Trajectory of medical expenditure and regional disparities in hypertensive patients in South Korea.

The aim of this study is to understand how different regions influence the management and financial burden of hypertension, and to identify regional disparities in hypertension management and medical expenditure. The study utilized data from the Korean Health Panel Survey conducted between 2014 and 2018, focusing on individuals with hypertension. Medical expenditures were classified into three trajectory groups: "Persistent Low," "Expenditure Increasing," and "Persistent High" over a five-year period using trajectory analysis. Inverse Probability Weighting (IPW) analysis was then employed to identify the association between regions and medical expenditure trajectories. The results indicate that individuals residing in metropolitan cities (Busan, Daegu, Incheon, Gwangju, Daejeon, and Ulsan) and rural areas were more likely to belong to the "Expenditure Increasing" group compared to the "Persistent Low Expenditure" group (OR = 1.07; 95% CI; p < 0.001), as opposed to those in the capital city (Seoul) (OR = 1.07; 95% CI; p < 0.001). Additionally, residents of rural areas were more likely to be in the "High Expenditure" group compared to the "Persistent Low Expenditure" group than those residing in the capital city (OR = 1.05; 95% CI; p = 0.001). These findings suggest that individuals in rural areas may be receiving relatively inadequate management for hypertension, leading to higher medical expenditures compared to those in the capital region. These disparities signify health inequality and highlight the need for policy efforts to address regional imbalances in social structures and healthcare resource distribution to ensure equitable chronic disease management across different regions.

Osthole Suppresses Cell Growth of Prostate Cancer by Disrupting Redox Homeostasis, Mitochondrial Function, and Regulation of tiRNAHisGTG.

Prostate cancer remains a significant global health concern, posing a substantial threat to men's well-being. Despite advancements in treatment modalities, the progression of prostate cancer still presents challenges, warranting further exploration of novel therapeutic strategies. In this study, osthole, a natural coumarin derivative, inhibited cell viability in cancer cells but not in the normal prostate cell line. Moreover, osthole disrupted cell cycle progression. Furthermore, osthole reduces mitochondrial respiration with mitochondrial membrane potential (ΔΨm) depolarization and reactive oxygen species (ROS) generation, indicating mitochondrial dysfunction. In particular, osthole-induced ROS generation was reduced by N-acetyl-L-cysteine (NAC) in prostate cancer. In addition, using calcium inhibitors (2-APB and ruthenium red) and endoplasmic reticulum (ER) stress inhibitor (4-PBA), we confirmed that ER stress-induced calcium overload by osthole causes mitochondrial dysfunction. Moreover, we verified that the osthole-induced upregulation of tiRNAHisGTG expression is related to mechanisms that induce permeabilization of the mitochondrial membrane and calcium accumulation. Regarding intracellular signaling, osthole inactivated the PI3K and ERK pathways while activating the expression of the P38, JNK, ER stress, and autophagy-related proteins. In conclusion, the results suggest that osthole can be used as a therapeutic or adjuvant treatment for the management of prostate cancer.

Image segmentation of impacted mesiodens using deep learning.

This study aimed to evaluate the performance of deep learning algorithms for the classification and segmentation of impacted mesiodens in pediatric panoramic radiographs. A total of 850 panoramic radiographs of pediatric patients (aged 3-9 years) was included in this study. The U-Net semantic segmentation algorithm was applied for the detection and segmentation of mesiodens in the upper anterior region. For enhancement of the algorithm, pre-trained ResNet models were applied to the encoding path. The segmentation performance of the algorithm was tested using the Jaccard index and Dice coefficient. The diagnostic accuracy, precision, recall, F1-score and time to diagnosis of the algorithms were compared with those of human expert groups using the test dataset. Cohen's kappa statistics were compared between the model and human groups. The segmentation model exhibited a high Jaccard index and Dice coefficient (>90%). In mesiodens diagnosis, the trained model achieved 91-92% accuracy and a 94-95% F1-score, which were comparable with human expert group results (96%). The diagnostic duration of the deep learning model was 7.5 seconds, which was significantly faster in mesiodens detection compared to human groups. The agreement between the deep learning model and human experts is moderate (Cohen's kappa = 0.767). The proposed deep learning algorithm showed good segmentation performance and approached the performance of human experts in the diagnosis of mesiodens, with a significantly faster diagnosis time.

