Anticancer Effects of Mitoquinone via Cell Cycle Arrest and Apoptosis in Canine Mammary Gland Tumor Cells.

Treating female canine mammary gland tumors is crucial owing to their propensity for rapid progression and metastasis, significantly impacting the overall health and well-being of dogs. Mitoquinone (MitoQ), an antioxidant, has shown promise in inhibiting the migration, invasion, and clonogenicity of human breast cancer cells. Thus, we investigated MitoQ's potential anticancer properties against canine mammary gland tumor cells, CMT-U27 and CF41.Mg. MitoQ markedly suppressed the proliferation and migration of both CMT-U27 and CF41.Mg cells and induced apoptotic cell death in a dose-dependent manner. Furthermore, treatment with MitoQ led to increased levels of pro-apoptotic proteins, including cleaved-caspase3, BAX, and phospho-p53. Cell cycle analysis revealed that MitoQ hindered cell progression in the G1 and S phases in CMT-U27 and CF41.Mg cells. These findings were supported using western blot analysis, demonstrating elevated levels of cleaved caspase-3, a hallmark of apoptosis, and decreased expression of cyclin-dependent kinase (CDK) 2 and cyclin D4, pivotal regulators of the cell cycle. In conclusion, MitoQ exhibits in vitro antitumor effects by inducing apoptosis and arresting the cell cycle in canine mammary gland tumors, suggesting its potential as a preventive or therapeutic agent against canine mammary cancer.

Diuron-induced fetal Leydig cell dysfunction in in vitro organ cultured fetal testes.

Diuron is a phenylurea herbicide widely used in the agricultural industry. In recent years, the risk of infertility and developmental defects has increased due to exposure to environmental pollutants. In this study, we investigated the toxicity of diuron in fetal mouse testes using three-dimensional organ cultures. Fetal testes derived from embryonic day (E) 14.5 were cultured with 200 µM diuron for 5 days. The results revealed that diuron did not impair fetal germ cell proliferation or the expression levels of germ cell markers such as Ddx4, Dazl, Oct 4, Nanog, Plzf, and TRA 98. Similarly, the gene or protein expression of the Sertoli cell markers Sox9 and Wt1 in diuron-exposed fetal testes did not change after 5 days of culture. In contrast, diuron increased fetal Leydig cell markers (FLC), Cyp11a1, Cyp17a1, Thbs2, and Pdgf α, and decreased adult Leydig cell (ALC) markers, Sult1e1, Hsd173, Ptgds, and Vcam1. However, 3-ßHSD, an FLC and ALC marker, was consistently maintained upon exposure to diuron in fetal testes compared to non-treated groups. In conclusion, our study demonstrates that diuron negatively impacts Fetal Leydig cell development, although it does not affect germ and Sertoli cells.

7-Ketocholesterol plays a key role in cholesterol-induced hepatitis via macrophage and neutrophil infiltration.

This study sought to explore the role of 7-ketocholesterol (7-KC) in liver damage caused by high cholesterol intake and its potential pathological mechanism in mice. Our in vivo findings indicated that mice fed a high-cholesterol diet had elevated serum levels of 7-KC, accompanied by liver injury and inflammation, similar to human nonalcoholic steatohepatitis. Furthermore, the high-cholesterol diet induced neutrophil infiltration, which played a critical role in liver damage through myeloperoxidase (MPO) activity. Upon stimulation with 7-KC, macrophages exhibited increased expression of C-X-C motif chemokine ligand 1 (CXCL1) and CXCL2, as well as ATP-binding cassette transporter A1 (ABCA1) and ABCG1. Hepatocytes, on the other hand, exhibited increased expression of CXCL2 and ABCG1. The infiltration of neutrophils in the liver was primarily caused by CXCL1 and CXCL2, resulting in hepatocyte cell death due to elevated MPO activity. Our data also revealed that the activation of macrophages by 7-KC via ABCA1 or ABCG1 was not associated with lipid accumulation. Collectively, these findings suggest that high cholesterol-induced hepatitis in mice involves, at least partially, the recruitment of neutrophils to the liver by 7-KC-activated macrophages. This is mediated by increased expression of CXCL1 and CXCL2 through ABCA1 or ABCG1, which act as 7-KC efflux transporters. Additionally, hepatocytes contribute to this process by increased expression of CXCL2 through ABCG1. Therefore, our findings suggest that 7-KC may play a role in high cholesterol-induced hepatitis in mice by activating macrophages and hepatocytes, ultimately leading to neutrophil infiltration.

