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Se Pu = Chinese Journal of... Oct 2022A new method for sample pretreatment using improved QuEChERs was established, and 289 organic pollutants with health risks could be identified and quantified through gas...
A new method for sample pretreatment using improved QuEChERs was established, and 289 organic pollutants with health risks could be identified and quantified through gas chromatography-orbitrap high-resolution mass spectrometry (GC-Orbitrap HRMS). A high-resolution database of 289 environmental pollutants belonging to ten categories, including organochlorine pesticides (OCPs), polycyclic aromatic hydrocarbons (PAHs), phthalates (PAEs), polychlorinated biphenyls (PCBs), and other agricultural chemicals (ACs), was established for the non-targeted screening and quantitative analysis. A simple method for biological sample preparation using improved QuEChERs was proposed by combining a conventional QuEChERs method and a column purification method. After purification using a Florisil column, the lipid content was reduced by 99.9%, which significantly reduced the interference of the matrix effect observed during the analysis. Furthermore, simultaneous high-accuracy qualitative screening and quantitative analysis of the target compounds were performed through high-resolution mass spectrometry (60000 resolution) conducted in the full scan mode. The limits of quantification were 0.56-57.8 pg/g, presenting a large linear range (~10), and the recovery range was 40%-120%. Due to the high-resolution and sensitivity of Q Exactive GC-Orbitrap HRMS, the limits of quantification of the target compounds were significantly lower than those achieved through methods based on conventional chromatography and mass spectrometry. Moreover, ultratrace organic contaminants that cannot be detected by conventional methods can be accurately quantified by the proposed method. Sea cucumber samples collected at the breeding site were analyzed using the proposed high-coverage multi-objective analytical method, and more than 100 types of organic pollutants were detected; the mean contents of PAHs, ACs, PAEs, and OCPs were 157.8, 153.2, 64.4, and 46.4 ng/g dw, respectively, which were higher than those of other pollutants. Some new contaminants, such as 9-chlorofluorene, 5-chloroacenaphthene, and 3-methylcholanthrene, were detected at very low contents for the first time in the sea cucumber samples. The proposed method is simple and efficient, allows the detection of pollutants at very low contents, and provides accurate and reliable results. Thus, this high-coverage multi-objective analytical method can be widely used for broad-spectrum screening and accurate quantification of contaminants in various aquatic products, providing technical support for food safety control.
Topics: Animals; Environmental Pollutants; Gas Chromatography-Mass Spectrometry; Hydrocarbons, Chlorinated; Lipids; Mass Spectrometry; Methylcholanthrene; Pesticides; Polychlorinated Biphenyls; Polycyclic Aromatic Hydrocarbons; Sea Cucumbers
PubMed: 36222258
DOI: 10.3724/SP.J.1123.2022.04001 -
Molecular Cancer Therapeutics Jan 2023This study aims to investigate whether adding neoadjuvant radiotherapy (RT), anti-programmed cell death protein-1 (PD-1) antibody (anti-PD-1), or RT + anti-PD-1 to...
This study aims to investigate whether adding neoadjuvant radiotherapy (RT), anti-programmed cell death protein-1 (PD-1) antibody (anti-PD-1), or RT + anti-PD-1 to surgical resection improves disease-free survival for mice with soft tissue sarcomas (STS). We generated a high mutational load primary mouse model of STS by intramuscular injection of adenovirus expressing Cas9 and guide RNA targeting Trp53 and intramuscular injection of 3-methylcholanthrene (MCA) into the gastrocnemius muscle of wild-type mice (p53/MCA model). We randomized tumor-bearing mice to receive isotype control or anti-PD-1 antibody with or without radiotherapy (20 Gy), followed by hind limb amputation. We used micro-CT to detect lung metastases with high spatial resolution, which was confirmed by histology. We investigated whether sarcoma metastasis was regulated by immunosurveillance by lymphocytes or tumor cell-intrinsic mechanisms. Compared with surgery with isotype control antibody, the combination of anti-PD-1, radiotherapy, and surgery improved local recurrence-free survival (P = 0.035) and disease-free survival (P = 0.005), but not metastasis-free survival. Mice treated with radiotherapy, but not anti-PD-1, showed significantly improved local recurrence-free survival and metastasis-free survival over surgery alone (P = 0.043 and P = 0.007, respectively). The overall metastasis rate was low (∼12%) in the p53/MCA sarcoma model, which limited the power to detect further improvement in metastasis-free survival with addition of anti-PD-1 therapy. Tail vein injections of sarcoma cells into immunocompetent mice suggested that impaired metastasis was due to inability of sarcoma cells to grow in the lungs rather than a consequence of immunosurveillance. In conclusion, neoadjuvant radiotherapy improves metastasis-free survival after surgery in a primary model of STS.
