7-Dehydrocholesterol is an endogenous suppressor of ferroptosis.

Ferroptosis is a form of cell death that has received considerable attention not only as a means to eradicate defined tumour entities but also because it provides unforeseen insights into the metabolic adaptation that tumours exploit to counteract phospholipid oxidation1,2. Here, we identify proferroptotic activity of 7-dehydrocholesterol reductase (DHCR7) and an unexpected prosurvival function of its substrate, 7-dehydrocholesterol (7-DHC). Although previous studies suggested that high concentrations of 7-DHC are cytotoxic to developing neurons by favouring lipid peroxidation3, we now show that 7-DHC accumulation confers a robust prosurvival function in cancer cells. Because of its far superior reactivity towards peroxyl radicals, 7-DHC effectively shields (phospho)lipids from autoxidation and subsequent fragmentation. We provide validation in neuroblastoma and Burkitt's lymphoma xenografts where we demonstrate that the accumulation of 7-DHC is capable of inducing a shift towards a ferroptosis-resistant state in these tumours ultimately resulting in a more aggressive phenotype. Conclusively, our findings provide compelling evidence of a yet-unrecognized antiferroptotic activity of 7-DHC as a cell-intrinsic mechanism that could be exploited by cancer cells to escape ferroptosis.

On the Products of Cholesterol Autoxidation in Phospholipid Bilayers and the Formation of Secosterols Derived Therefrom.

In homogenous solution, cholesterol autoxidation leads to a mixture of epimers of 5 primary products, whose concentrations vary in the presence/absence of antioxidants, such as vitamin E. Two of the products (5α-OOH and 6ß-OOH) undergo Hock fragmentation to yield electrophilic secosterols implicated in disease. Herein, we show that the product distribution is similar in phospholipid bilayers, in that the 7-OOHs are the major products, but the presence/absence of vitamin E has no effect on the distribution. Cholesterol 7α-OOH, but not 7ß-OOH, undergoes Hock fragmentation to yield a mixture of unprecedented A-ring cleavage products and 6,7-epoxides. When subjected to typical derivatization conditions, 7α-OOH yields products with essentially indistinguishable chromatographic and spectroscopic features from the previously identified secosterols, casting further doubt on their controversial origin from endogenous O3 .

Pharmacokinetics and efficacy of orally administered acetaminophen (paracetamol) in adult horses with experimentally induced endotoxemia.

BACKGROUND: Acetaminophen has been evaluated in horses for treatment of musculoskeletal pain but not as an antipyretic. OBJECTIVES: To determine the pharmacokinetics and efficacy of acetaminophen compared to placebo and flunixin meglumine in adult horses with experimentally induced endotoxemia. ANIMALS: Eight university owned research horses with experimentally induced endotoxemia. METHODS: Randomized placebo controlled crossover study. Horses were treated with acetaminophen (30 mg/kg PO; APAP), flunixin meglumine (1.1 mg/kg, PO; FLU), and placebo (PO; PLAC) 2 hours after administration of LPS. Plasma APAP was analyzed via LC-MS/MS. Serial CBC, lactate, serum amyloid A, heart rate and rectal temperature were evaluated. Serum IL-1ß, IL-6, IL-8, IL-10, and TNF-α were evaluated by an equine-specific multiplex assay. RESULTS: Mean maximum plasma APAP concentration was 13.97 ± 2.74 µg/mL within 0.6 ± 0.3 hour after administration. At 4 and 6 hours after treatment, both APAP (P = <.001, P = .03, respectively) and FLU (P = .0045 and P < .001, respectively) had a significantly greater decrease in rectal temperature compared to placebo. FLU caused greater heart rate reduction than APAP at 4 and 6 hours (P = .004 and P = .04), and PLAC at 4 hours (P = .05) after treatment. CONCLUSIONS AND CLINICAL IMPORTANCE: The pharmacokinetics of acetaminophen in endotoxemic horses differ from those reported by previous studies in healthy horses. Acetaminophen is an option for antipyresis in clinical cases, particularly when administration of traditional NSAIDs is contraindicated.

Investigation of an experimental infection model of equine coronavirus in adult horses.

