Miller, Virginia M.

Calcifying nanoparticles (NPs) have been implicated in many diseases including kidney stones and vascular calcification. The chemical composition of these NPs is unknown and advances in understanding their contribution to disease etiology has been hampered by the lack of specific antibodies. Therefore, a monoclonal antibody (MoAb) was developed using decalcified NPs isolated from a human kidney stone as an antigen. After clone screening, a monoclonal antibody was produced that recognized an approximately 125kDa band in the kidney stone NP isolate. Immunoprecipitation of NP lysate followed by proteomics was used to further characterize the protein(s) recognized by the antibody. Consecutive tissue sections from a human atherosclerotic plaque were also stained with the MoAb, which recognized punctuate structures at the media/plaque border and around cholesterol clefts. The staining pattern was similar to that observed with a commercially available antibody in our previous studies. Therefore, we have developed a monoclonal antibody against human kidney stone‐derived NPs that recognizes structures in human atherosclerotic plaques. This MoAB has great potential for use in NP detection in other diverse human tissues and in experimental protocols. Supported by NIH HL88988 and Division of Experimental Surgery, Mayo Clinic.

Bone loss and development of atherosclerosis characterized by coronary arterial calcification (CAC) accelerate in women after menopause. This study evaluated the relationship between blood borne markers of bone turnover and development of CAC in newly menopausal women being screened for the Kronos Early Estrogen Prevention Study. Women (n=137) were 52.3±2.3 years of age and were 18.1±9.2 months past their last menses. CAC was measured using 64‐slice computed tomography. Concentrations of osteocalcin, bone alkaline phosphatase (BAP), tartrate‐resident acid phosphatase 5b and osteopontin as bone matrix protein in serum and plasma were evaluated by ELISA. CAC scores ranged from 0 to 327.6 Agatston Unit (AU); 113 women had scores of 0 AU, 20 had CAC scores of 0<50 and 4 had CAC scores >50. Average levels of all of the markers of bone turnover were within a normal range reported in the literature. There was a positive correlation between BAP and osteocalcin. However, levels of these markers did not correlate with age or months past menopause. None of the markers correlated with CAC scores. These results suggest that blood borne markers of bone turnover do not correlate with the presence of CAC in newly menopausal women. Additional work is needed to determine if this relationship would change with increasing age. Supported by the Kronos Longevity Research Institute and Mayo Foundation.

Cell membrane‐derived microvesicles (MV) are shed into the circulation and reflect cell‐cell interaction, intravascular and intercellular signaling process. Infection increases risk for thrombosis and LPS signaling through toll‐like receptor 4 (TLR 4 ) may activate cells to generate procoagulant (phosphatidylserine surface) MV. This study was designed to investigate MV production from leukocytes and platelets exposed to LPS in vitro . Blood from wild type (WT) and TLR 4 gene deleted (dTLR 4 ) mice was incubated with LPS (500ng/ml or 5ug/ml) for 6 hours. Flow cytometer was used to quantify MV using fluorophore conjugated cell specific antibodies plus annexin‐V. Leukocyte‐ and platelet‐derived MV showed a concentration dependent increase in the blood from WT mice; whereas in dTLR 4 mice increased counts of leukocyte‐ and platelet‐derived MV were observed only at the higher concentration of LPS. At the higher concentration of LPS, numbers of leukocyte‐derived MV increased significantly and the mean number of platelet‐derived MV was two fold greater than under control conditions. These results indicate that low concentrations of LPS act through TLR 4 but higher concentrations of LPS act through other receptors in addition to TLR 4 to increase production of leukocyte‐ and platelet‐derived MV which could account in part for increased thrombotic risk with infection. Supported by AHA‐SDG 30503Z and Mayo Foundation.

AbstractDespite significant progress in the care of patients suffering from cardiovascular disease, there remains a persistent sex disparity in the diagnosis, management, and outcomes of these patients. These sex disparities are seen across the spectrum of cardiovascular care, but, are especially pronounced in acute cardiovascular care. The spectrum of acute cardiovascular care encompasses critically ill or tenuous patients with cardiovascular conditions that require urgent or emergent decision-making and interventions. In this narrative review, the disparities in the clinical course, management, and outcomes of six commonly encountered acute cardiovascular conditions, some with a known sex-predilection will be discussed within the basis of underlying sex differences in physiology, anatomy, and pharmacology with the goal of identifying areas where improvement in clinical approaches are needed.

