Pathophysiology of Preeclampsia-Induced Vascular Dysfunction and Implications for Subclinical Myocardial Damage and Heart Failure.

Tragically, preeclampsia is a leading cause of pregnancy-related complications and is linked to a heightened risk for morbid and fatal cardiovascular disease (CVD) outcomes. Although the mechanism connecting preeclampsia to CVD risk has yet to be fully elucidated, evidence suggests distinct pathways of early and late preeclampsia with shared CV risk factors but with profound differences in perinatal and postpartum risk to the mother and infant. In early preeclampsia, <34 weeks of gestation, systemic vascular dysfunction contributes to near-term subclinical myocardial damage. Hypertrophy and diastolic abnormalities persist postpartum and contribute to early onset heart failure (HF). This HF risk remains elevated decades later and contributes to premature death. Black women are at the highest risk of preeclampsia and HF. These findings support closer monitoring of women postpartum, especially for those with early and severe preeclampsia to control chronic hypertension and reduce the potentially preventable sequelae of heightened CVD and HF risk.

Unregulated LDL cholesterol uptake is detrimental to breast cancer cells.

Tumor uptake of exogenous cholesterol has been associated with the proliferation of various cancers. Previously, we and others have shown that hypercholesterolemia promotes tumor growth and silencing of the LDL receptor (LDLR) in high LDLR-expressing tumors reduces growth. To advance understanding of how LDL uptake promotes tumor growth, LDLR expression was amplified in breast cancer cell lines with endogenously low LDLR expression. Murine (Mvt1) and human (MDA-MB-468) breast cancer cell lines were transduced to overexpress human LDLR (LDLROE). Successful transduction was confirmed by RNA and protein analysis. Fluorescence-labeled LDL uptake was increased in both Mvt1 and MDA-MD-468 LDLROE cells. The expression of the cholesterol-metabolizing genes, ABCA1 and ABCG1, was increased, while HMGCR was decreased in the MDA-MB-468 LDLROE cells. In contrast, Mvt1 LDLROE cells showed no differences in Abca1 and Abcg1 expression and increased Hmgcr expression. Using a Seahorse analyzer, Mvt1 LDLROE cells showed increased respiration (ATP-linked and maximal) relative to controls, while no statistically significant changes in respiration in MDA-MB-468 LDLROE cells were observed. Growth of LDLROE cells was reduced in culture and in hypercholesterolemic mice by two-fold. However, the expression of proliferation-associated markers (Ki67, PCNA and BrdU-label incorporation) was not decreased in the Mvt1 LDLROE tumors and cells. Caspase-3 cleavage, which is associated with apoptosis, was increased in both the Mvt1 and MDA-MB-468 LDLROE cells relative to controls, with the Mvt1 LDLROE cells also showing decreased phosphorylation of p44/42MAPK. Taken together, our work suggests that while additional LDL can promote tumor growth, unregulated and prolonged LDL uptake is detrimental.

TMEM176B Regulates AKT/mTOR Signaling and Tumor Growth in Triple-Negative Breast Cancer.

TMEM176B is a member of the membrane spanning 4-domains (MS4) family of transmembrane proteins, and a putative ion channel that is expressed in immune cells and certain cancers. We aimed to understand the role of TMEM176B in cancer cell signaling, gene expression, cell proliferation, and migration in vitro, as well as tumor growth in vivo. We generated breast cancer cell lines with overexpressed and silenced TMEM176B, and a therapeutic antibody targeting TMEM176B. Proliferation and migration assays were performed in vitro, and tumor growth was evaluated in vivo. We performed gene expression and Western blot analyses to identify the most differentially regulated genes and signaling pathways in cells with TMEM176B overexpression and silencing. Silencing TMEM176B or inhibiting it with a therapeutic antibody impaired cell proliferation, while overexpression increased proliferation in vitro. Syngeneic and xenograft tumor studies revealed the attenuated growth of tumors with TMEM176B gene silencing compared with controls. We found that the AKT/mTOR signaling pathway was activated or repressed in cells overexpressing or silenced for TMEM176B, respectively. Overall, our results suggest that TMEM176B expression in breast cancer cells regulates key signaling pathways and genes that contribute to cancer cell growth and progression, and is a potential target for therapeutic antibodies.