Topics: Mice; Animals; Neoadjuvant Therapy; Tumor Suppressor Protein p53; Sarcoma; Progression-Free Survival; Disease-Free Survival; Soft Tissue Neoplasms; Retrospective Studies; Radiotherapy, Adjuvant; Neoplasm Recurrence, Local
PubMed: 36162051
DOI: 10.1158/1535-7163.MCT-21-0991 -
Journal of Immunology Research 2022Tumor immune evasion is a lineament of cancer. Endothelial monocyte activating polypeptide-II (EMAP II) has been assumed to impact tumor immune escape significantly....
Tumor immune evasion is a lineament of cancer. Endothelial monocyte activating polypeptide-II (EMAP II) has been assumed to impact tumor immune escape significantly. EMAP II was first reported in the murine methylcholanthrene A-induced fibrosarcoma supernatant and identified as a tumor-derived cytokine. This study evaluated EMAP II expression in peripheral blood cells and its association with treatment outcome, lactate dehydrogenase (LDH) levels, and clinical criteria in non-Hodgkin's lymphoma (NHL) patients. EMAP II expression on different blood cells obtained from the peripheral blood of 80 NHL patients was evaluated by two-color flow cytometry. The study reported that EMAP II expression was significantly increased in peripheral blood cells in patients with NHL compared to normal volunteers ( < 0.001). Additionally, EMAP II expression levels on blood cells decreased in complete remission (CR) while they increased in relapse. This study showed coexpression of EMAP II and CD36 on peripheral lymphocytes in NHL patients but not in healthy controls ( < 0.001). EMAP II expression on blood cells was associated with increased serum LDH levels. Furthermore, the percentages of EMAP II+/CD36+ peripheral lymphocytes were significantly higher in relapse than in CR and healthy controls. Analyses revealed that higher percentages of EMAP II+CD36+ cells were positively correlated with hepatomegaly, splenomegaly, and an advanced (intermediate and high risk) NHL stage. The results assume that EMAP II might be involved in NHL development and pathogenesis.
Topics: Animals; Cytokines; Humans; Lactate Dehydrogenases; Lymphocytes; Lymphoma, Non-Hodgkin; Methylcholanthrene; Mice; Neoplasm Proteins; Neoplasm Recurrence, Local; RNA-Binding Proteins
PubMed: 36132984
DOI: 10.1155/2022/7219207 -
Biological & Pharmaceutical Bulletin 2022The aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor that mediates the toxicity of dioxins and polycyclic aromatic hydrocarbons. Recent studies...
The aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor that mediates the toxicity of dioxins and polycyclic aromatic hydrocarbons. Recent studies have suggested that AhR is involved in cancer immunity. In the present study, we examined whether AhR regulates the expression of immune checkpoint genes in breast cancer cells. We discovered that the mRNA expression of V-set domain containing T cell activation inhibitor 1 (VTCN1) that negatively regulates T cell immunity was upregulated by AhR agonists in breast cancer cell lines, MCF-7 and T47D. Furthermore, AhR knockout or knockdown experiments clearly demonstrated that upregulation of VTCN1 gene expression by 3-methylcholanthrene was AhR dependent. Luciferase reporter and chromatin immunoprecipitation assays revealed that this upregulation of VTCN1 gene expression was induced by the recruitment of AhR to the AhR responsive element in the VTCN1 gene promoter in MCF-7 cells. Taken together, AhR directly regulates VTCN1 gene expression in MCF-7 cells.