Effect of knowledgebase transition of a clinical decision support system on medication order and alert patterns in an emergency department.

A knowledgebase (KB) transition of a clinical decision support (CDS) system occurred at the study site. The transition was made from one commercial database to another, provided by a different vendor. The change was applied to all medications in the institute. The aim of this study was to analyze the effect of KB transition on medication-related orders and alert patterns in an emergency department (ED). Data of patients, medication-related orders and alerts, and physicians in the ED from January 2018 to December 2020 were analyzed in this study. A set of definitions was set to define orders, alerts, and alert overrides. Changes in order and alert patterns before and after the conversion, which took place in May 2019, were assessed. Overall, 101,450 patients visited the ED, and 1325 physicians made 829,474 prescription orders to patients during visit and at discharge. Alert rates (alert count divided by order count) for periods A and B were 12.6% and 14.1%, and override rates (alert override count divided by alert count) were 60.8% and 67.4%, respectively. Of the 296 drugs that were used more than 100 times during each period, 64.5% of the drugs had an increase in alert rate after the transition. Changes in alert rates were tested using chi-squared test and Fisher's exact test. We found that the CDS system knowledgebase transition was associated with a significant change in alert patterns at the medication level in the ED. Careful consideration is advised when such a transition is performed.

The Anti-Inflammatory Effects of Broccoli (Brassica oleracea L. var. italica) Sprout Extract in RAW 264.7 Macrophages and a Lipopolysaccharide-Induced Liver Injury Model.

Brassica oleracea var. italica (broccoli), a member of the cabbage family, is abundant with many nutrients, including vitamins, potassium, fiber, minerals, and phytochemicals. Consequently, it has been used as a functional food additive to reduce oxidative stress and inflammatory responses. In the current study, the effects of sulforaphane-rich broccoli sprout extract (BSE) on the inflammatory response were investigated in vitro and in vivo. Comparative high-performance liquid chromatography analysis of sulforaphane content from different extracts revealed that 70% ethanolic BSE contained more sulforaphane than the other extracts. qPCR and enzyme immunoassay analyses revealed that BSE markedly reduced the expression of proinflammatory cytokines and mediators, including cyclooxygenase 2, interleukin (IL)-1ß, IL-6, IL-1, inducible nitric oxide synthase (iNOS), and tumor necrosis factor-α (TNF-α), in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Pretreatment with BSE improved the survival rate and suppressed alanine aminotransferase and aspartate aminotransferase expression in LPS-induced endotoxemic mice, while proinflammatory cytokines such as IL-1ß, TNF-α, IL-6, cyclooxygenase-2, and iNOS decreased dramatically in the LPS-induced liver injury model via BSE treatment. Additionally, F4/80 immunostaining showed that BSE suppressed hepatic macrophage infiltration in the liver after lipopolysaccharide injection. In conclusion, BSE may be a potential nutraceutical for preventing and regulating excessive immune responses in inflammatory disease.

Effects of Melatonin on Liver of D-Galactose-Induced Aged Mouse Model.

