basis for the selection of precise drugs for clinical use.

The charge effects on the hydrogen evolution reaction activity of the defected monolayer MoS2.

Doping engineering has proven to be an effective way to tune the hydrogen evolution reaction (HER) activity of MoS2. Introducing these defects could cause the overall charge imbalance of MoS2, which makes MoS2 charged. In order to understand the effect of charge on the HER activity of the defected MoS2, we systematically investigate the formation energies, hydrogen adsorption Gibbs free energy (), and electronic structures of 3d, 4d, and 5d transition metal (TM) doped monolayer MoS2 with S vacancies (Svac) based on the density functional theory (DFT) calculations. According to the formation energy calculation, Svac in the 0 and -1 charge states (S0vac and Svac1-) is found to be stable. of Svac1- is -0.16 eV, suggesting its HER catalytic activity is lower than that of Pt (), which is consistent with the experimental results. By substituting the Mo atom with TM atoms, we found that the TM atoms in groups VB-VIIB can promote the generation of Svac, forming defect complexes (TMMoSvac). is greatly affected by the charge state of defects; TMMoSvac defects (TM = V, Nb, Ta, Cr, W, Mn, and Re) in -1 charge states exhibited excellent HER activity (). Significantly, W and Re doping can promote the HER activity of MoS2 independent of the charge state and the Fermi level, which suggests that W and Re doping are most beneficial to improve the HER activity of MoS2. Therefore, the HER activity of defected MoS2 is not only influenced by as previously thought, but also by formation energies, charge state and Fermi level position of defects. The underlying physics could be deduced from the charge-induced changes in electronic structures. Our work highlights the defect charge effects on the electrochemical reactions and offers plausible mechanisms of defect charge effects.

Impact of heat on all-cause and cause-specific mortality: A multi-city study in Texas.

BACKGROUND: Studies on the health effects of heat are particularly limited in Texas, a U.S. state in the top 10 highest number of annual heat-related deaths per capita from 2018 to 2020. This study assessed the effects of heat on all-cause and cause-specific mortality in 12 metropolitan statistical areas (MSAs) across Texas from 1990 to 2011. METHODS: First, we determined the heat thresholds for each MSA above which the relation between temperature and mortality is linear. We then conducted a distributed lag non-linear model for each MSA, followed by a random effects meta-analysis to estimate the pooled effects for all MSAs. We repeated this process for each mortality cause and age group to achieve the effect estimates. RESULTS: We found a 1 °C temperature increase above the heat threshold is associated with an increase in the relative risk of all-cause mortality of 0.60% (95%CI [0.39%, 0.82%]) and 1.10% (95%CI [0.65%, 1.56%]) for adults older than 75. For each MSA, the relative risk of mortality for a 1 °C temperature increase above the heat threshold ranges from 0.10% (95%CI [0.09%, 0.10%]) to 1.29% (95%CI [1.26%, 1.32%]). Moreover, elevated temperatures showed a slight decrease in cardiovascular mortality (0.37%, 95%CI [-0.35%, 1.09%]) and respiratory disease (1.97%, 95%CI [-0.11%, 4.08%]), however this effect was not considered statistically significant.. CONCLUSION: Our study found that high temperatures can significantly impact all-cause mortality in Texas, and effect estimates differ by MSA, age group, and cause of death. Our findings generate critical information on the impact of heat on mortality in Texas, providing insights for policymakers on resource allocation and strategic intervention to reduce heat-related health effects.

Applying genomics in regulatory toxicology: a report of the ECETOC workshop on omics threshold on non-adversity.