Topics: Basic Helix-Loop-Helix Transcription Factors; Breast Neoplasms; Female; Gene Expression; Humans; MCF-7 Cells; Methylcholanthrene; Receptors, Aryl Hydrocarbon; V-Set Domain-Containing T-Cell Activation Inhibitor 1
PubMed: 35650105
DOI: 10.1248/bpb.b21-01068 -
International Journal of Medical... 2022There is growing support for the notion that chronic inflammation contributes to lung tumorigenesis, but the molecular and cellular basis underlying the protumorigenic...
There is growing support for the notion that chronic inflammation contributes to lung tumorigenesis, but the molecular and cellular basis underlying the protumorigenic effects of inflammation remains to be explored. 3-Methylcholanthrene and diethylnitrosamine were intratracheally instilled into rats to induce multistep lung carcinogenesis, and the presence of pulmonary inflammation was observed in addition to precancerous lesions. By leveraging single-cell RNA sequencing, we sought to unravel the mechanism underlying the inflammatory process at a higher resolution. A total of 14 cell types were identified in chemically treated and control rats. Chemical intervention introduced heterogeneity in cell type composition and gene expression patterns. Nonimmune cells were found to be the most affected, and two subpopulations of endothelial cells with diverse roles were defined. Car4-high endothelial cells were mainly responsible for angiogenesis, whereas Car4-low endothelial cells were involved in neutrophil recruitment, and adhesion between Car4-low endothelial cells and neutrophils was verified in inflamed tissues. Our work unveiled the intricate process of pulmonary inflammation at the single-cell level and characterized a proinflammatory subpopulation of endothelial cells involved in neutrophil recruitment. The conditions provided by chronic inflammatory environment are prerequisites for neoplastic progression. Targeting the specific subsets or processes defined herein holds promise for the early prevention and therapeutic intervention of lung cancer through the manipulation of angiogenesis or the inflammatory response.
Topics: Animals; Endothelial Cells; Inflammation; Neutrophil Infiltration; Neutrophils; Pneumonia; Rats; Sequence Analysis, RNA
PubMed: 35582423
DOI: 10.7150/ijms.67806 -
Frontiers in Pharmacology 2022Acquired cisplatin resistance in cervical cancer therapy is principally caused by reduction in intracellular drug accumulation, which is exerted by hyperactivation of...
Acquired cisplatin resistance in cervical cancer therapy is principally caused by reduction in intracellular drug accumulation, which is exerted by hyperactivation of the oncogenic PI3K/Akt signaling axis and overexpression of cisplatin-exporter MRP2 along with prosurvival effectors NF-κB and IAPs in cervical cancer cells. These activated prosurvival signaling cascades drive drug efflux and evasion of apoptosis for rendering drug-resistant phenotypes. Our study challenges the PI3K/Akt axis in a cisplatin-resistant cervical cancer scenario with phenethylisothiocyanate (PEITC) for chemosensitization of SiHa, a cisplatin-resistant sub-line of SiHa and 3-methylcholanthrene-induced cervical cancer mice models. SiHa exhibited higher MRP2, p-Akt, NF-κB, XIAP, and survivin expressions which cumulatively compromised cisplatin retention capacity and accumulated PEITC better than SiHa. SiHa appeared to favor PEITC uptake as its accumulation rates were found to be positively correlated with MRP2 expressions. PEITC treatment in SiHa for 3 h prior to cisplatin exposure revived intracellular platinum levels, reduced free GSH levels, generated greater ROS, and altered mitochondrial membrane potential compared to SiHa. Western blot and immunofluorescence results indicated that PEITC successfully downregulated MRP2 in addition to suppressing p-Akt, XIAP, survivin, and NF-κB expressions. In mice models, administration of 5 mg/kg body-weight PEITC priming dosage prior to treatment with 3 mg/kg body-weight of cisplatin remediated cervical histology and induced tumor regression in contrast to the group receiving the same dosage of cisplatin only. This suggested PEITC as a potential chemosensitizing agent in light of acquired cisplatin resistance in cervical cancer and established its candidature for Phase I clinical trial.
