Ferroptosis is iron-dependent, lipid peroxidation-driven, regulated cell death that is triggered when cellular glutathione peroxidase 4 (GPX4)-mediated cellular defense is insufficient to prevent pathologic accumulation of toxic lipid peroxides. Ferroptosis is implicated in various human pathologies, including neurodegeneration, chemotherapy-resistant cancers, ischemia-reperfusion injury, and acute and chronic kidney diseases. Despite the fact that the ferroptotic process has been rigorously interrogated in multiple preclinical models, the lack of specific and readily available biomarkers to detect ferroptosis in vivo in mouse models makes it challenging to delineate its contribution to key pathologic events in vivo. Critical steps to practically evaluate ferroptosis include, but are not limited to, detecting increased cell death and pathologic accumulation of toxic lipid peroxides and testing augmentation of observed pathologic events by genetic inhibition of the glutathione-GPX4 axis or mitigation of the pathologic process by ferroptosis inhibitors. Here, we describe methods to evaluate these key features of the ferroptotic process in mice in vivo. Specifically, we describe methods to detect toxic lipid peroxides (4-hydroxynonenal) and cell death (based on terminal deoxynucleotidyl transferase dUTP nick end labeling staining) as well as a protocol to pharmacologically inhibit ferroptotic stress using liproxstatin-1. These protocols provide tools for understanding the ferroptotic process in mouse genetic or disease models. © 2022 Wiley Periodicals LLC. Basic Protocol 1 How to use liproxstatin-1 Basic Protocol 2 How to evaluate ferroptosis in mouse kidneys.
Choline is an important nutrient, playing key roles in numerous metabolic pathways relevant to animal health.

The objective of this study was to evaluate the effect of dietary choline on the lipid parameters, cardiovascular health (CVH), and levels of egg trimethylamine (TMA) and cholesterol in breeder ducks during the late laying period.

A total of 60 Jingjiang ducks were randomly separated into six replicates of 10 ducks each. After peak production until 65weeks of age, the birds were fed a control basal diet. The same ducks served as the control group until 65weeks of age, when the same ducks served as the choline-supplemented group, after 15 days of dietary choline supplementation at 2955mg/kg choline above and over the basal diet initially provided. The 15 days of choline supplementation included an initial 5-day acclimatisation period.

Dietary choline supplementation increased serum TMA (p<0.01), high-density lipoprotein cholesterol, very low-density lipoprotein, and triglyceride levels in older breeder ducks. However, it did not change the levels of trimethylamine N-oxide but decreased the atherosclerosis index compared with those of the control group (p<0.01). Moreover, it increased (p<0.01) the egg yolk TMA levels but did not change the concentrations of cholesterol in egg yolk.

Dietary choline supplementation had a beneficial effect on lipid parameters and CVH in older breeder ducks, although it increased the serum and egg yolk TMA levels.
Dietary choline supplementation had a beneficial effect on lipid parameters and CVH in older breeder ducks, although it increased the serum and egg yolk TMA levels.
The purpose of this study was to evaluate pH and calcium ion release at the outer dentin surface of simulated external root resorption cavities after root canals obturated with bioceramic root canal sealer compared with those medicated with calcium hydroxide.

Sixty extracted human single-rooted teeth were selected and instrumented. External root resorption cavities were prepared at the lingual surface of the root. The teeth were randomly divided into three groups (1) Bioceramic sealer group, canals were obturated with gutta-percha and BioRoot sealer; (2) Calcium hydroxide group, canals were medicated with UltraCal XS; (3) Control group, canals were left empty. Thirty specimens were used for evaluation of pH at 7, 14, and 28 days (n = 10 per group) and the other 30 specimens were used for evaluation of calcium ion diffusion at 28 days (n = 10 per group).

Calcium hydroxide group showed the highest median pH value at all time points (7, 14, and 28 days). Both calcium hydroxide and bioceramic sealer groups showed significantly higher median pH values compared with control (p < .001). Comparing within groups, both bioceramic sealer group and calcium hydroxide group showed significantly decreased median pH over time, while the median pH of the control did not show any significant difference among Days 7, 14, and 28. Both calcium hydroxide and bioceramic sealer groups had significantly higher calcium ion release than control. Notably, bioceramic sealer group showed significantly higher calcium ion release than the calcium hydroxide group (p < .01).