Color stability and translucency of newly developed composites for splinting teeth.

This study aimed to investigate the colorimetric properties of newly developed composites for dental trauma splints using various staining solutions during the clinical splinting period. The clear shades of G-Fix (GF), Ortho Connect Flow (OC), Light Fix (LF), and Filtek Z350XT (FZ) were fabricated into 96 disk-shaped specimens. Specimens from each composite group were stored in distilled water, coffee, tea, and red wine solutions at 37ºC. CIE values were measured using a spectrophotometer at 24 h after specimen preparation and at 1 day, 1 week, 2 weeks, 3 weeks, and 4 weeks after storage in each solution. Color differences and translucency parameters were calculated using the initial and measured values. Within the experiment period, the color differences of GF, OC, and LF compared to the initial measurement were smaller than that for FZ for all staining solutions except distilled water. There were no significant color differences between the GF, OC, and LF groups.

Fully automated deep learning approach to dental development assessment in panoramic radiographs.

BACKGROUND: Dental development assessment is an important factor in dental age estimation and dental maturity evaluation. This study aimed to develop and evaluate the performance of an automated dental development staging system based on Demirjian's method using deep learning. METHODS: The study included 5133 anonymous panoramic radiographs obtained from the Department of Pediatric Dentistry database at Seoul National University Dental Hospital between 2020 and 2021. The proposed methodology involves a three-step procedure for dental staging: detection, segmentation, and classification. The panoramic data were randomly divided into training and validating sets (8:2), and YOLOv5, U-Net, and EfficientNet were trained and employed for each stage. The models' performance, along with the Grad-CAM analysis of EfficientNet, was evaluated. RESULTS: The mean average precision (mAP) was 0.995 for detection, and the segmentation achieved an accuracy of 0.978. The classification performance showed F1 scores of 69.23, 80.67, 84.97, and 90.81 for the Incisor, Canine, Premolar, and Molar models, respectively. In the Grad-CAM analysis, the classification model focused on the apical portion of the developing tooth, a crucial feature for staging according to Demirjian's method. CONCLUSIONS: These results indicate that the proposed deep learning approach for automated dental staging can serve as a supportive tool for dentists, facilitating rapid and objective dental age estimation and dental maturity evaluation.

Correlation between dental and skeletal maturity in Korean children based on dental maturity percentile: a retrospective study.

BACKGROUND: The correlation between dental maturity and skeletal maturity has been proposed, but its clinical application remains challenging. Moreover, the varying correlations observed in different studies indicate the necessity for research tailored to specific populations. AIM: To compare skeletal maturity in Korean children with advanced and delayed dental maturity using dental maturity percentile. DESIGN: Dental panoramic radiographs and cephalometric radiographs were obtained from 5133 and 395 healthy Korean children aged between 4 and 16 years old. Dental maturity was assessed with Demirjian's method, while skeletal maturity was assessed with the cervical vertebral maturation method. Standard percentile curves were developed through quantile regression. Advanced (93 boys and 110 girls) and delayed (92 boys and 100 girls) dental maturity groups were defined by the 50th percentile. RESULTS: The advanced group showed earlier skeletal maturity in multiple cervical stages (CS) in both boys (CS 1, 2, 3, 4, and 6) and girls (CS 1, 3, 4, 5, and 6). Significant differences, as determined by Mann-Whitney U tests, were observed in CS 1 for boys (p = 0.004) and in CS 4 for girls (p = 0.037). High Spearman correlation coefficients between dental maturity and cervical vertebral maturity exceeded 0.826 (p = 0.000) in all groups. CONCLUSION: A correlation between dental and skeletal maturity, as well as advanced skeletal maturity in the advanced dental maturity group, was observed. Using percentile curves to determine dental maturity may aid in assessing skeletal maturity, with potential applications in orthodontic diagnosis and treatment planning.

Autophagy regulation and redox perturbation by transcrocetin suppress the growth of endometriosis.