Regulation of low-density lipoprotein receptor expression in triple negative breast cancer by EGFR-MAPK signaling.

Expression of the low-density lipoprotein receptor (LDLR) has been shown to play a critical role in hypercholesterolemia-associated breast cancer growth and is associated with shorter recurrence-free survival in human breast cancer studies. We sought to identify how circulating LDL cholesterol and tumor LDLR might accelerate oncogenic processes by determining whether increased LDLR expression and cholesterol uptake are associated with the activation of the epidermal growth factor receptor (EGFR) signaling pathway in triple negative breast cancer (TNBC) cell lines. EGF stimulation of MDA-MB-468 (MDA468) cells activated p44/42MAPK (MAPK), increased expression of LDLR, and fluorescent LDL cholesterol uptake. However, stimulation of MDA-MB-231 (MDA231) cells with EGF did not lead to increased expression of LDLR despite inducing phosphorylation of EGFR. Inhibition of MAPK using UO126 in MDA231 cells reduced LDLR expression, and in MDA468 cells, UO126 impaired the LDLR increase in response to EGF. MDA468 cells exposed to the transcription inhibitor, Actinomycin, prior to treatment with EGF showed reduced degradation of LDLR mRNA compared to vehicle-treated cells. Our results suggest that the EGF-associated increase in LDLR protein expression is cell line-specific. The common pathway regulating LDLR expression was MAPK in both TNBC cell lines.

The midlife transition and the risk of cardiovascular disease and cancer Part I: magnitude and mechanisms.

Heart disease and cancer are the leading causes of death in the United States. In women, the clinical appearance of both entities-coronary heart disease and cancer (breast, endometrium, and ovary)-escalate during the decades of the midlife transition encompassing the menopause. In addition to the impact of aging, during the interval between the age of 40 and 65 years, the pathophysiologic components of metabolic syndrome also emerge and accelerate. These include visceral adiposity (measured as waist circumference), hypertension, diabetes, and dyslipidemia. Osteoporosis, osteoarthritis, sarcopenia, depression, and even cognitive decline and dementia appear, and most, if not all, are considered functionally related. Two clinical reports confirm the interaction linking the emergence of disease: endometrial cancer and metabolic syndrome. One describes the discovery of unsuspected endometrial cancer in a large series of elective hysterectomies performed in aged and metabolically susceptible populations. The other is from the Women's Health Initiative Observational Study, which found a positive interaction between endometrial cancer and metabolic syndrome regardless of the presence or absence of visceral adiposity. Both provide additional statistical support for the long-suspected causal interaction among the parallel but variable occurrence of these common entities-visceral obesity, heart disease, diabetes, cancer, and the prevalence of metabolic syndrome. Therefore, 2 critical clinical questions require analysis and answers: 1: Why do chronic diseases of adulthood-metabolic, cardiovascular, endocrine-and, in women, cancers of the breast and endometrium (tissues and tumors replete with estrogen receptors) emerge and their incidence trajectories accelerate during the postmenopausal period when little or no endogenous estradiol is available, and yet the therapeutic application of estrogen stimulates their appearance? 2: To what extent should identification of these etiologic driving forces require modification of the gynecologist's responsibilities in the care of our patients in the postreproductive decades of the female life cycle? Part l of this 2-part set of "expert reviews" defines the dimensions, gravity, and interactive synergy of each clinical challenge gynecologists face while caring for their midlife (primarily postmenopausal) patients. It describes the clinically identifiable, potentially treatable, pathogenic mechanisms driving these threats to quality of life and longevity. Part 2 (accepted, American Journal of Obstetrics & Gynecology) identifies 7 objectives of successful clinical care, offers "triage" prioritization targets, and provides feasible opportunities for insertion of primary preventive care initiatives. To implement these goals, a reprogrammed, repurposed office visit is described.

