BRD4 inhibition sensitizes diffuse large B-cell lymphoma cells to ferroptosis.

Diffuse large B-cell lymphoma (DLBCL), the most common form of non-Hodgkin lymphoma, is characterized by an aggressive clinical course. In approximately one-third of patients with DLBCL, first-line multiagent immunochemotherapy fails to produce a durable response. Molecular heterogeneity and apoptosis resistance pose major therapeutic challenges in DLBCL treatment. To circumvent apoptosis resistance, the induction of ferroptosis might represent a promising strategy for lymphoma therapy. In this study, a compound library, targeting epigenetic modulators, was screened to identify ferroptosis-sensitizing drugs. Strikingly, bromodomain and extra-terminal domain (BET) inhibitors sensitized cells of the germinal center B-cell-like (GCB) subtype of DLBCL to ferroptosis induction and the combination of BET inhibitors with ferroptosis inducers, such as dimethyl fumarate or RSL3, synergized in the killing of DLBCL cells in vitro and in vivo. On the molecular level, the BET protein BRD4 was found to be an essential regulator of ferroptosis suppressor protein 1 expression and thus to protect GCB-DLBCL cells from ferroptosis. Collectively, we identified and characterized BRD4 as an important player in ferroptosis suppression in GCB-DLBCL and provide a rationale for the combination of BET inhibitors with ferroptosis-inducing agents as a novel therapeutic approach for DLBCL treatment.

Evaluation of the Audicor Acoustic Cardiography Device as a Diagnostic Tool in Horses with Mitral or Aortic Valve Insufficiency.

Mitral and aortic valve insufficiencies have been commonly reported in horses. The objective of this study was to establish the use of acoustic cardiography (Audicor®) in horses with aortic (AI) or mitral valve insufficiency (MI). A total of 17 healthy horses, 18 horses with AI, and 28 horses with MI were prospectively included. None of the horses was in heart failure. Echocardiography and Audicor® analyses were conducted. Electromechanical activating time (EMAT), rate-corrected EMATc, left ventricular systolic time (LVST), rate-corrected LVSTc, and intensity and persistence of the third and fourth heart sound (S3, S4) were reported by Audicor®. Graphical analysis of the three-dimensional (3D) phonocardiogram served to visually detect murmurs. Audicor® snapshot variables were compared between groups using one-way ANOVA followed by Tukey's multiple-comparisons test. The association between Audicor® snapshot variables and the corresponding echocardiographic variables was investigated by linear regression and Bland-Altman analyses. Heart murmurs were not displayed on Audicor® phonocardiograms. No significant differences were found between Audicor® variables obtained in clinically healthy horses and horses with valvular insufficiency. The Audicor® device is unable to detect heart murmurs in horses. Audicor® variables representing cardiac function are not markedly altered, and their association with corresponding echocardiographic variables is poor in horses with valvular insufficiency that are not in heart failure.

An Equine Model for Vaccination against a Hepacivirus: Insights into Host Responses to E2 Recombinant Protein Vaccination and Subsequent Equine Hepacivirus Inoculation.

Equine hepacivirus (EqHV) is the closest known genetic homologue of hepatitis C virus. An effective prophylactic vaccine is currently not available for either of these hepaciviruses. The equine as potential surrogate model for hepacivirus vaccine studies was investigated, while equine host responses following vaccination with EqHV E2 recombinant protein and subsequent EqHV inoculation were elucidated. Four ponies received prime and booster vaccinations (recombinant protein, adjuvant) four weeks apart (day -55 and -27). Two control ponies received adjuvant only. Ponies were inoculated with EqHV RNA-positive plasma on day 0. Blood samples and liver biopsies were collected over 26 weeks (day -70 to +112). Serum analyses included detection of EqHV RNA, isotypes of E2-specific immunoglobulin G (IgG), nonstructural protein 3-specific IgG, haematology, serum biochemistry, and metabolomics. Liver tissue analyses included EqHV RNA detection, RNA sequencing, histopathology, immunohistochemistry, and fluorescent in situ hybridization. Al-though vaccination did not result in complete protective immunity against experimental EqHV inoculation, the majority of vaccinated ponies cleared the serum EqHV RNA earlier than the control ponies. The majority of vaccinated ponies appeared to recover from the EqHV-associated liver insult earlier than the control ponies. The equine model shows promise as a surrogate model for future hepacivirus vaccine research.

Validation of the Accutrend Plus point-of-care triglyceride analyzer in horses, ponies, and donkeys.