In a joint effort involving scientists from academia, industry and regulatory agencies, ECETOC's activities in Omics have led to conceptual proposals for: (1) A framework that assures data quality for reporting and inclusion of Omics data in regulatory assessments; and (2) an approach to robustly quantify these data, prior to interpretation for regulatory use. In continuation of these activities this workshop explored and identified areas of need to facilitate robust interpretation of such data in the context of deriving points of departure (POD) for risk assessment and determining an adverse change from normal variation. ECETOC was amongst the first to systematically explore the application of Omics methods, now incorporated into the group of methods known as New Approach Methodologies (NAMs), to regulatory toxicology. This support has been in the form of both projects (primarily with CEFIC/LRI) and workshops. Outputs have led to projects included in the workplan of the Extended Advisory Group on Molecular Screening and Toxicogenomics (EAGMST) group of the Organisation for Economic Co-operation and Development (OECD) and to the drafting of OECD Guidance Documents for Omics data reporting, with potentially more to follow on data transformation and interpretation. The current workshop was the last in a series of technical methods development workshops, with a sub-focus on the derivation of a POD from Omics data. Workshop presentations demonstrated that Omics data developed within robust frameworks for both scientific data generation and analysis can be used to derive a POD. The issue of noise in the data was discussed as an important consideration for identifying robust Omics changes and deriving a POD. Such variability or "noise" can comprise technical or biological variation within a dataset and should clearly be distinguished from homeostatic responses. Adverse outcome pathways (AOPs) were considered a useful framework on which to assemble Omics methods, and a number of case examples were presented in illustration of this point. What is apparent is that high dimension data will always be subject to varying processing pipelines and hence interpretation, depending on the context they are used in. Yet, they can provide valuable input for regulatory toxicology, with the pre-condition being robust methods for the collection and processing of data together with a comprehensive description how the data were interpreted, and conclusions reached.

Associations between vacant urban lands and public health outcomes in growing and shrinking cities.

Vacant urban land, although not officially designated as a green space, often exhibits a semi-wild natural state due to being left open to colonization by nature. Attention to the effects of vacant urban land on human health has increased due to both rising urban vacancy rates and non-communicable diseases (NCDs). However, relationships between many vacant land characteristics (such as vegetation coverage, size, duration, and location) and NCDs have not been comprehensively examined, especially comparing shrinking (depopulating) and growing (populating) cities. This study evaluates St. Louis, MO (shrinking), and Los Angeles, CA (growing) to explore these relationships using ordinary least squares (OLS) interaction analysis with a moderator approach. Results show that associations between vacancy rate, duration, location, and NCDs differ significantly between city types. Vegetation coverage and size are associated with specific NCDs, but there are no differences between city types. Unlike the largely dilapidated vacant lands in the shrinking city, which tend to harm public health, vacant lots in the growing city were more functional green spaces that can, in some cases, even mitigate NCDs. Interestingly, In St. Louis, the shorter the average duration of the vacant land, the greater the risk of NCDs in a shrinking city. This is because vacant land can be contagious to nearby lots if not treated, leading to more newly emerged vacant lands and reducing the average duration of vacant land. In such cases, census tracts with the lower duration of vacant lands in St. Louis tend to be areas facing persistent environmental degradation and high public health threats. Regarding location, vacant lands near industrial areas were linked to negative health outcomes in the Los Angeles (growing), while those near single-family and commercial areas posed higher risks of NCDs in the St Louis (shrinking). The findings aid decision-making for land supply regulation and regeneration as well as urban green space management to promote human health and well-being.

Epigenetics in drug disposition & drug therapy: symposium report of the 24th North American meeting of the International Society for the Study of Xenobiotics (ISSX).