Until non-hormonal therapeutic targets for endometriosis are suggested, we focused on mitochondrial function and autophagy regulation in the disease. Transcrocetin is a carotenoid and retinoic acid with high antioxidant potency and antiproliferative effects in several diseases. In this study, we demonstrated the therapeutic mechanisms of transcrocetin in endometriosis using the End1/E6E7 and VK2/E6E7 cell lines. Transcrocetin suppressed the viability and proliferation of these cell lines and did not affect the proliferation of normal uterine stromal cells. p21 Waf1/Cip1 as a cell cycle regulator and target of p53, were increased by transcrocetin and caused the G1 arrest via inhibition of cyclin-dependent kinase activity, which might further cause cell death. Furthermore, we confirmed endoplasmic reticulum stress and calcium ion dysregulation in the cytosol and mitochondrial matrix, disrupting the mitochondrial membrane potential. Mitochondrial bioenergetics were suppressed by transcrocetin, and oxidative phosphorylation-related gene expression was downregulated. Moreover, the proliferation of End1/E6E7 and VK2/E6E7 cells was regulated by transcrocetin-induced oxidative stress. Finally, we verified the impairment of autophagic flux following pre-treatment with chloroquine. Therefore, transcrocetin may be a potent therapeutic alternative for endometriosis.

Relevance of the endoplasmic reticulum-mitochondria axis in cancer diagnosis and therapy.

Dynamic interactions between organelles are responsible for a variety of intercellular functions, and the endoplasmic reticulum (ER)-mitochondrial axis is recognized as a representative interorganelle system. Several studies have confirmed that most proteins in the physically tethered sites between the ER and mitochondria, called mitochondria-associated ER membranes (MAMs), are vital for intracellular physiology. MAM proteins are involved in the regulation of calcium homeostasis, lipid metabolism, and mitochondrial dynamics and are associated with processes related to intracellular stress conditions, such as oxidative stress and unfolded protein responses. Accumulating evidence has shown that, owing to their extensive involvement in cellular homeostasis, alterations in the ER-mitochondrial axis are one of the etiological factors of tumors. An in-depth understanding of MAM proteins and their impact on cell physiology, particularly in cancers, may help elucidate their potential as diagnostic and therapeutic targets for cancers. For example, the modulation of MAM proteins is utilized not only to target diverse intracellular signaling pathways within cancer cells but also to increase the sensitivity of cancer cells to anticancer reagents and regulate immune cell activities. Therefore, the current review summarizes and discusses recent advances in research on the functional roles of MAM proteins and their characteristics in cancers from a diagnostic perspective. Additionally, this review provides insights into diverse therapeutic strategies that target MAM proteins in various cancer types.

Osthole impairs mitochondrial metabolism and the autophagic flux in colorectal cancer.

BACKGROUND: Osthole is active constituent of Cnidium monnieri (L.) Cuss. with various physiological functions including anti-inflammation and anti-lipedemic effects. However, the regulatory activity of osthole in colorectal cancer development, focusing on mitochondrial metabolism, is not well known. HYPOTHESIS/PURPOSE: We hypothesized that osthole may suppress progression of colorectal cancer and aimed to determine the underlying mitochondrial metabolism and the autophagic flux. STUDY DESIGN: In this study, we elucidated the mechanism of action of osthole in colorectal cancer using an in vivo azoxymethane/dextran sodium sulfate (AOM/DSS) mouse model and an in vitro cell culture system. METHODS: AOM/DSS mouse model was established and analyzed the effects of osthole on survival rate, diseases activity index, number of tumor and histopathology. Then, cell based assays including viability, cell cycle, reactive oxygen species (ROS), apoptosis, calcium efflux, and mitochondrial function were analyzed. Moreover, osthole-mediated signaling was demonstrated by western blot analyses. RESULTS: Osthole effectively suppressed the growth of colorectal tumors and alleviated AOM/DSS-induced intestinal injury. Osthole restored the function of goblet cells and impaired the expression of Claudin1 and Axin1 impaired by AOM/DSS. In addition, osthole specifically showed cytotoxicity in colorectal carcinoma cells, but not in normal colon cells. Osthole decreased the ASC/caspase-1/IL-1ß inflammasome pathway and induced mitochondrial dysfunction in redox homeostasis, calcium homeostasis. Furthermore, osthole inhibited both oxidative phosphorylation (OXPHOS) and glycolysis, leading to the suppression of ATP production. Moreover, via combination treatment with chloroquine (CQ), we demonstrated that osthole impaired autophagic flux, leading to apoptosis of HCT116 and HT29 cells. Finally, we elucidated that the functional role of tiRNAHisGTG regulated by osthole directly affects the cellular fate of colon cancer cells. CONCLUSION: These results suggest that osthole has the potential to manage progression of colorectal cancer by regulating autophagy- and mitochondria-mediated signal transduction.