Melatonin, a hormone secreted by the pineal gland of vertebrates, regulates sleep, blood pressure, and circadian and seasonal rhythms, and acts as an antioxidant and anti-inflammatory agent. We investigated the protective effects of melatonin against markers of D-galactose (D-Gal)-induced hepatocellular aging, including liver inflammation, hepatocyte structural damage, and non-alcoholic fatty liver. Mice were divided into four groups: phosphate-buffered saline (PBS, control), D-Gal (200 mg/kg/day), melatonin (20 mg/kg), and D-Gal (200 mg/kg) and melatonin (20 mg) cotreatment. The treatments were administered once daily for eight consecutive weeks. Melatonin treatment alleviated D-Gal-induced hepatocyte impairment. The AST level was significantly increased in the D-Gal-treated groups compared to that in the control group, while the ALT level was decreased compared to the melatonin and D-Gal cotreated group. Inflammatory genes, such as IL1-ß, NF-κB, IL-6, TNFα, and iNOS, were significantly increased in the D-Gal aging model, whereas the expression levels of these genes were low in the D-Gal and melatonin cotreated group. Interestingly, the expression levels of hepatic steatosis-related genes, such as LXRα, C/EBPα, PPARα, ACC, ACOX1, and CPT-1, were markedly decreased in the D-Gal and melatonin cotreated group. These results suggest that melatonin suppresses hepatic steatosis and inflammation in a mouse model of D-Gal-induced aging.

Exploring the JAK/STAT Signaling Pathway in Hepatocellular Carcinoma: Unraveling Signaling Complexity and Therapeutic Implications.

Hepatocellular Carcinoma (HCC) continues to pose a substantial global health challenge due to its high incidence and limited therapeutic options. In recent years, the Janus Kinase (JAK) and Signal Transducer and Activator of Transcription (STAT) pathway has emerged as a critical signaling cascade in HCC pathogenesis. The review commences with an overview of the JAK/STAT pathway, delving into the dynamic interplay between the JAK/STAT pathway and its numerous upstream activators, such as cytokines and growth factors enriched in pathogenic livers afflicted with chronic inflammation and cirrhosis. This paper also elucidates how the persistent activation of JAK/STAT signaling leads to diverse oncogenic processes during hepatocarcinogenesis, including uncontrolled cell proliferation, evasion of apoptosis, and immune escape. In the context of therapeutic implications, this review summarizes recent advancements in targeting the JAK/STAT pathway for HCC treatment. Preclinical and clinical studies investigating inhibitors and modulators of JAK/STAT signaling are discussed, highlighting their potential in suppressing the deadly disease. The insights presented herein underscore the necessity for continued research into targeting the JAK/STAT signaling pathway as a promising avenue for HCC therapy.

T-2 mycotoxin Induces male germ cell apoptosis by ROS-mediated JNK/p38 MAPK pathway.

T-2 mycotoxin, a type A trichothecene toxin that, specifically, causes male and female reproductive toxicity. We evaluated T-2 toxin toxicity in testes from neonatal testes after in vitro tissue cultured. Additionally, current study focuses on the molecular mechanism of toxicity and germ cell damage in GC-1 spermatogonial cells. Mouse testicular fragments were subjected to T-2 toxin (0-20 nM) during days 5 of in vitro culture. Testicular germ cell number were reduced and downregulated the expression of corresponding markers depending on the exposure concentration of T-2 toxin; however, Sertoli cell markers and steroidogenic enzyme expression increased when treated with 20 nM T-2 toxin. The cell viability decreased, apoptosis increased, and pro-apoptotic protein expression increased in 5-20 nM T-2 toxin-exposed spermatogonia. Moreover, T-2 toxin generated reactive oxygen species (ROS) and induced mitochondrial dysfunction, indicating that activation of p38 MAPK signaling triggered by ROS is involved in the apoptotic molecular mechanism of T-2 toxin. T-2 toxin induced the phosphorylation of ERK1/2, c-Jun, JNK/SAPK, p38, and p53, and the subsequent inhibition of AKT phosphorylation. The upregulation of genes related to apoptosis and MAPK/JNK signaling was consistently observed in cells exposed to T-2 toxin. These results indicate that T-2 toxin triggers apoptotic cell death in germ cells through the triggering of ROS-mediated JNK/p38-MAPK signaling pathways.