The 24th North American International Society for the Study of Xenobiotics (ISSX) meeting, held virtually from September 13 to 17, 2021, embraced the theme of "Broadening Our Horizons." This reinforces a key mission of ISSX: striving to share innovative science related to drug discovery and development. Session speakers and the ISSX New Investigators Group, which supports the scientific and professional development of student and early career ISSX members, elected to highlight the scientific content presented during the captivating session titled, "Epigenetics in Drug Disposition & Drug Therapy." The impact genetic variation has on drug response is well established; however, this session underscored the importance of investigating the role of epigenetics in drug disposition and drug discovery. Session speakers, Drs. Ning, McClay, and Lazarus, detailed mechanisms by which epigenetic players including long non-coding RNA (lncRNAs), microRNA (miRNAs), DNA methylation, and histone acetylation can alter the expression of genes involved in pharmacokinetics, pharmacodynamics, and toxicity. Dr. Ning detailed current knowledge about miRNAs and lncRNAs and the mechanisms by which they can affect the expression of drug metabolizing enzymes (DMEs) and nuclear receptors. Dr. Lazarus discussed the potential role of miRNAs on UDP-glucuronosyltransferase (UGT) expression and activity. Dr. McClay provided evidence that aging alters methylation and acetylation of DMEs in the liver, affecting gene expression and activity. These topics, compiled by the symposium organizers, presenters, and the ISSX New Investigators Group, are herein discussed, along with exciting future perspectives for epigenetics in drug disposition and drug discovery research.

Scattering response modeling scheme based on combined neural network inspired by the equivalent scattering center.

A novel scheme is proposed in this paper to model the complex scattering pattern of radar target with a small training data set. By employing the ideal equivalent scattering center as transfer function, the frequency domain response can be represented by series of parameters so that the aspect and frequency domain dependency can be decoupled, and modeled, independently. In specific, neural network is employed to model the aspect dependency considering the complexity. To maintain the continuity of transformed parameters, a parameter extraction algorithm based on the Orthogonal Matching Pursuit is designed. With the same amount of training set, the proposed scheme exhibits a much better performance than the existing representative modeling techniques such as Geometrical Theory of Diffraction (GTD)-based model, the polynomial scattering center model and so on. At the same time, the training speed of the proposed model is also faster than those techniques.

ATF2 accelerates the invasion and metastasis of hepatocellular carcinoma through targeting the miR-548p/TUFT1 axis.

AIM: Due to high invasion and metastasis, hepatocellular carcinoma (HCC) is known as one of the most fatal carcinomas. We aim to further investigate regulatory mechanisms of invasion and metastasis to elucidate HCC pathogenesis and develop novel medications. METHODS: Patient specimens were collected for assessing gene expression and correlation between gene expressions. The expression of Ki67 and E-cadherin in subcutaneous xenograft tumor were examined by immunohistochemistry staining. The expression of activating transcription factor 2 (ATF2), miR-548p and TUFT1 were determined using Real-time quantitative reverse transcription polymerase chain reaction. Epithelial-mesenchymal transition and PI3K/AKT signaling-associated markers were examined with western blot. The proliferation, migration and invasion were assessed by 3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide, colony formation and transwell assays, respectively. Cell apoptosis was assessed via Annexin V and propidium iodide staining. Gene interaction was confirmed using chromatin immunoprecipitation and luciferase activity assays. Subcutaneous and intravenous xenograft mouse models were established for analyzing HCC growth and metastasis in vivo. RESULTS: ATF2 was up-regulated in HCC patients and cells. ATF2 promoted HCC cell proliferation, migration and invasion and inhibited cell apoptosis through directly targeting miR-548p and controlling its expression. miR-548p suppressed HCC cell proliferation, migration and invasion and enhanced cell apoptosis. miR-548p directly bound to the 3'UTR of TUFT1 to restrain its expression and subsequently suppress the PI3K/AKT signaling. ATF2 knock-down significantly suppressed the growth and metastasis of HCC. CONCLUSION: ATF2 accelerates HCC progression by promoting cell proliferation, migration, invasion and metastasis, which is dependent on regulating the miR-548p/TUFT1 axis.

Recovery of phosphate and ammonia from wastewater via struvite precipitation using spent refractory brick gravel from steel industry.