Root canals obturated with gutta-percha and bioceramic sealer showed high calcium ion levels at the simulated external root resorption cavities but did not show an extended period of alkaline pH.
Root canals obturated with gutta-percha and bioceramic sealer showed high calcium ion levels at the simulated external root resorption cavities but did not show an extended period of alkaline pH.This study aimed to evaluate the bioequivalence of two pazopanib tablet formulations in healthy Chinese subjects. A randomized, open-label, single-dose, two-period, two-sequence, crossover study was conducted under fasting conditions. A total of 32 eligible subjects were randomly administered a single dose of a 200-mg generic or branded pazopanib tablet with a 16-day washout period. Blood samples were collected before and up to 72 hours after dosing. Pharmacokinetic parameters were analyzed with noncompartmental analysis. Safety assessments included physical examinations, laboratory tests, and adverse events reporting. https://www.selleckchem.com/products/ml390.html Maximum plasma concentration (Cmax ), area under the plasma concentration-time curve (AUC) from zero to the last quantifiable concentration (AUC0-t ), and AUC from zero to infinity (AUC0-∞ ) were similar between the generic and branded products (all P > .05). The 90% confidence intervals of the geometric mean ratio of the test/reference products for Cmax , AUC0-t , and AUC0-∞ were 89.1%-117.1%, 81.9%-108.5%, and 82.4%-109.6%, respectively. There were no serious adverse events during the study. The newly developed generic pazopanib tablet was bioequivalent to the reference product under fasting conditions. Both formulations were well tolerated in healthy Chinese volunteers.Extracellular vesicles (EVs) have potential biomedical applications, particularly as a means of transport for therapeutic agents. There is a need for rapid and efficient EV-liposome membrane fusion that maintains the integrity of hybrid EVs. We recently described Sf9 insect cell-derived EVs on which functional membrane proteins were presented using a baculovirus-expression system. Here, we developed hybrid EVs by membrane fusion of small liposomes and EVs equipped with baculoviral fusogenic proteins. Single-particle analysis of EV-liposome complexes revealed controlled introduction of liposome components into EVs. Our findings and methodology will support further applications of EV engineering in biomedicine.Malignant mesothelioma (MM) is a rare and highly lethal tumor that arises from mesothelial tissue on the surface of the chest and abdominal cavity. Cytological examination of body fluids, including pleural fluid and ascites, is essential for the differentiation of malignant mesothelioma from other carcinomas, such as lung and gastrointestinal carcinomas and metastatic tumors. To evaluate the effectiveness of cell block preparation procedures, which are used for immunocytochemical staining and genetic panel analysis of tumor-specific gene mutations, we used various fixatives. We also evaluated the effects of immunostaining, and the quality of nucleic acids for genetic analysis.
Cell blocks were prepared using the malignant mesothelioma cell lines MESO4 and H226 and non-small cell lung carcinoma cell line HCC78. The cells were fixed using 10% neutral buffered formalin and four different fixatives for liquid cytology. Fixed cells were formed into cell clusters using sodium alginate or centrifugation, and paraffin-embedded cell blocks were prepared.

Cell blocks were morphologically evaluated by hematoxylin and eosin and immunocytological staining, and the nucleic acid quality was evaluated by DNA/RNA extraction, qPCR, and next-generation sequence analysis. D2-40 and WT1 staining differed depending on the fixation solution and the cell cluster formation method; however, the degree of nucleic acid degradation was not impaired by any method.

Although the morphological evaluation of cytology specimens is affected by the method of cell block preparation, it is still useful for nucleic acid extraction and gene panel analysis, as long as there are sufficient amounts of tumor cells.
Although the morphological evaluation of cytology specimens is affected by the method of cell block preparation, it is still useful for nucleic acid extraction and gene panel analysis, as long as there are sufficient amounts of tumor cells.Corn stover, an underutilized agricultural residue, is a promising lignocellulosic feedstock for producing biofuels. To fully utilize it, pretreatment is needed. Typically, pretreatments are rapidly assessed using extracellular enzymes that release sugars from cellulose and hemicellulose. In contrast, this study uses methane-arrested anaerobic digestion (MAAD) to assess pretreatments. Although time consuming, MAAD is a more accurate assessment technique when lignocellulose is employed in the carboxylate platform, a promising approach that utilizes nearly all biomass components. Using recommended pretreatment conditions identified from a previous study, three corn stover pretreatments were compared using MAAD (1) shock-only, (2) NaOH-only, and (3) shock + NaOH. Air-dried sewage sludge was used as nutrient source. At 100 g/L initial substrate concentration, compared to untreated corn stover, shock-only decreased conversion (amount of biomass digested) by 14%, NaOH-only increased conversion by 82%, and shock + NaOH increased conversion by 104%. Using batch MAAD data, the continuum particle distribution model simulated four-stage countercurrent fermentation. At an industrial non-acid volatile solids (NAVS) concentration of 300 g/Lliq , for both NaOH-only and shock + NaOH, the model predicts total carboxylic acid concentration of about 58 g/L and conversion of about 0.85 g NAVSdigested /g NAVSfed at liquid retention time of 35 days and volatile solid loading rate of 4 g/(Lliq ⋅day). At this long solid residence time, shock is not necessary; however, with short solid residence times, shock acts synergistically to aid NaOH pretreatment. Shock treatment offers a way to reduce pretreatment costs without sacrificing pretreatment efficacy.Superoxide anion (O2.- ), a short-lived, highly active reactive oxygen species, participates in many physiological processes. This work reports the design of a chemiluminescent probe (CLO) based on 1,2-dioxetane-phenol with a selective and sensitive response to O2.- . The CLO consisted of a 1,2-dioxetane-phenol as a chemiluminophore core bearing a trifluoromethanesulfonate (Tf) moiety and methyl acrylate group. Upon reacting with O2.- , the Tf was specifically cleaved from the CLO, resulting in chemiluminescence generation. The CLO emits chemiluminescence at 450-650 nm (λmax =540 nm), representing visible and red chemiluminescent molecules, responsive to O2.- . The CLO processes high sensitivity (Limit of detection=66 nM) and selectivity for O2.- with and has been applied to track O2.- fluctuations in living cells and animals. In addition, CLO successfully detected and visualized O2.- -related biochemical processes, making it promising as an important imaging tool for studying redox in biology and medicine.