Considering cell volume in dopant screening for improving Li-ion mobility in an amorphous LiPON solid-state electrolyte: an ab initio study.

Engineering of solid electrolytes of Li-ion batteries is carried out for achieving high levels of ionic conductivity and preserving low levels of electrical conductivity. Doping metallic elements into solid electrolyte materials composed of Li, P, and O is quite challenging due to instances of possible decomposition and secondary phase formation. To accelerate the development of high-performance solid electrolytes, predictions of thermodynamic phase stabilities and conductivities are necessary, as they would avoid the need to carry out exhaustive trial-and-error experiments. In this study, we demonstrated theoretical approach to increase the ionic conductivity of amorphous solid electrolyte by doping: cell volume-ionic conductivity relation. Using density functional theory (DFT) calculations, we examined the validity of the hypothetical principle in predicting improvements in stability and ionic conductivity with 6 candidate doping elements (Si, Ti, Sn, Zr, Ce, Ge) in a quaternary Li-P-O-N solid electrolyte system (LiPON) both in crystalline and amorphous phases. The doping of Si into LiPON (Si-LiPON) was indicated to stabilize the system and enhance ionic conductivity based on our calculated doping formation energy and cell volume change. The proposed doping strategies provide crucial guidelines for the development of solid-state electrolytes with enhanced electrochemical performances.

Tebuconazole Induces ER-Stress-Mediated Cell Death in Bovine Mammary Epithelial Cell Lines.

Tebuconazole (TEB) is a triazole fungicide used to increase crop production by controlling fungi, insects, and weeds. Despite their extensive use, people are concerned about the health risks associated with pesticides and fungicides. Numerous studies have defined the cellular toxicity of triazole groups in pesticides, but the mechanisms of TEB toxicity in bovine mammary gland epithelial cells (MAC-T cells) have not yet been studied. Damage to the mammary glands of dairy cows directly affects milk production. This study investigated the toxicological effects of TEB on MAC-T cells. We found that TEB decreases both cell viability and proliferation and activates apoptotic cell death via the upregulation of pro-apoptotic proteins, such as cleaved caspases 3 and 8 and BAX. TEB also induced endoplasmic reticulum (ER) stress via the upregulation of Bip/GRP78; PDI; ATF4; CHOP; and ERO1-Lα. We found that TEB induced mitochondria-mediated apoptotic MAC-T cell death by activating ER stress. This cell damage eventually led to a dramatic reduction in the expression levels of the milk-protein-synthesis-related genes LGB; LALA; CSN1S1; CSN1S2; and CSNK in MAC-T cells. Our data suggest that the exposure of dairy cows to TEB may negatively affect milk production by damaging the mammary glands.

Perfluorooctanoic acid induces cell death in TM3 cells via the ER stress-mitochondrial apoptosis pathway.

Perfluorooctanoic acid (PFOA) is an environmentally ubiquitous synthetic chemical highly persistent in organisms. PFOA exposure is pernicious to reproductive health as indicated by reports of male infertility. However, the PFOA toxicity mechanism to Leydig cells remains poorly understood. Therefore, this study aimed to investigate the toxicological events occurring in TM3 Leydig cells treated with PFOA (250, 500, 750 µM) for 24 h. PFOA was shown to significantly decrease cell viability resulting from inhibition of proliferation and elevation of apoptotic ratio in a dose dependent manner. Upregulation of pro-apoptotic gene expressions such as Bax, Bad, and p53, was observed in combination with an increase in the apoptosis-related protein levels of Bax, cleaved caspase-3, cleaved caspase-8, and phosphorylated p53. Furthermore, exposure of PFOA lead to mitochondrial damage involving mitochondrial membrane permeabilization. A release of cytochrome c and collapse of the mitochondrial membrane potential (∆Ψm) were observed compared to the untreated control. Additionally, PFOA stimulated unfolded protein response (UPR) upregulating ER stress marker, Bip/GRP78, and upregulated protein levels of UPR signal molecules IRE1, p-JNK, p-ERK1/2, p-p53, CHOP, and ERO1. Overall, the present study elucidated the ER stress-mitochondrial apoptosis pathway-related molecular mechanisms involved in PFOA-induced cell death in TM3 Leydig cells.