Spent refractory brick (SRB) generated from the steel industry has a high magnesium content. In this study, a procedure was developed to utilize SRB gravels for efficient recovery of phosphate and ammonia from high strength wastewater via struvite (MgNH4PO4∙6H2O(s)) precipitation. Mg2+ and Ca2+ were first leached from SRB gravels using nitric acid solution. Ca2+ in the solution could inhibit struvite precipitation and was sequestered by dosing SO32- to form calcium sulfite (CaSO3(s)). The resulting Mg2+-rich solution was then employed to initiate struvite precipitation for phosphate and ammonia recovery. The optimal precipitation was achieved with a molar ratio of [Mg2+]:[NH3-N]:[PO43-P] = 2:1:2 at pH 9.5. The residual phosphate in the solution can be further removed via the precipitation of calcium phosphate minerals. Overall, 99.6% phosphate and 98.2% ammonia could be recovered and the treated wastewater could meet the discharging standards of ammonia and phosphate. The resulting solids, including calcium sulfite, struvite and calcium phosphate can be potentially used in the cement industry and agriculture sector to achieve sustainable recycle of spent materials.

Impact of Sequencing Depth and Library Preparation on Toxicological Interpretation of RNA-Seq Data in a "Three-Sample" Scenario.

While RNA-sequencing (RNA-seq) has emerged as a standard approach in toxicogenomics, its full potential in gaining underlying toxicological mechanisms is still not clear when only three biological replicates are used. This "three-sample" study design is common in toxicological research, particularly in animal studies during preclinical drug development. Sequencing depth (the total number of reads in an experiment) and library preparation are critical to the resolution and integrity of RNA-seq data and biological interpretation. We used aflatoxin b1 (AFB1), a model toxicant, to investigate the effect of sequencing depth and library preparation in RNA-seq on toxicological interpretation in the "three-sample" scenario. We also compared different gene profiling platforms (RNA-seq, TempO-seq, microarray, and qPCR) using identical liver samples. Well-established mechanisms of AFB1 toxicity served as ground truth for our comparative analyses. We found that a minimum of 20 million reads was sufficient to elicit key toxicity functions and pathways underlying AFB1-induced liver toxicity using three replicates and that identification of differentially expressed genes was positively associated with sequencing depth to a certain extent. Further, our results showed that RNA-seq revealed toxicological insights from pathway enrichment with overall higher statistical power and overlap ratio, compared with TempO-seq and microarray. Moreover, library preparation using the same methods was important to reproducing the toxicological interpretation.

Circular ribonucleic acid nei-like deoxyribonucleic acid glycosylase 3 governs the microribonucleic acid -3150b-3p/laminin subunit gamma 1 network to partially promote the development of hepatocellular carcinoma.

AIM: Hepatocellular carcinoma (HCC) is one of the most common malignant tumors, and its progression is implicated in the dysregulation of circular ribonucleic acids (RNAs). This study aimed to investigate the role of circular RNA nei-like DNA glycosylase 3 (circNEIL3) in HCC. METHODS: Real-time quantitative PCR was used for expression analysis of circNEIL3, microRNA-3150b-3p (miR-3150b-3p) and laminin subunit gamma 1 (LAMC1) message RNA. MTT assay, colony formation assay, wound healing assay, transwell assay, and flow cytometry assay were performed for functional analyses on cell proliferation, migration, invasion, apoptosis, and cycle. The expression of marker proteins and LAMC1 protein was quantified by western blot. The interaction between miR-3150b-3p and circNEIL3 or LAMC1 was confirmed by dual-luciferase reporter assay or RNA immunoprecipitation assay. An animal study was performed to confirm the role of circNEIL3 in vivo. RESULTS: CircNEIL3 was upregulated in tumor tissues and HCC cell lines. CircNEIL3 knockdown significantly suppressed HCC cell proliferation, migration and invasion and induced cell apoptosis and cell cycle arrest. MiR-3150b-3p was a target of circNEIL3, and its inhibition largely reversed the functional effects of circNEIL3 knockdown on cell behaviors. Moreover, LAMC1 served as a target of miR-3150b-3p, and its expression was elevated in HCC tissues and cells. LAMC1 overexpression recovered HCC cell proliferation, migration and invasion that were blocked by miR-3150b-3p restoration. Additionally, circNEIL3 knockdown inhibited tumor growth in mice. CONCLUSION: CircNEIL3 dysregulation was responsible for the partial development of HCC by regulating the miR-3150b-3p/LAMC1 regulatory network.