Fraxetin reduces endometriotic lesions through activation of ER stress, induction of mitochondria-mediated apoptosis, and generation of ROS.

BACKGROUND: Fraxetin, a phytochemical obtained from Fraxinus rhynchophylla, is well known for its anti-inflammatory and anti-fibrotic properties. However, fraxetin regulates the progression of endometriosis, which is a benign reproductive disease that results in low quality of life and infertility. HYPOTHESIS/PURPOSE: We hypothesized that fraxetin may have therapeutic effects on endometriosis and aimed to elucidate the underlying mechanisms of mitochondrial function and tiRNA regulation. STUDY DESIGN: Endometriotic animal models and cells (End1/E6E7 and VK2/E6E7) were used to identify the mode of action of fraxetin. METHODS: An auto-implanted endometriosis animal model was established and the effects of fraxetin on lesion size reduction were analyzed. Cell-based assays including proliferation, cell cycle, migration, apoptosis, mitochondrial function, calcium efflux, and reactive oxygen species (ROS) were performed. Moreover, fraxetin signal transduction was demonstrated by western blotting and qPCR analyses. RESULTS: Fraxetin inhibited proliferation and migration by inactivating the P38/JNK/ERK mitogen-activated protein kinase (MAPK) and AKT/S6 pathways. Fraxetin dissipates mitochondrial membrane potential, downregulates oxidative phosphorylation (OXPHOS), and disrupts redox and calcium homeostasis. Moreover, it triggered endoplasmic reticulum stress and intrinsic apoptosis. Furthermore, we elucidated the functional role of tiRNAHisGTG in endometriosis by transfection with its inhibitor. Finally, we established an endometriosis mouse model and verified endometriotic lesion regression and downregulation of adhesion molecules with inflammation. CONCLUSION: This study suggests that fraxetin is a novel therapeutic agent that targets mitochondria and tiRNAs. This is the first study to demonstrate the mechanisms of tiRNAHisGTG with mitochondrial function and cell fates and can be applied as a non-hormonal method against the progression of endometriosis.

Criteria for early diagnosis of mandibular third molar agenesis based on the developmental stages of mandibular canine, first and second premolars, and second molar: a retrospective cohort study.

BACKGROUND: Permanent first molars with severe dental caries, developmental defects, or involved in oral pathologies are at risk of poor prognosis in children. Accordingly, using the third molar to replace the first molar can be a good treatment option when third molar agenesis is predicted early. Thus, this retrospective cohort study aimed to develop criteria for early detection of mandibular third molar (L8) agenesis based on the developmental stages of mandibular canine (L3), first premolar (L4), second premolar (L5), and second molar (L7). METHOD: Overall, 1,044 and 919 panoramic radiographs of 343 males and 317 females, respectively, taken between the ages of 6 and 12 years were included. All developmental stages of L3, L4, L5, L7, and L8 were analyzed based on the dental age, as suggested by Demirjian et al. The independent t-test was used to assess age differences between males and females. The rank correlation coefficients were examined using Kendall's tau with bootstrap analysis and Bonferroni's correction to confirm the teeth showing developmental stages most similar to those of L8s. Finally, a survival analysis was performed to determine the criteria for the early diagnosis of mandibular third molar agenesis. RESULTS: Some age differences were found in dental developmental stages between males and females. Correlation coefficients between all stages of L3, L4, L5, and L7 and L8 were high. In particular, the correlation coefficient between L7 and L8 was the highest, whereas that between L3 and L8 was the lowest. CONCLUSION: If at least two of the following criteria (F stage of L3, F stage of L4, F stage of L5, and E stage of L7) are met in the absence of L8 crypt, agenesis of L8 can be confirmed.

The pattern of intra-/inter-hemispheric interactions of left and right hemispheres in visual word processing.