Sedative-Hypnotic Effects of Glycine max Merr. Extract and Its Active Ingredient Genistein on Electric-Shock-Induced Sleep Disturbances in Rats.

Glycine max Merr. (GM) is a functional food that provides many beneficial phytochemicals. However, scientific evidence of its antidepressive and sedative activities is scarce. The present study was designed to investigate the antidepressive and calmative effects of GM and its biologically active compound, genistein (GE), using electroencephalography (EEG) analysis in an electric foot shock (EFS)-stressed rat. The underlying neural mechanisms of their beneficial effects were determined by assessing corticotropin-releasing factor (CRF), serotonin (5-HT), and c-Fos immunoreactivity in the brain using immunohistochemical methods. In addition, the 5-HT2C receptor binding assay was performed because it is considered a major target of antidepressants and sleep aids. In the binding assay, GM displayed binding affinity to the 5-HT2C receptor (IC50 value of 14.25 ± 11.02 µg/mL). GE exhibited concentration-dependent binding affinity, resulting in the binding of GE to the 5-HT2C receptor (IC50, 77.28 ± 26.57 mg/mL). Administration of GM (400 mg/kg) increased non-rapid eye movement (NREM) sleep time. Administration of GE (30 mg/kg) decreased wake time and increased rapid eye movement (REM) and NREM sleep in EPS-stressed rats. In addition, treatment with GM and GE significantly decreased c-Fos and CRF expression in the paraventricular nucleus (PVN) and increased 5-HT levels in the dorsal raphe in the brain. Overall, these results suggest that GM and GE have antidepressant-like effects and are effective in sleep maintenance. These results will benefit researchers in developing alternatives to decrease depression and prevent sleep disorders.

The Association Between Media-Based Exposure to Nonsuicidal Self-Injury and Emergency Department Visits for Self-Harm.

OBJECTIVE: This study aimed to evaluate the association between media-based exposure to nonsuicidal self-injury (NSSI) and emergency department (ED) visits due to self-harm in Korea, specifically before and after the initial broadcast of the song Barcode, which has an explicit focus on NSSI. METHOD: We used the national emergency department information system to obtain data related to ED visits due to self-harm between January 1, 2014, and December 31, 2018. Using interrupted time series regression analysis, we assessed the monthly ED visits due to self-harm before and after the media-based exposure to NSSI on March 30, 2018. In addition, self-harm methods were assessed. RESULTS: A total of 35,928,834 visits to ED were identified, of which 115,647 were due to self-harm. ED visits due to self-harm showed a significant step increase in the 10- to 14-year-old (ß = 0.883, p = .001), 15- to 19-year-old (ß = 2.941, p < .001), 20- to 24-year-old (ß = 1.997, p = 0.002), and 25- to 29-year-old (ß = 1.438, p = .029) age groups, before and after the media-based exposure to NSSI. The most pronounced increase was observed in male participants aged 20 to 24 years (ß = 1.790, p = .012) and female participants aged 15 to 19 years (ß = 5.158, p < .001). Self-harm by cutting has increased significantly in participants aged 10 to 29 years, and self-harm by poisoning has also increased significantly in participants aged 10 to 19 and 25 to 29 years. CONCLUSION: ED visits due to self-harm increased significantly, especially in adolescents and young adults, following the exposure to NSSI. Responsible media reporting on NSSI and appropriate guidelines would help prevent a further increase in self-harm. DIVERSITY & INCLUSION STATEMENT: We worked to ensure sex and gender balance in the recruitment of human participants. We worked to ensure race, ethnic, and/or other types of diversity in the recruitment of human participants. The author list of this paper includes contributors from the location and/or community where the research was conducted who participated in the data collection, design, analysis, and/or interpretation of the work.