Coordinated Regulation of UGT2B15 Expression by Long Noncoding RNA LINC00574 and hsa-miR-129-5p in HepaRG Cells.

Recent studies have shown that microRNAs and long noncoding RNAs (lncRNAs) regulate the expression of drug metabolizing enzymes (DMEs) in human hepatic cells and that a set of DMEs, including UDP glucuronosyltransferase (UGT) 2B15, is down-regulated dramatically in liver cells by toxic acetaminophen (APAP) concentrations. In this study we analyzed mRNA, microRNA, and lncRNA expression profiles in APAP-treated HepaRG cells to explore noncoding RNA-dependent regulation of DME expression. The expression of UGT2B15 and lncRNA LINC00574 was decreased in APAP-treated HepaRG cells. UGT2B15 levels were diminished by LINC00574 suppression using antisense oligonucleotides or small interfering RNA. Furthermore, we found that hsa-miR-129-5p suppressed LINC00574 and decreased UGT2B15 expression via LINC00574 in HepaRG cells. In conclusion, our results indicate that LINC00574 acts as an important regulator of UGT2B15 expression in human hepatic cells, providing experimental evidence and new clues to understand the role of cross-talk between noncoding RNAs. SIGNIFICANCE STATEMENT: We showed a molecular network that displays the cross-talk and consequences among mRNA, micro RNA, long noncoding RNA, and proteins in acetaminophen (APAP)-treated HepaRG cells. APAP treatment increased the level of hsa-miR-129-5p and decreased that of LINC00574, ultimately decreasing the production of UDP glucuronosyltransferase (UGT) 2B15. The proposed regulatory network suppresses UGT2B15 expression through interaction of hsa-miR-129-5p and LINC00574, which may be achieved potentially by recruiting RNA binding proteins.

The role of hepatic cytochrome P450s in the cytotoxicity of sertraline.

Sertraline, an antidepressant, is commonly used to manage mental health symptoms related to depression, anxiety disorders, and obsessive-compulsive disorder. The use of sertraline has been associated with rare but severe hepatotoxicity. Previous research demonstrated that mitochondrial dysfunction, apoptosis, and endoplasmic reticulum stress were involved in sertraline-associated cytotoxicity. In this study, we reported that after a 24-h treatment in HepG2 cells, sertraline caused cytotoxicity, suppressed topoisomerase I and IIα, and damaged DNA in a concentration-dependent manner. We also investigated the role of cytochrome P450 (CYP)-mediated metabolism in sertraline-induced toxicity using our previously established HepG2 cell lines individually expressing 14 CYPs (1A1, 1A2, 1B1, 2A6, 2B6, 2C8, 2C9, 2C18, 2C19, 2D6, 2E1, 3A4, 3A5, and 3A7). We demonstrated that CYP2D6, 2C19, 2B6, and 2C9 metabolize sertraline, and sertraline-induced cytotoxicity was significantly decreased in the cells expressing these CYPs. Western blot analysis demonstrated that the induction of É£H2A.X (a hallmark of DNA damage) and topoisomerase inhibition were partially reversed in CYP2D6-, 2C19-, 2B6-, and 2C9-overexpressing HepG2 cells. These data indicate that DNA damage and topoisomerase inhibition are involved in sertraline-induced cytotoxicity and that CYPs-mediated metabolism plays a role in decreasing the toxicity of sertraline.

Long noncoding RNA LINC00844-mediated molecular network regulates expression of drug metabolizing enzymes and nuclear receptors in human liver cells.