PubMed: 35548339
DOI: 10.3389/fphar.2022.803114 -
Current Research in Toxicology 2022Polycyclic aromatic compounds (PACs) are a broad class of contaminants ubiquitously present in the environment due to natural and anthropogenic activities. With...
Polycyclic aromatic compounds (PACs) are a broad class of contaminants ubiquitously present in the environment due to natural and anthropogenic activities. With increasing industrialization and reliance on petroleum worldwide, PACs are increasingly being detected in different environmental compartments. Previous studies have shown that PACs possess endocrine disruptive properties as these compounds often interfere with hormone signaling and function. In females, the ovary is largely responsible for regulating reproductive and endocrine function and thus, serves as a primary target for PAC-mediated toxicity. Perturbations in the signaling pathways that mediate ovarian folliculogenesis, steroidogenesis and angiogenesis can lead to adverse reproductive outcomes including polycystic ovary syndrome, premature ovarian insufficiency, and infertility. To date, the impact of PACs on ovarian function has focused predominantly on polycyclic aromatic hydrocarbons like benzo(a)pyrene, 3-methylcholanthrene and 7,12-dimethylbenz[a]anthracene. However, investigation into the impact of substituted PACs including halogenated, heterocyclic, and alkylated PACs on mammalian reproduction has been largely overlooked despite the fact that these compounds are found in higher abundance in free-ranging wildlife. This review aims to discuss current literature on the effects of PACs on the ovary in mammals, with a particular focus on folliculogenesis, steroidogenesis and angiogenesis, which are key processes necessary for proper ovarian functions.
PubMed: 35492299
DOI: 10.1016/j.crtox.2022.100070 -
International Journal of Molecular... Dec 2021Cutaneous squamous cell carcinoma (cSCC) accounts for 25% of cutaneous malignancies diagnosed in Caucasian populations. Surgical removal in combination with radiation...
Cutaneous squamous cell carcinoma (cSCC) accounts for 25% of cutaneous malignancies diagnosed in Caucasian populations. Surgical removal in combination with radiation and chemotherapy are effective treatments for cSCC. Nevertheless, the aggressive metastatic forms of cSCC still have a relatively poor patient outcome. Studies have linked actin cytoskeletal dynamics and the Wnt/β-catenin signaling pathway as important modulators of cSCC pathogenesis. Previous studies have also shown that the actin-remodeling protein Flightless (Flii) is a negative regulator of cSCC. The aim of this study was to investigate if the functional effects of Flii on cSCC involve the Wnt/β-catenin signaling pathway. Flii knockdown was performed using siRNA in a human late stage aggressive metastatic cSCC cell line (MET-1) alongside analysis of genetic murine models of 3-methylcholanthrene induced cSCC. Flii was increased in a MET-1 cSCC cell line and reducing Flii expression led to fewer PCNA positive cells and a concomitant reduction in cellular proliferation and symmetrical division. Knockdown of Flii led to decreased β-catenin and a decrease in the expression of the downstream effector of β-catenin signaling protein SOX9. 3-Methylcholanthrene (MCA)-induced cSCC in overexpressing mice showed increased markers of cancer metastasis including talin and keratin-14 and a significant increase in SOX9 alongside a reduction in Flii associated protein (Flap-1). Taken together, this study demonstrates a role for Flii in regulating proteins involved in cSCC proliferation and tumor progression and suggests a potential role for Flii in aggressive metastatic cSCC.
Topics: Animals; Carcinoma, Squamous Cell; Cell Line, Tumor; Cell Proliferation; Disease Models, Animal; Female; Gene Expression Regulation, Neoplastic; Gene Knockdown Techniques; Humans; Methylcholanthrene; Mice; Microfilament Proteins; Skin Neoplasms; Trans-Activators; Up-Regulation; Wnt Signaling Pathway
PubMed: 34948000
DOI: 10.3390/ijms222413203 -
Frontiers in Cellular and Infection... 2021The opportunistic pathogen is one of the few intestinal bacteria that has been consistently linked to colorectal cancer (CRC). This study aimed to identify novel...