Background: The identification of blood-borne biomarkers for the diagnosis and prognosis of Alzheimer’s disease and related dementias is more feasible at the population level than obtaining cerebrospinal fluid or neuroimaging markers. Objective: This study determined the association of blood microvesicles, derived from cells of the neurovascular unit, with brain amyloid-β deposition in menopausal women. Methods: A subset of women from the Kronos Early Estrogen Prevention Study underwent brain amyloid-β positron emission tomography three years following cessation of study treatment with placebo (PL, n = 29), transdermal 17β-estradiol (tE2; n = 21), or oral conjugated equine estrogen (oCEE; n = 17). Isolated peripheral venous blood microvesicles were analyzed by digital flow cytometry using fluorophore conjugated antibodies directed toward total tau, amyloid-β 1–42 (Aβ1–42), neuron specific class III β-tubulin (Tuj1), microglia ionized calcium -binding adaptor molecule 1(Iba1), glial fibrillary acid protein (GFAP), and low density lipoprotein receptor-related protein1 (LRP1). Principal components analysis reduced the dimensionality of these selected six markers to two principal components (PCs). Proportional odds ordinal logistic regression analysis was used with amyloid-β deposition regressed on these PCs. Results: Only the number of microvesicles positive for Aβ1–42 differed statistically among prior treatment groups (median [IQR]: 6.06 [2.11, 12.55] in PL; 2.49 [0.73, 3.59] in tE2; and 4.96 [0.83, 10.31] in oCEE; p = 0.032). The joint association between the 2 PCs and brain amyloid-β deposition was significant (p = 0.045). Conclusion: Six selected markers expressing peripheral blood microvesicles derived from cells of the neurovascular unit, when summarized into two principal components, were associated with brain amyloid-β deposition.

Hormone therapy improves sleep in menopausal women and recent data suggest that transdermal 17β-estradiol may reduce the accumulation of cortical amyloid-β. However, how menopausal hormone therapies modify the associations of amyloid-β accumulation with sleep quality is not known. In this study, associations of sleep quality with cortical amyloid-β deposition and cognitive function were assessed in a subset of women who had participated in the Kronos early estrogen prevention study. It was a randomized, placebo-controlled trial in which recently menopausal women (age, 42–58; 5–36 months past menopause) were randomized to (1) oral conjugated equine estrogen (n = 19); (2) transdermal 17β-estradiol (tE2, n = 21); (3) placebo pills and patch (n = 32) for 4 years. Global sleep quality score was calculated using Pittsburgh sleep quality index, cortical amyloid-β deposition was measured with Pittsburgh compound-B positron emission tomography standard uptake value ratio and cognitive function was assessed in four cognitive domains 3 years after completion of trial treatments. Lower global sleep quality score (i.e., better sleep quality) correlated with lower cortical Pittsburgh compound-B standard uptake value ratio only in the tE2 group (r = 0.45, P = 0.047). Better global sleep quality also correlated with higher visual attention and executive function scores in the tE2 group (r = −0.54, P = 0.02) and in the oral conjugated equine estrogen group (r = −0.65, P = 0.005). Menopausal hormone therapies may influence the effects of sleep on cognitive function, specifically, visual attention and executive function. There also appears to be a complex relationship between sleep, menopausal hormone therapies, cortical amyloid-β accumulation and cognitive function, and tE2 formulation may modify the relationship between sleep and amyloid-β accumulation.