Deep Learning-based Approach to Predict Pulmonary Function at Chest CT.

Background Low-dose chest CT screening is recommended for smokers with the potential for lung function abnormality, but its role in predicting lung function remains unclear. Purpose To develop a deep learning algorithm to predict pulmonary function with low-dose CT images in participants using health screening services. Materials and Methods In this retrospective study, participants underwent health screening with same-day low-dose CT and pulmonary function testing with spirometry at a university affiliated tertiary referral general hospital between January 2015 and December 2018. The data set was split into a development set (model training, validation, and internal test sets) and temporally independent test set according to first visit year. A convolutional neural network was trained to predict the forced expiratory volume in the first second of expiration (FEV1) and forced vital capacity (FVC) from low-dose CT. The mean absolute error and concordance correlation coefficient (CCC) were used to evaluate agreement between spirometry as the reference standard and deep-learning prediction as the index test. FVC and FEV1 percent predicted (hereafter, FVC% and FEV1%) values less than 80% and percent of FVC exhaled in first second (hereafter, FEV1/FVC) less than 70% were used to classify participants at high risk. Results A total of 16 148 participants were included (mean age, 55 years ± 10 [SD]; 10 981 men) and divided into a development set (n = 13 428) and temporally independent test set (n = 2720). In the temporally independent test set, the mean absolute error and CCC were 0.22 L and 0.94, respectively, for FVC and 0.22 L and 0.91 for FEV1. For the prediction of the respiratory high-risk group, FVC%, FEV1%, and FEV1/FVC had respective accuracies of 89.6% (2436 of 2720 participants; 95% CI: 88.4, 90.7), 85.9% (2337 of 2720 participants; 95% CI: 84.6, 87.2), and 90.2% (2453 of 2720 participants; 95% CI: 89.1, 91.3) in the same testing data set. The sensitivities were 61.6% (242 of 393 participants; 95% CI: 59.7, 63.4), 46.9% (226 of 482 participants; 95% CI: 45.0, 48.8), and 36.1% (91 of 252 participants; 95% CI: 34.3, 37.9), respectively. Conclusion A deep learning model applied to volumetric chest CT predicted pulmonary function with relatively good performance. © RSNA, 2023 Supplemental material is available for this article.

A three-dimensional spheroid co-culture system of neurons and astrocytes derived from Alzheimer's disease patients for drug efficacy testing.

Cell culture systems derived from the progenitor cells of human patients have many advantages over animal models for therapeutic drug testing and studies of disease pathogenesis. Here we describe a three-dimensional (3D) spheroid co-culture system of neurons and astrocytes derived from induced pluripotent stem cells-neural precursor cells (iPSCs-NPCs) of Alzheimer's disease (AD) patients or healthy individuals that can provide information on drug efficacy unobtainable by 2D co-culture or monoculture approaches. iPSCs-NPCs of healthy controls or AD patients were seeded onto 96-well U-bottom plates and incubated with neuronal differentiation medium for one week and with astrocytic medium for two weeks to replicate the temporal order of cell maturation during brain development. These 3D spheroid models expressed marker proteins for mature neurons and astrocytes. In particular, patient-derived spheroids showed beta-amyloid (Aß) accumulation as revealed by thioflavin T (ThT) staining and ELISA. Aggregation of Aß induced caspase activation and cell death, while the neuroprotectants nordihydroguaiaretic acid (NDGA) and curcumin (CU) reduced the levels of both ThT and caspase staining. Taken together, these results demonstrate the feasibility of our 3D spheroids combined with ThT and caspase staining as a patient-based anti-AD drug screening platform.

Pharmacokinetic and Pharmacodynamic Modeling and Simulation Analysis of Prasugrel in Healthy Male Volunteers.