The midlife transition and the risk of cardiovascular disease and cancer Part II: strategies to maximize quality of life and limit dysfunction and disease.

Chronic dysfunction, disabilities, and complex diseases such as cardiovascular disease, diabetes mellitus type 2, osteoporosis and certain cancers, among other burdens, emerge and accelerate in midlife women. Previously in part l, we described the clinical and laboratory research findings that more readily explain and clarify the underlying pathogenetic mechanisms driving these clinical burdens, including new findings on how in particular visceral obesity and the emergence and acceleration of various components of metabolic syndrome-glucotoxicity and lipotoxicity-and a chronic systemic inflammatory state abetted by the loss of ovarian production of estradiol and the inevitable inroads of aging generate this spectrum of clinical problems. These research insights translate into opportunities for effective care strategies leading to prevention, amelioration, possible correction, and enhanced quality of life. To achieve these goals, updated detailed diagnostic, management, and therapeutic guidelines implemented by a reprogrammed and repurposed "menopause" office visit are described. A triage mechanism-when to refer to other specialists for further care-is emphasized. The previously polarized views of menopausal hormone therapy have narrowed significantly, leading to the construction of a more confident, unified, and wider clinical application. Accordingly, a menopausal hormone therapy program providing maximum benefit and minimum risk, accompanied by an algorithm for enhanced shared decision making, is included.

Investigating a Quantitative Measure of Student Self-authorship for Undergraduate Medical Education.

CONSTRUCT: In this study, the authors investigated the validity of a quantitative measure of self-authorship among medical students. Self-authorship is a cognitive-structural theory incorporating the ability to define one's beliefs, identity, and social relations to operate in a complex, ambiguous environment. BACKGROUND: Competency-based medical education (CBME) provides learners with the opportunity to self-direct their education at an appropriate pace to develop and exhibit required behaviors while incorporating functioning relationships with supervisors and trainers. Students must develop skills to adjust and succeed in this educational climate. Self-authorship is a theoretical lens that is relevant to identifying the development of the skills necessary to succeed in a CBME curriculum. Understanding the level of attained self-authorship by medical students can provide important information about which professional characteristics are more prevalent among those who are more self-authored and about how students succeed in medical school. Although there are calls in the extant literature for the application of self-authorship in medical education, there is no quantitative measure to assess its development among medical students. APPROACH: The authors developed a survey to measure self-authorship, including a free text question regarding the thought process around a hypothetical ethical situation during training. Data were collected in 2014 and 2015 from undergraduate medical students and analyzed using factor analysis and qualitative analysis of the free text. Validity evidence was sought regarding content, internal structure, and relationships to other variables. RESULTS: Analysis supports the use of a 22-item instrument to assess 3 constructs of self-authorship: asserting independence and autonomy, knowledge processing, and sense of self in ethical situations. Content analysis of text responses supported the ability of the instrument to separate development, or a lack thereof, of self-authorship. CONCLUSIONS: The authors identified an instrument that measures multidimensional, higher-order characteristics that intersect with self-authorship. This instrument can be useful at a macro level for curricular and student assessment of self-authorship. Development of these characteristics can help foster success in a CBME environment and support curricular efforts in this regard. Understanding a student's level of self-authorship can help identify areas for support as well as allow for comparisons of different student characteristics.

Aging, the menopausal transition, and hormone replenishment therapy: retrieval of confidence and compliance.