This study aimed to investigate the intra-/inter-hemispheric interactions during visual word processing, by manipulating stimulus onset asynchrony (SOA) in a primed-lateralized lexical decision task. To assess intra-/inter-hemispheric priming effects, identical prime-target pairs were presented in the same or opposite unilateral visual fields. The study found that the right visual field advantage (RVFA) was observed when Korean words were presented sequentially within hemispheres, indicating that the inherent characteristics of the two hemispheres, rather than differences in memory or linguistic aspects of lexical processing, contributed to the hemispheric asymmetry. Additionally, intra-hemispheric priming effects were symmetrical in both hemispheres, with similar increases in priming for words and nonwords from SOA 120 ms to SOA 600 ms. Furthermore, inter-hemispheric priming effects were asymmetrical, with stronger priming when stimuli were presented in a sequence of LH→RH than in RH→LH. These findings suggest that the intrinsic differences in lexical processing between the two hemispheres may be related to the asymmetric pattern of hemispheric interactions in visual word processing.

Left-superiority in lexical processing was maintained in sequential presentation.Recency memory of lexical processing does not lead to hemispheric asymmetry.Symmetrical pattern in intra-hemispheric repetition primings was shown.Asymmetry pattern in inter-hemispheric repetition primings was observed.Hemispheric asymmetry of lexical processing have relevance to these patterns.

Alpinumisoflavone Activates Disruption of Calcium Homeostasis, Mitochondria and Autophagosome to Suppress Development of Endometriosis.

Alpinumisoflavone is an isoflavonoid extracted from the Cudrania tricuspidate fruit and Genista pichisermolliana. It has various physiological functions, such as anti-inflammation, anti-proliferation, and apoptosis, in malignant tumors. However, the effect of alpinumisoflavone is still not known in chronic diseases and other benign reproductive diseases, such as endometriosis. In this study, we examined the cell death effects of alpinumisoflavone on the endometriosis cell lines, End1/E6E7 and VK2/E6E7. Results indicated that alpinumisoflavone inhibited cell migration and proliferation and led to cell cycle arrest, depolarization of mitochondria membrane potential, apoptosis, and disruption of calcium homeostasis in the endometriosis cell lines. However, the cellular proliferation of normal uterine epithelial cells was not changed by alpinumisoflavone. The alteration in Ca2+ levels was estimated in fluo-4 AM-stained End1/E6E7 and VK2/E6E7 cells after alpinumisoflavone treatment with or without calcium inhibitor, 2-aminoethoxydiphenyl borate (2-APB). The results indicated that a combination of alpinumisoflavone and a calcium inhibitor reduced the calcium accumulation in the cytosol of endometriosis cells. Additionally, alpinumisoflavone decreased oxidative phosphorylation (OXPHOS) in the endometriotic cells. Moreover, protein expression analysis revealed that alpinumisoflavone inactivated AKT signaling pathways, whereas it increased MAPK, ER stress, and autophagy regulatory proteins in End1/E6E7 and VK2/E6E7 cell lines. In summary, our results suggested that alpinumisoflavone could be a promising effective management agent or an adjuvant therapy for benign disease endometriosis.

Control of charge transport in electronically active systems towards integrated biomolecular circuits (IbC).

The miniaturization of traditional silicon-based electronics will soon reach its limitation as quantum tunneling and heat become serious problems at the several-nanometer scale. Crafting integrated circuits via self-assembly of electronically active molecules using a "bottom-up" paradigm provides a potential solution to these technological challenges. In particular, integrated biomolecular circuits (IbC) offer promising advantages to achieve this goal, as nature offers countless examples of functionalities entailed by self-assembly and examples of controlling charge transport at the molecular level within the self-assembled structures. To this end, the review summarizes the progress in understanding how charge transport is regulated in biosystems and the key redox-active amino acids that enable the charge transport. In addition, charge transport mechanisms at different length scales are also reviewed, offering key insights for controlling charge transport in IbC in the future.

Terbutryn causes developmental toxicity in zebrafish (Danio rerio) via apoptosis and major organ malformation in the early stages of embryogenesis.