This study evaluated the pharmacokinetics and pharmacodynamics of the antiplatelet agent prasugrel, and explored its optimal dose regimens via modeling and simulation using NONMEM. We measured platelet aggregation and the serial plasma concentrations of the inactive (R-95913) and active metabolites (R-138727) of prasugrel after a single oral dose of 10-60 mg in 20 healthy adult male volunteers. A pharmacokinetic model for R-95913 and R-138727, and a pharmacodynamic model between the concentration of R-138727 and maximal platelet aggregation measured by light transmittance were constructed. The predictability of the model for platelet aggregation was evaluated by comparing the model prediction values with the observed ones not used in the construction of the model. Pharmacokinetic data were best described by a 3-compartment models for R-95913, a 1-compartment model for R-138727 with transit compartment model for absorption delay, and first-pass metabolic conversion of R-95913 into R-138727 during absorption. The association-dissociation model between R-138727 and its receptor in platelets was applied for the inhibitory effect of prasugrel on platelet aggregation. Prasugrel rapidly inhibited platelet aggregation after oral administration, with a prolonged duration of action; however, the concentration of the active metabolite decreased rapidly, which may have been due to the slow dissociation rate of R-138727 from its target receptor in platelets. The external validation suggests that our model could be used to individualize prasugrel treatment in various clinical situations. Simulation showed rapid onset of inhibitory effect with great magnitude and consistent inhibition after therapeutic dose of prasugrel.

Cisplatin Induces Apoptosis in Mouse Neonatal Testes Organ Culture.

Chemotherapy is used for childhood cancer but may lead to infertility in patients. Spermatogonia stem cells are present in the testes of prepubertal boys, although they do not produce sperm at this age. Herein, we evaluated the toxicity of cisplatin, a known medicine for cancer treatment, in neonatal mouse testes using in vitro organ culture. Mouse testicular fragments (MTFs) derived from 5.5-d postpartum mouse testes were exposed to 1-10 µg/mL cisplatin. The results showed that cisplatin significantly downregulated the expression of germ cell marker genes, including differentiated and undifferentiated, in a dose-dependent manner. In particular, a high dose of cisplatin (10 µg/mL) led to germ cell depletion. In addition, the expression levels of the Sertoli cell marker gene, the number of SOX9+ Sertoli cells, and the levels of SOX9 protein were markedly decreased in cisplatin-treated MTFs compared to controls. The mRNA expression of steroidogenic enzyme-related genes significantly increased in cisplatin-treated MTFs, except for estrogen receptor 1 (Esr1). Consistently, 3ß-hydroxysteroid dehydrogenase protein was also observed in the interstitial regions of cisplatin-treated MTFs. Altogether, our findings showed a significant impairment in germ cell development, Sertoli cell survival, and steroidogenesis in the MTFs of cisplatin-treated mice.

High Cholesterol-Induced Bone Loss Is Attenuated by Arctiin via an Action in Osteoclasts.

High cholesterol-induced bone loss is highly associated with oxidative stress, which leads to the generation of oxysterols, such as 7-ketocholesterol (7-KC). Here, we conducted in vivo and in vitro experiments to determine whether arctiin prevents high cholesterol diet-induced bone loss by decreasing oxidative stress. First, arctiin was orally administered to atherogenic diet (AD)-fed C57BL/6J male mice at a dose of 10 mg/kg for 6 weeks. Micro-computerized tomography (µCT) analysis showed that arctiin attenuated AD-induced boss loss. For our in vitro experiments, the anti-oxidant effects of arctiin were evaluated in 7-KC-stimulated osteoclasts (OCs). Arctiin decreased the number and activity of OCs and inhibited autophagy by disrupting the nuclear localization of transcription factor EB (TFEB) and downregulating the oxidized TFEB signaling pathway in OCs upon 7-KC stimulation. Furthermore, arctiin decreased the levels of reactive oxygen species (ROS) by enhancing the expression of nuclear factor erythroid 2-related factor 2 (Nrf2), catalase, and heme oxygenase 1 (HO-1), all of which affected OC differentiation. Conversely, silencing of Nrf2 or HO-1/catalase attenuated the effects of arctiin on OCs. Collectively, our findings suggested that arctiin attenuates 7-KC-induced osteoclastogenesis by increasing the expression of ROS scavenging genes in the Nrf2/HO-1/catalase signaling pathway, thereby decreasing OC autophagy. Moreover, arctiin inhibits the oxidation and nuclear localization of TFEB, thus protecting mice from AD-induced bone loss. Our findings thus demonstrate the therapeutic potential of arctiin for the prevention of cholesterol-induced bone loss.