OBJECTIVE: To assess the accuracy and reliability of a point-of-care (POC) triglyceride analyzer and to establish reference intervals for blood ([TRIG]POC/WB ) and plasma triglyceride concentrations ([TRIG]POC/PL ) in horses, ponies, and donkeys. DESIGN: Prospective study. SETTING: University teaching hospital. ANIMALS: 120 adult healthy equids (78 horses and ponies, 42 donkeys) and 79 equids suffering from hypertriglyceridemia (73 horses and ponies, 6 donkeys). INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: [TRIG]POC/WB and [TRIG]POC/PL were measured using a POC analyzer and plasma triglyceride concentrations were measured using a standard laboratory assay ([TRIG]LAB/PL ). Reference intervals were determined. Test accuracy was assessed by Bland-Altman comparison of POC measurements with standard laboratory measurements and by evaluating linearity of dilutional series. Test reliability was assessed by repeated serial measurements. [TRIG]POC/WB and [TRIG]POC/PL were below the analytic range of the POC assay (<0.8 mmol/L [<70 mg/dL]) in healthy horses and ponies, whereas the reference intervals were 0.82-3.14 mmol/L (73-278 mg/dL) and 0.87-3.02 mmol/L (77-267 mg/dL), respectively, in donkeys. The POC analyzer systematically overestimated triglyceride concentrations when compared to a standard laboratory assay. The difference between [TRIG]POC/WB and [TRIG]POC/PL was small and clinically negligible. The coefficient of variation of repeated measures performed on the POC analyzer was below 10% for [TRIG]POC/WB and [TRIG]POC/PL , both in horses and donkeys and at all concentration ranges. CONCLUSIONS: The POC analyzer allows accurate and reliable measurement of [TRIG]POC/WB and [TRIG]POC/PL in horses, ponies, and donkeys in clinical settings. Assay-specific reference intervals should be determined for diagnosis and clinical monitoring of hypertriglyceridemia in equids.

Assessment of method reliability and determination of reference intervals for rotational thromboelastometry in horses.

OBJECTIVES: To assess the measurement reliability of rotational thromboelastometry (ROTEM) measurements in horses, establish reference intervals for healthy horses, and evaluate the relationship between ROTEM variables, hematologic variables, and standard coagulation tests. DESIGN: Prospective observational study. SETTING: University teaching hospital. ANIMALS: Fifty healthy and 10 diseased adult horses. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Blood was sampled from 10 healthy and 10 diseased horses and samples were repeatedly analyzed to evaluate measurement reliability of various ROTEM variables. Four different ROTEM assays (ie, EXTEM, INTEM, FIBTEM, and APTEM) were run simultaneously under standardized conditions. The device-related, operator-related, and day-to-day variability for the majority of ROTEM variables was very low to low, as indicated by a coefficient of variation (CV) of < 15%. Most of test-retest variability of ROTEM variables appeared to be device-related. Blood samples from 50 clinically healthy horses were used to establish reference intervals for ROTEM variables. Multiple stepwise regression analyses identified associations of different ROTEM variables with hematocrit, total protein concentration, fibrinogen concentration, platelet count, prothrombin time, activated partial thromboplastin time, and thrombin time. CONCLUSIONS: ROTEM is a feasible method to evaluate coagulation in horses. Its measurement reliability is acceptable, but device-related measurement variability has to be considered. Reference intervals are presented, but the influence of hematocrit, platelet count, and fibrinogen concentration may need to be taken into account when interpreting individual test results.

Vascular hamartoma in the central nervous system of a foal.

Vascular hamartomas are non-neoplastic developmental anomalies of vessels. Cases of cerebral vascular hamartomas have been previously reported in dogs and cats. A 4-week-old Freiberger foal had shown persistent problems with breathing and swallowing since birth, and bilateral laryngeal paralysis was diagnosed. The foal subsequently developed left sided facial nerve paralysis and a secondary corneal ulcer in the left eye. Necropsy revealed a pinkish mass in the obex region of the brain. The mass was further investigated by histology and immunohistochemistry. Histologically, the mass consisted of many thin-walled, blood-filled vascular structures of variable diameter involving the white matter of the obex. The lining cells were immunohistochemically positive for factor VIII (von Willebrand factor) interpreted as endothelial cells. The endothelial lining showed also variable immunoreactivity for smooth muscle actin and vimentin. Normal neural parenchyma labeled with antibodies directed against glial fibrillary acidic protein and neuron-specific enolase was present between the vascular proliferations. A diagnosis of focal vascular hamartoma in the obex was made. The development of clinical signs is attributed to the compression of the surrounding neural parenchyma.