Terbutryn (2-(ethylamino)-4-(tert-butylamino)-6-(methylthio)-1,3,5-triazine) is a substituted symmetrical triazine herbicide used in agricultural fields to prevent undesired vegetation growth by inhibiting photosynthesis in target weeds. Although terbutryn has various benefits, long-term exposure, misuse, or abuse of terbutryn may cause non-target toxicity and severe ecosystem pollution. To provide a detailed description of the embryonic developmental toxicity of terbutryn, zebrafish (Danio rerio) were exposed to 2, 4, and 6 mg/L of terbutryn and the morphological changes, pathological abnormalities, and developmental endpoints were assessed relative to that of a solvent control. The results showed that terbutryn induces a loss of survivability, reduction in body and eye size, and edema in the yolk sac. Through fluorescence microscopy, blood vessels, motor neurons, and liver development were investigated using transgenic zebrafish models based on fluorescently tagged genes (fllk1:eGFP, olig2:dsRed, and L-fabp:dsRed). Furthermore, cell death by apoptosis in zebrafish caused by terbutryn exposure was evaluated via acridine orange staining, which is a selective fluorescent staining agent. To support the preceding results, gene expression alterations caused by terbutryn exposure in zebrafish larvae were assessed. The overall results indicate that exposure to terbutryn induces apoptosis and disrupts organ development. These embryonic developmental toxicity results suggest that terbutryn should be applied in the right areas at the appropriate rates, concentrations, and quantities.

Oxyfluorfen induces cell cycle arrest by regulating MAPK, PI3K and autophagy in ruminant immortalized mammary epithelial cells.

Oxyfluorfen, a phenoxy phenyl-type herbicide, causes significant damage to ecosystems through chronically effecting invertebrates, fish, and mammals. Considering its adverse effect on ecosystem conservation, it is necessary to investigate its toxic effects on animals. However, the mechanisms of oxyfluorfen toxicity on bovines are not well established. This study investigated the cytotoxic effect of oxyfluorfen on bovine mammary epithelial cells (MAC-T). We conducted several functional experiments to examine the response of MAC-T to oxyfluorfen under various concentrations (0, 1, 2, 5, and 10 ppm). Oxyfluorfen decreased cell viability and increased apoptotic cells by regulating the expression of apoptotic genes and proteins in MAC-T. In addition, oxyfluorfen-treated cells exhibited reduced PCNA expression with a low 3D spheroid formation as compared to that of control cells. Furthermore, oxyfluorfen treatment suppressed cell cycle progression with a decrease in cyclin D1 and cyclin A2 in MAC-T. Next, we performed western blot analysis to verify intercellular signaling changes in oxyfluorfen-treated MAC-T. The phosphor-AKT protein was increased, whereas MAPK signal pathways were decreased. Particularly, the combination of oxyfluorfen with U0126 or SP600125 completely blocked the ERK1/2 and JNK pathways leading to cell viability in MAC-T. Moreover, oxyfluorfen induced inflammatory gene expression and autophagy by increasing phosphorylation of P62 and LC3B in MAC-T. These results demonstrated that oxyfluorfen has cytotoxic effect on MAC-T, implying that the milk production capacity in cows may eventually harm humans.

Tetraconazole interrupts mitochondrial function and intracellular calcium levels leading to apoptosis of bovine mammary epithelial cells.

Tetraconazole is a type of fungicide that eliminates pathogens in plants and fruit. To date, studies have focused on the direct exposure of plants and fruits to residual tetraconazole, but no studies have been conducted on the indirect effects of tetraconzaole. Given the importance of cows as milk-producing animals and their potential exposure to pesticides via plant consumption, we analyzed the mechanism by which tetraconazole influences milk production. Here, we confirmed that tetraconazole-induced apoptosis and inhibited cell viability and proliferation by regulating the cell cycle in bovine mammary epithelial cells (MAC-T). In addition, Ca2+ homeostasis in mitochondria was disrupted by tetraconazole, leading to the depolarization of mitochondrial membrane potential. Consistent with the proliferation-related findings, tetraconazole downregulated AKT, ERK1/2, P38, and JNK signaling pathways and proliferation-related proteins such as CCND1 and PCNA in MAC-T cells. Meanwhile, it upregulated cleaved caspase 3, BAX, and Cytochrome c under the same conditions in MAC-T cells. Furthermore, MAC-T exposed to tetraconazole causes a failure of proper autophagy functioning. In summary, the results of this study indicated that tetraconazole exposure may lead to a failure of milk production from bovine mammary epithelial cells by disrupting calcium homeostasis and mitochondrial function.

Oryzalin impairs maternal-fetal interaction during early pregnancy via ROS-mediated P38 MAPK/AKT and OXPHOS downregulation.