Appropriateness of Alerts and Physicians' Responses With a Medication-Related Clinical Decision Support System: Retrospective Observational Study.

BACKGROUND: Alert fatigue is unavoidable when many irrelevant alerts are generated in response to a small number of useful alerts. It is necessary to increase the effectiveness of the clinical decision support system (CDSS) by understanding physicians' responses. OBJECTIVE: This study aimed to understand the CDSS and physicians' behavior by evaluating the clinical appropriateness of alerts and the corresponding physicians' responses in a medication-related passive alert system. METHODS: Data on medication-related orders, alerts, and patients' electronic medical records were analyzed. The analyzed data were generated between August 2019 and June 2020 while the patient was in the emergency department. We evaluated the appropriateness of alerts and physicians' responses for a subset of 382 alert cases and classified them. RESULTS: Of the 382 alert cases, only 7.3% (n=28) of the alerts were clinically appropriate. Regarding the appropriateness of the physicians' responses about the alerts, 92.4% (n=353) were deemed appropriate. In the classification of alerts, only 3.4% (n=13) of alerts were successfully triggered, and 2.1% (n=8) were inappropriate in both alert clinical relevance and physician's response. In this study, the override rate was 92.9% (n=355). CONCLUSIONS: We evaluated the appropriateness of alerts and physicians' responses through a detailed medical record review of the medication-related passive alert system. An excessive number of unnecessary alerts are generated, because the algorithm operates as a rule base without reflecting the individual condition of the patient. It is important to maximize the value of the CDSS by comprehending physicians' responses.

Mortality Risk in Pediatric Sepsis Based on C-reactive Protein and Ferritin Levels.

OBJECTIVES: Interest in using bedside C-reactive protein (CRP) and ferritin levels to identify patients with hyperinflammatory sepsis who might benefit from anti-inflammatory therapies has piqued with the COVID-19 pandemic experience. Our first objective was to identify patterns in CRP and ferritin trajectory among critically ill pediatric sepsis patients. We then examined the association between these different groups of patients in their inflammatory cytokine responses, systemic inflammation, and mortality risks. DATA SOURCES: A prospective, observational cohort study. STUDY SELECTION: Children with sepsis and organ failure in nine pediatric intensive care units in the United States. DATA EXTRACTION: Two hundred and fifty-five children were enrolled. Five distinct clinical multi-trajectory groups were identified. Plasma CRP (mg/dL), ferritin (ng/mL), and 31 cytokine levels were measured at two timepoints during sepsis (median Day 2 and Day 5). Group-based multi-trajectory models (GBMTM) identified groups of children with distinct patterns of CRP and ferritin. DATA SYNTHESIS: Group 1 had normal CRP and ferritin levels ( n = 8; 0% mortality); Group 2 had high CRP levels that became normal, with normal ferritin levels throughout ( n = 80; 5% mortality); Group 3 had high ferritin levels alone ( n = 16; 6% mortality); Group 4 had very high CRP levels, and high ferritin levels ( n = 121; 11% mortality); and Group 5 had very high CRP and very high ferritin levels ( n = 30; 40% mortality). Cytokine responses differed across the five groups, with ferritin levels correlated with macrophage inflammatory protein 1α levels and CRP levels reflective of many cytokines. CONCLUSIONS: Bedside CRP and ferritin levels can be used together to distinguish groups of children with sepsis who have different systemic inflammation cytokine responses and mortality risks. These data suggest future potential value in personalized clinical trials with specific targets for anti-inflammatory therapies.