The negative health consequences of age and the postmenopausal hypoestrogenic state and their clinical management are described in this paper. While some dysfunctional elements are irreversibly entrained, others can be modified by an updated menopausal hormone therapy (MHT) strategy reflecting revision and reassessment of the initial Women's Health Initiative (WHI) reports. However, until that positive outcome is realized, menopausal women and their healthcare providers must re-establish willingness to utilize these effective strategies. This reluctance is fueled by persistent mistaken application of WHI results that reflect the bundled risk and benefits of MHT in asymptomatic women aged 50-79 (average age 63), as opposed to women in their 40s and 50s who initiate therapy because of vasomotor symptoms. The absolute risk of adverse outcomes is much lower and the benefits dramatically higher in these younger women. In addition, newer FDA-approved formulations, lower dosage regimens, and transdermal delivery methodologies offer wider choice of approaches and promise even greater safety. The positive and negative lessons of the WHI have been learned and can be applied effectively.

Intrauterine Reprogramming of the Polycystic Ovary Syndrome: Evidence from a Pilot Study of Cord Blood Global Methylation Analysis.

Polycystic ovary syndrome (PCOS) affects 5-15% of women. PCOS is a heterogeneous disorder displaying endocrine, metabolic, and reproductive dysfunction and cardiovascular risk manifestations. Evidence of heritability exists, but only a portion of the genetic transmission has been identified by genome-wide association studies and linkage studies, suggesting epigenetic phenomena may play a role. Evidence implicates intrauterine influences in the genesis of PCOS. This was a pilot study that aimed at identifying an epigenetic PCOS reprogramming signature by profiling the methylation of the DNA extracted from umbilical cord blood (UCB) from 12 subjects undergoing in vitro fertilization. Six subjects were anovulatory PCOS women diagnosed by Rotterdam criteria and six ovulatory non-PCOS women matched for age and body mass index. UCB was collected at delivery of the placenta; the DNA was extracted and submitted to methylation analysis. A differential methylation picture of prevalent hypomethylation affecting 918 genes was detected. Of these, 595 genes (64.8%) carried single or multiple hypomethylated CpG dinucleotides and 323 genes (35.2%) single or multiple hypermethylated CpG dinucleotides. The Ingenuity Pathway Analysis (IPA) online platform enlisted 908 of the 918 input genes and clustered 794 of them into 21 gene networks. Key features of the primary networks scored by IPA included carbohydrate and lipid metabolism, neurotransmitter signaling, cardiovascular system development and function, glycosaminoglycan signaling regulation and control of amino acid biosynthesis. Central to the network activities were genes controlling hormonal regulation (ESR1), mitochondrial activity (APP, PARK2), and glucose metabolism (INS). Regulatory pathways such as G-protein coupled receptor signaling, inositol metabolism, and inflammatory response were also highlighted. These data suggested the existence of a putative "PCOS epigenomic superpathway" with three main components: glucotoxic, lipotoxic, and inflammatory. If our results are confirmed, they hint at an epigenetic at risk PCOS "signature" may thus exist that may be identifiable at birth. Additional studies are needed to confirm the results of this pilot study.

Competencies as the basis for reformed premedical education. The case for an unrestricted liberal arts collegiate education.

As the HHMI-AAMC declared, their report should be taken as a "first step in a continuing conversation about the appropriate skills and knowledge," and, echoing the ACGME and GPEP, "values and attitudes that future physicians should possess." (9pExecSum) As a new formulation evolves, the premedical curriculum must foster "scholastic vigor, analytic thinking, quantitative assessment and analysis of complex systems." (9pExecSum) Based on the Mount Sinai experience, these qualities are not engendered solely nor confined to engagement in natural sciences. Students involved in a variety of baccalaureate liberal arts endeavors appear to acquire similar intellectual competencies. Furthermore, when performed successfully in challenging collegiate environments, a thorough liberal arts education may yield precisely the same values, attitudes, and behavioral characteristics all agree are essential to the medical profession and preparing physicians for the twenty-first century.

Challenging traditional premedical requirements as predictors of success in medical school: the Mount Sinai School of Medicine Humanities and Medicine Program.