AbstractBackgroundThe critical‐period theory of menopausal hormone therapy (mHT) suggests that if menopausal hormone therapy begins shortly after menopause, it may have beneficial effects on brain structure and cognitive health. We investigated white matter integrity with diffusion tensor MRI (DTI) 3 years after the end of mHT trial in recently menopausal women who were randomized shortly after menopause.MethodParticipants of this study were enrolled in the Kronos Early Estrogen Prevention Study (aged 42–56 years; 5–36 months past menopause). Participants received the following treatments: 0.45 mg/d oral conjugated equine estrogen (oCEE, n=19); 50 μg/d transdermal 17β‐estradiol (tE2, n=22); or placebo (n=33) for 4 years. Oral progesterone (200 mg/d) was given to hormone therapy groups for 12 days each month. DTI was performed 3 years after the end of 4 years of hormone therapy (n=75), which corresponded to 7 years after randomization. Voxel‐based analysis was performed on DTI to evaluate fractional anisotropy (FA) and mean diffusivity (MD) differences between treatment groups (uncorrected p<0.001).ResultVoxel based analysis showed higher FA, a measure of white matter integrity, in the superior corona radiata, anterior limb of the internal capsule and splenium of the corpus callosum of participants treated with oCEE compared to placebo (p <0.001, Figure 1A). Furthermore, higher FA was observed in the superior corona radiata, anterior limb of internal capsule and thalamus in participants treated with tE2 compared to placebo (p <0.001, Figure 1B). There were no differences in FA findings between oCEE and tE2 groups, and no MD differences between treatments groups.ConclusionA relatively higher FA in a variety of white matter regions in the oCEE and tE2 groups compared to placebo suggest that mHT may have long‐term protective effects on white matter integrity. Previously, we had shown that women in the tE2 group preserved their dorsolateral prefrontal cortex volumes better than the placebo group over 7 years. Our findings suggest that the tracts that connect the dorsolateral prefrontal cortex and the thalamus such as the superior corona radiata and the anterior limb of internal capsule may particularly benefit from mHT.

Background: There are limited data on how sex influences the outcomes of acute myocardial infarction-cardiogenic shock (AMI-CS) in young adults. Methods: A retrospective cohort of AMI-CS admissions aged 18 to 55 years, during 2000 to 2017, was identified using the National Inpatient Sample. Use of coronary angiography, percutaneous coronary intervention, mechanical circulatory support and noncardiac interventions was identified. Outcomes of interest included in-hospital mortality, use of cardiac interventions, hospitalization costs, and length of stay. Results: A total 90 648 AMI-CS admissions ≤55 years of age were included, of which 26% were women. Higher rates of CS were noted in men (2.2% in 2000 to 4.8% in 2017) compared with women (2.6% in 2000 to 4.0% in 2017; P <0.001). Compared with men, women with AMI-CS were more frequently of Black race, from a lower socioeconomic status, with higher comorbidity, and admitted to rural and small hospitals (all P <0.001). Women had lower rates of ST-segment elevation presentation (73.0% versus 78.7%), acute noncardiac organ failure, cardiac arrest (34.3% versus 35.7%), and received less-frequent coronary angiography (78.3% versus 81.4%), early coronary angiography (49.2% versus 54.1%), percutaneous coronary intervention (59.2% versus 64.0%), and mechanical circulatory support (50.3% versus 59.2%; all P <0.001). Female sex was an independent predictor of in-hospital mortality (23.0% versus 21.7%; adjusted odds ratio, 1.11 [95% CI, 1.07–1.16]; P <0.001). Women had lower hospitalization costs ($156 372±$198 452 versus $167 669±$208 577; P <0.001) but comparable lengths of stay compared with men. Conclusions: In young AMI-CS admissions, women are treated less aggressively and experience higher in-hospital mortality than men.

Abstract Reporting the sex of biological material is critical for transparency and reproducibility in science. This study examined the reporting of the sex of cells used in cardiovascular studies. Articles from 16 cardiovascular journals that publish peer‐reviewed studies in cardiovascular physiology and pharmacology in the year 2018 were systematically reviewed using terms “cultured” and “cells.” Data were collected on the sex of cells, the species from which the cells were isolated, and the type of cells, and summarized as a systematic review. Sex was reported in 88 (38.6%) of the 228 studies meeting inclusion criteria. Reporting rates varied with Circulation, Cardiovascular Research and American Journal of Physiology: Heart and Circulatory Physiology having the highest rates of sex reporting (>50%). A majority of the studies used cells from male (54.5%) or both male and female animals (32.9%). Humans (31.8%), rats (20.4%), and mice (43.8%) were the most common sources for cells. Cardiac myocytes were the most commonly used cell type (37.0%). Overall reporting of sex of experimental material remains below 50% and is inconsistent among journals. Sex chromosomes in cells have the potential to affect protein expression and molecular signaling pathways and should be consistently reported.