Noncoding RNAs, such as long noncoding RNAs (lncRNAs) and microRNAs (miRNAs), regulate gene expression in many physiological and pathological processes, including drug metabolism. Drug metabolizing enzymes (DMEs) are critical components in drug-induced liver toxicity. In this study, we used human hepatic HepaRG cells treated with 5 or 10 mM acetaminophen (APAP) as a model system and identified LINC00844 as a toxicity-responsive lncRNA. We analyzed the expression profiles of LINC00844 in different human tissues. In addition, we examined the correlations between the levels of LINC00844 and those of key DMEs and nuclear receptors (NRs) for APAP metabolism in humans. Our results showed that lncRNA LINC00844 is enriched in the liver and its expression correlates positively with mRNA levels of CYP3A4, CYP2E1, SULT2A1, pregnane X receptor (PXR), and hepatocyte nuclear factor (HNF) 4α. We demonstrated that LINC00844 regulates the expression of these five genes in HepaRG cells using gain- and loss-of-function assays. Further, we discovered that LINC00844 is localized predominantly in the cytoplasm and acts as an hsa-miR-486-5p sponge, via direct binding, to protect SULT2A1 from miRNA-mediated gene silencing. Our data also demonstrated a functional interaction between LINC00844 and hsa-miR-486-5p in regulating DME and NR expression in HepaRG cells and primary human hepatocytes. We depicted a LINC00844-mediated regulatory network that involves miRNA and NRs and influences DME expression in response to APAP toxicity.

Membrane-associated androgen receptor (AR) potentiates its transcriptional activities by activating heat shock protein 27 (HSP27).

The androgen receptor (AR) is a ligand-activated nuclear receptor that plays a critical role in normal prostate physiology, as well as in the development and progression of prostate cancer. In addition to the classical paradigm in which AR exerts its biological effects in the nucleus by orchestrating the expression of the androgen-regulated transcriptome, there is considerable evidence supporting a rapid, nongenomic activity mediated by membrane-associated AR. Although the genomic action of AR has been studied in depth, the molecular events governing AR transport to the plasma membrane and the downstream AR signaling cascades remain poorly understood. In this study, we report that AR membrane transport is microtubule-dependent. Disruption of the function of kinesin 5B (KIF5B), but not of kinesin C3 (KIFC3), interfered with AR membrane association and signaling. Co-immunoprecipitation and pulldown assays revealed that AR physically interacts with KIF5B and that androgen enhances this interaction. Furthermore, we show that heat shock protein 27 (HSP27) is activated by membrane-associated AR and that HSP27 plays an important role in mediating AR-mediated membrane-to-nuclear signal transduction. Together, these results indicate that AR membrane translocation is mediated by the microtubule cytoskeleton and the motor protein KIF5B. By activating HSP27, membrane-associated AR potentiates the transcriptional activity of nuclear AR. We conclude that disruption of AR membrane translocation may represent a potential strategy for targeting AR signaling therapeutically in prostate cancer.

Regulation of cytochrome P450 expression by microRNAs and long noncoding RNAs: Epigenetic mechanisms in environmental toxicology and carcinogenesis.

Environmental exposures to hazardous chemicals are associated with a variety of human diseases and disorders, including cancers. Phase I metabolic activation and detoxification reactions catalyzed by cytochrome P450 enzymes (CYPs) affect the toxicities of many xenobiotic compounds. Proper regulation of CYP expression influences their biological effects. Noncoding RNAs (ncRNAs) are involved in regulating CYP expression, and ncRNA expression is regulated in response to environmental chemicals. The mechanistic interactions between ncRNAs and CYPs associated with the toxicity and carcinogenicity of environmental chemicals are described in this review, focusing on microRNA-dependent CYP regulation. The role of long noncoding RNAs in regulating CYP expression is also presented and new avenues of research concerning this regulatory mechanism are described.