PURPOSE: Students compete aggressively as they prepare for the MCAT and fulfill traditional premedical requirements that have uncertain educational value for medical and scientific careers and limit the scope of their liberal arts and biomedical education. This study assessed the medical school performance of humanities and social science majors who omitted organic chemistry, physics, and calculus, and did not take the MCAT. METHOD: The authors compared and contrasted the academic outcomes of 85 Humanities and Medicine Program (HuMed) students at Mount Sinai School of Medicine with those of their 606 traditionally prepared classmates for the 2004-2009 graduating classes. The authors analyzed basic science knowledge, clerkship performance, humanism, leadership, community service, research fellowships, distinctions, and honors. RESULTS: There were no statistically significant differences between the groups in clerkship honors other than psychiatry (HuMed students outperformed their peers, P < .0001) or in commencement distinctions or honors. Although HuMed students were significantly more likely to secure a scholarly-year mentored project (P = .001), there was no difference in graduating with distinction in research (P = .281). HuMed students were more likely to have lower United States Medical Licensing Examination Step 1 scores (221 +/- 20 versus 227 +/- 19, P = .0039) and to take a nonscholarly leave of absence (P = .0001). There was a trend among HuMed students toward residencies in primary care and psychiatry and away from surgical subspecialties and anesthesiology. CONCLUSIONS: Students without the traditional premedical preparation performed at a level equivalent to their premedical classmates.

Impact of hormone therapy for women aged 35 to 65 years, from contraception to hormone replacement.

BACKGROUND: Midlife women (aged 35-65 years) present a complex combination of clinical challenges and health care opportunities. To meet these issues effectively, recognition of the various phases of the entire menopausal transition is necessary, because each possesses unique biological properties underlying phase-specific clinical presentations. OBJECTIVE: The aim of this article is to inform health care decisions by defining the endocrine, metabolic, and clinical consequences of therapeutic inaction or intervention at each stage of the midlife experience. METHODS: Using PubMed, MEDLINE was searched for age- and phase-specific publications about ovarian function and corresponding clinical manifestations in women aged 35 to 65 years. Large, long-term longitudinal prospective, case-control, and observational studies were selected for inclusion. Results of the Framingham Heart Study, Study of Women's Health Across the Nation, Nurses' Health Study (NHS), and Women's Health Initiative (WHI), as well as materials from the World Health Organization and American College of Obstetricians and Gynecologists, were obtained from the relevant groups' Web sites in 2008. RESULTS: Synthesis of the data acquired, particularly the confirmatory and contrasting elements displayed in the WHI and NHS publications, leads to a set of guiding principles whereby individualized phase-specific management strategies may be safely employed. These include the value of weight control and exercise; use of specific nonhormonal therapies for defined indications; definition of strict inclusion/exclusion criteria; and individualization of timing, regimen, dosage, and portal of entry for possible hormone therapy. CONCLUSION: An evidence-based, restrictive inclusion/exclusion strategy can be used to maximize benefits and minimize risks for this large, growing, and health-conscious but increasingly vulnerable population.

Time and the risk of ruptured tubal pregnancy.

OBJECTIVE: Tubal pregnancy remains an important cause of maternal morbidity and mortality. We sought to quantify the relationship of time between symptom onset and treatment and the risk of tubal rupture. METHODS: We reviewed inpatient, clinic, and physician office charts of 221 women with tubal pregnancy. We assessed the conditional risk of rupture with passing time and other factors related to rupture. RESULTS: Time between symptom onset and treatment varied from 3 hours to 66 days with an average of 7 days. There was a 32% rupture rate. The conditional risk of rupture was highest within the first 48 hours of symptom onset (5-7%). The risk dropped, leveled off, and remained fairly steady at approximately 2.5% per 24 hours of untreated symptoms. Classic tubal pregnancy signs, symptoms, and tests were not helpful in predicting rupture. CONCLUSION: The rate of rupture is highest in women with the shortest times between symptom onset and treatment. With passing time, the risk declines, but remains steady despite women's getting into care. LEVEL OF EVIDENCE: II-2