BACKGROUND: Equine coronavirus (ECoV) is a recently reported enteric disease of adult horses. Natural infection by ECoV has been reported in adult horses worldwide, whereas experimental infection has only been reported in juvenile horses. An experimental infection model is needed to study the clinical presentation, laboratory abnormalities, and pathophysiological changes associated with ECoV. OBJECTIVES: To investigate the clinical, hematologic, molecular, and serological features of adult horses experimentally infected with ECoV. ANIMALS: Eight adult horses. METHODS: Four horses were intragastrically infected with fecal material containing 109 genome equivalents of ECoV. Four additional horses were exposed daily to the feces from the experimentally-infected horses. Monitoring included physical examinations, as well as daily nasal swab, whole blood, and fecal collection for molecular detection of ECoV. Blood was collected every other day for hematologic analysis and weekly for serologic analysis. RESULTS: All 8 horses shed ECoV in feces. Six of the 8 horses (75%) exhibited mild, clinical disease with soft, formed manure; 1 horse exhibited transient pyrexia. All horses maintained total white cell counts within normal limits, but 3 horses developed transient lymphopenia. No statistically significant differences (P = .20) were observed in quantity of fecal shedding of ECoV between the 2 groups. CONCLUSIONS AND CLINICAL IMPORTANCE: Experimental infection of adult horses with ECoV was associated with mild and self-limiting clinical signs, transient lymphopenia, and fecal shedding of ECoV, which mimics natural infection. No differences between experimentally-infected horses and horses exposed to ECoV-containing feces were identified. Results of our study support a fecal-oral route of transmission.

Stiffness of resting lumbar myofascia in healthy young subjects quantified using a handheld myotonometer and concurrently with surface electromyography monitoring.

This study aimed to non-invasively quantify passive stiffness of superficial myofascia at a lower lumbar (L3-L4) anatomical level in young healthy male and female subjects and investigate its possible morphological variation. Resting prone lumbar myofascial measurements were quantified using MyotonPro(®) and statistically analyzed in 20 young healthy individuals over 3-weekly intervals, concurrently with surface electromyography (sEMG). Averaged mean ± SE stiffness (Newton/meter) over three weeks was significantly (p < 0.001) greater in males (247.8 ± 11.3) than females (208.4 ± 11.3), on the right (237.7 ± 12.8) than left sides (218.5 ± 12.3), at 10-min (231.4 ± 9.1) than initial baseline (224.8 ± 9.1) values. A polymorphism of stiffness values in 10 male and 10 female subjects was suggested by box plot analyses of the 3 weekly measurements and greater inter-individual than intra-individual variances. Greater knowledge of lumbar myofascial stiffness can improve understanding of their contributions in health and chronic low back disorders.

Body composition measured by dual-energy X-ray absorptiometry half-body scans in obese adults.

Dual-energy X-ray absorptiometry (DXA) has become a common measurement of human body composition. However, obese subjects have been understudied largely due to weight and scan area restrictions. Newer DXA instruments allow for heavier subjects to be supported by the DXA scanner, but the imaging area is still smaller than the body size of some obese subjects. In this study, we determined the validity of an automated half-scan methodology by comparing to the standard whole-body scans in a cohort of obese volunteers. Fifty-two subjects whose BMI>30 kg/m2 completed whole-body iDXA (GE Lunar) scans. The resulting scans were analyzed in three ways: the standard whole-body scan, total body estimated from the left side, and from the right side. Fat mass, nonbone lean mass, bone mineral content (BMC), and percent fat derived from each half scan were compared to the whole-body scans. Total fat mass, nonbone lean mass, or percent fat was comparable for the whole-body scans, left, and right side scans (>97% within individuals and >99.9% for the group). The BMC estimate using the right side scan was slightly but statistically higher than the whole-body BMC (approximately 30 g or 1%, P<0.001), while the left side scan BMC estimate was lower than the whole-body BMC by the same magnitude. No significant magnitude bias was found for any of the composition variables. We conclude that the new iDXA half-body analysis in obese subjects appears to be closely comparable to whole-body analysis for fat mass, nonbone lean mass, and percent fat.

Validity of physical activity intensity predictions by ActiGraph, Actical, and RT3 accelerometers.

OBJECTIVE: Accelerometers are promising tools for characterizing physical activity (PA) patterns in free-living persons. To date, validation of energy expenditure (EE) predictions from accelerometers has been restricted to short laboratory or simulated free-living protocols. This study seeks to determine the capabilities of eight previously published regression equations for three commercially available accelerometers to predict summary measures of daily EE. METHODS AND PROCEDURES: Study participants were outfitted with ActiGraph, Actical, and RT3 accelerometers, while measurements were simultaneously made during overnight stays in a room calorimeter, which provided minute-by-minute EE measurements, in a diverse subject population (n = 85). Regression equations for each device were used to predict the minute-by-minute metabolic equivalents (METs) along with the daily PA level (PAL). RESULTS: Two RT3 regressions and one ActiGraph regression were not significantly different from calorimeter measured PAL. When data from the entire visit were divided into four intensity categories-sedentary, light, moderate, and vigorous-significant (P < 0.001) over- and underpredictions were detected in numerous regression equations and intensity categories. DISCUSSION: Most EE prediction equations showed differences of <2% in the moderate and vigorous intensity categories. These differences, though small in magnitude, may limit the ability of these regressions to accurately characterize whether specific PA goals have been met in the field setting. New regression equations should be developed if more accurate prediction of the daily PAL or higher precision in determining the time spent in specific PA intensity categories is desired.