Oryzalin is a dinitroaniline pesticide for the control of weed growth via suppression of microtubule synthesis. There are studies about the deleterious effects of dinitroaniline pesticides on the reproductive system. Therefore, we attempted to demonstrate the toxic mechanisms of oryzalin on early pregnancy using porcine uterine epithelial cells (pLE) and trophectoderm (pTr) cells. According to our results, the viability and proliferation of pLE and pTr cells were suppressed in response to oryzalin exposure, and cell cycle progression was affected. Additionally, oryzalin induced apoptotic cell death and impaired mitochondrial membrane polarity in pLE and pTr cells. Moreover, we confirmed that oryzalin significantly downregulated adenosine triphosphate (ATP) production via the oxidative phosphorylation system and upregulated reactive oxygen species (ROS) generation in both pLE and pTr cells. The oryzalin-induced ROS generation was mitigated by N-acetylcysteine, a ROS scavenger, and further upregulation of phosphor-P38 MAPK/AKT/P70S6K protein expression was ameliorated in both pLE and pTr cells. We also confirmed that the suppression of migration and proliferation in oryzalin-treated pLE and pTr cells was restored upon oxidative stress mitigation. In summary, we revealed that the cytotoxic mechanisms of oryzalin-induced implantation failure were mediated by ROS-induced intracellular signaling regulation and migratory potential in pLE and pTr cells.

Melatonin inhibits endometriosis development by disrupting mitochondrial function and regulating tiRNAs.

Endometriosis is a benign gynecological disease characterized by abnormal growth of endometrial-like cells outside the uterus. Melatonin, a hormone secreted by the pineal gland, has been shown to have therapeutic effects in various diseases, including endometriosis. However, the underlying molecular mechanisms are yet to be elucidated. The results of this study demonstrated that melatonin and dienogest administration effectively reduced surgically induced endometriotic lesions in a mouse model. Melatonin suppressed proliferation, induced apoptosis, and dysregulated calcium homeostasis in endometriotic cells and primary endometriotic stromal cells. Melatonin also caused mitochondrial dysfunction by permeating through the mitochondrial membrane to disrupt redox homeostasis in the endometriotic epithelial and stromal cells. Furthermore, melatonin affected oxidative phosphorylation systems to decrease ATP production in End1/E6E7 and VK2/E6E7 cells. This was achieved through messenger RNA-mediated downregulation of respiratory complex subunits. Melatonin inhibited the PI3K/AKT and ERK1/2 pathways and the mitochondria-associated membrane axis and further suppressed the migration of endometriotic epithelial and stromal cells. Furthermore, we demonstrated that tiRNAGluCTC and tiRNAAspGTC were associated with the proliferation of endometriosis and that melatonin suppressed the expression of these tiRNAs in primary endometriotic stromal cells and lesions in a mouse model. Thus, melatonin can be used as a novel therapeutic agent to manage endometriosis.

Targeting Thymidylate Synthase and tRNA-Derived Non-Coding RNAs Improves Therapeutic Sensitivity in Colorectal Cancer.

Some colorectal cancer (CRC) patients are resistant to 5-fluorouracil (5-FU), and high expression levels of thymidylate synthase (TS) contribute to this resistance. This study investigated whether quercetin, a representative polyphenol compound, could enhance the effect of 5-FU in CRC cells. Quercetin suppressed TS levels that were increased by 5-FU in CRC cells and promoted the expression of p53. Quercetin also induced intracellular and mitochondrial reactive oxygen species (ROS) production and Ca2+ dysregulation in a 5-FU-independent pathway in CRC cells. Furthermore, quercetin decreased mitochondrial membrane potential in CRC cells and inhibited mitochondrial respiration. Moreover, quercetin regulated the expression of specific tiRNAs, including tiRNAHisGTG, and transfection of a tiRNAHisGTG mimic further enhanced the apoptotic effect of quercetin in CRC cells. An enhanced sensitivity to 5-FU was also confirmed in colitis-associated CRC mice treated with quercetin. The treatment of quercetin decreased survival rates of the CRC mouse model, with reductions in the number of tumors and in the disease activity index. Also, quercetin suppressed TS and PCNA protein expression in the distal colon tissue of CRC mice. These results suggest that quercetin has the potential to be used as an adjuvant with 5-FU for the treatment of CRC.