OBJECTIVE
The opportunistic pathogen is one of the few intestinal bacteria that has been consistently linked to colorectal cancer (CRC). This study aimed to identify novel -induced pathways in colon epithelial cells that could further explain how contributes to CRC development.
DESIGN AND RESULTS
Transcription profiling of cultured CRC cells that were exposed to revealed the specific induction of oxidoreductase pathways. Most prominently, and genes that encode phase I biotransformation enzymes were responsible for the detoxification or bio-activation of toxic compounds. A common feature is that these enzymes are induced through the Aryl hydrocarbon receptor (AhR). Using the specific inhibitor CH223191, we showed that the induction of was dependent on the AhR both using multiple CRC cell lines as using wild-type C57bl6 mice colonized with . Furthermore, we showed that CYP1 could also be induced by other intestinal bacteria and that a yet unidentified diffusible factor from the secretome (SGS) induces CYP1A enzyme activity in an AhR-dependent manner. Importantly, priming CRC cells with SGS increased the DNA damaging effect of the polycyclic aromatic hydrocarbon 3-methylcholanthrene.
CONCLUSION
This study shows that gut bacteria have the potential to modulate the expression of biotransformation pathways in colonic epithelial cells in an AhR-dependent manner. This offers a novel theory on the contribution of intestinal bacteria to the etiology of CRC by modifying the capacity of intestinal epithelial or (pre-)cancerous cells to (de)toxify dietary components, which could alter intestinal susceptibility to DNA damaging events.
Topics: Animals; Biotransformation; Colorectal Neoplasms; Cytochrome P-450 CYP1A1; Epithelial Cells; Mice; Mice, Inbred C57BL; Receptors, Aryl Hydrocarbon; Streptococcus gallolyticus
PubMed: 34778104
DOI: 10.3389/fcimb.2021.740704 -
Archives of Toxicology Jan 2022Error-corrected sequences (ECSs) that utilize double-stranded DNA sequences are useful in detecting mutagen-induced mutations. However, relatively higher frequencies of...
Error-corrected sequences (ECSs) that utilize double-stranded DNA sequences are useful in detecting mutagen-induced mutations. However, relatively higher frequencies of G:C > T:A (1 × 10 bp) and G:C > C:G (2 × 10 bp) errors decrease the accuracy of detection of rare G:C mutations (approximately 10 bp). Oxidized guanines in single-strand (SS) overhangs generated after shearing could serve as the source of these errors. To remove these errors, we first computationally discarded up to 20 read bases corresponding to the ends of the DNA fragments. Error frequencies decreased proportionately with trimming length; however, the results indicated that they were not sufficiently removed. To efficiently remove SS overhangs, we evaluated three mechanistically distinct SS-specific nucleases (S1 Nuclease, mung bean nuclease, and RecJf exonuclease) and found that they were more efficient than computational trimming. Consequently, we established Jade-Seq™, an ECS protocol with S1 Nuclease treatment, which reduced G:C > T:A and G:C > C:G errors to 0.50 × 10 bp and 0.12 × 10 bp, respectively. This was probably because S1 Nuclease removed SS regions, such as gaps and nicks, depending on its wide substrate specificity. Subsequently, we evaluated the mutation-detection sensitivity of Jade-Seq™ using DNA samples from TA100 cells exposed to 3-methylcholanthrene and 7,12-dimethylbenz[a]anthracene, which contained the rare G:C > T:A mutation (i.e., 2 × 10 bp). Fold changes of G:C > T:A compared to the vehicle control were 1.2- and 1.3-times higher than those of samples without S1 Nuclease treatment, respectively. These findings indicate the potential of Jade-Seq™ for detecting rare mutations and determining the mutagenicity of environmental mutagens.
Topics: DNA; DNA Repair; High-Throughput Nucleotide Sequencing; Mutagens; Mutation
PubMed: 34767040
DOI: 10.1007/s00204-021-03185-y