Perturbations in the uterine luminal fluid composition are detrimental to pregnancy establishment in cattle

Abstract Background A major, unresolved issue is how the uterine microenvironment determines pregnancy success in cattle. Before implantation, conceptus development depends on the uterine secretome (i.e., histotroph). Despite its pivotal role, little is known about the dynamics of histotroph synthesis and changes in composition throughout the early diestrus and the relevance to pregnancy establishment. We hypothesize that disturbances on histotroph composition affect the establishment of pregnancy. Aim was to disturb histotroph composition at early diestrus and verify the effects on: (Exp. 1) timing to restore its composition; and (Exp. 2) pregnancy rate after multiple-embryo transfer. Estrous cycle of multiparous Nelore cows were synchronized and estrus was considered d 0 (D0) of the experiments. Disturbance was through flushing each uterine horn with 30 mL of DMPBS and collecting the resulting uterine luminal flushing (ULF) on D1; D4; D7; D1 + D4 + D7. Control group remained not-collected. In Exp. 1, ULF was collected on D7.5 from all animals and used for quantification of total protein concentration and abundance of albumin. In Exp. 2, three in vitro-produced embryos were transferred to the uterine horn ipsilateral to the ovary containing the CL on D7.5 and pregnancy was checked on D25 by ultrasound. Results In Exp. 1, ULF collection on D4 or D7 increased (1.5- to 2.2-folds) the total protein concentration and albumin abundance. ULF collection on D1 did not alter (P > 0.10) these endpoints. In Exp. 2, ULF collected on D4 or D7 decreased pregnancy rates to approximately half of that measured in the remaining groups. Conclusions Subtle perturbations imposed to the native intrauterine milieu, such as those caused by a single, low-volume collection of ULF, profoundly disturbs intrauterine composition and pregnancy success. At least 4 d were necessary for the uterus to recover its composition and the functional capacity to carry post-implantation gestation

Cardiovascular Efficacy and Safety of Bococizumab in High-Risk Patients

BACKGROUN

Cardiovascular Efficacy and Safety of Bococizumab in High-Risk Patients

Bococizumab is a humanized monoclonal antibody that inhibits proprotein convertase subtilisin- kexin type 9 (PCSK9) and reduces levels of low-density lipoprotein (LDL) cholesterol. We sought to evaluate the efficacy of bococizumab in patients at high cardiovascular risk. METHODS In two parallel, multinational trials with different entry criteria for LDL cholesterol levels, we randomly assigned the 27,438 patients in the combined trials to receive bococizumab (at a dose of 150 mg) subcutaneously every 2 weeks or placebo. The primary end point was nonfatal myocardial infarction, nonfatal stroke, hospitalization for unstable angina requiring urgent revascularization, or cardiovascular death; 93% of the patients were receiving statin therapy at baseline. The trials were stopped early after the sponsor elected to discontinue the development of bococizumab owing in part to the development of high rates of antidrug antibodies, as seen in data from other studies in the program. The median follow-up was 10 months. RESULTS At 14 weeks, patients in the combined trials had a mean change from baseline in LDL cholesterol levels of -56.0% in the bococizumab group and +2.9% in the placebo group, for a between-group difference of -59.0 percentage points (P<0.001) and a median reduction from baseline of 64.2% (P<0.001). In the lower-risk, shorter-duration trial (in which the patients had a baseline LDL cholesterol level of ≥70 mg per deciliter [1.8 mmol per liter] and the median follow-up was 7 months), major cardiovascular events occurred in 173 patients each in the bococizumab group and the placebo group (hazard ratio, 0.99; 95% confidence interval [CI], 0.80 to 1.22; P = 0.94). In the higher-risk, longer-duration trial (in which the patients had a baseline LDL cholesterol level of ≥100 mg per deciliter [2.6 mmol per liter] and the median follow-up was 12 months), major cardiovascular events occurred in 179 and 224 patients, respectively (hazard ratio, 0.79; 95% CI, 0.65 to 0.97; P = 0.02). The hazard ratio for the primary end point in the combined trials was 0.88 (95% CI, 0.76 to 1.02; P = 0.08). Injection-site reactions were more common in the bococizumab group than in the placebo group (10.4% vs. 1.3%, P<0.001). CONCLUSIONS In two randomized trials comparing the PCSK9 inhibitor bococizumab with placebo, bococizumab had no benefit with respect to major adverse cardiovascular events in the trial involving lower-risk patients but did have a significant benefit in the trial involving higher-risk patients