Reversion of breast epithelial polarity alterations caused by obesity.

Molecular links between breast cancer risk factors and pro-oncogenic tissue alterations are poorly understood. The goal of this study was to characterize the impact of overweight and obesity on tissue markers of risk, using normal breast biopsies, a mouse model of diet-induced obesity, and cultured breast acini. Proliferation and alteration of epithelial polarity, both necessary for tumor initiation, were quantified by immunostaining. High BMI (>30) and elevated leptin were associated with compromised epithelial polarity whereas overweight was associated with a modest increase in proliferation in human and mice mammary glands. Human serum with unfavorable adipokine levels altered epithelial polarization of cultured acini, recapitulating the effect of leptin. Weight loss in mice led to metabolic improvements and restored epithelial polarity. In acini cultures, alteration of epithelial polarity was prevented by antioxidants and could be reverted by normalizing culture conditions. This study shows that obesity and/or dietary factors modulate tissue markers of risk. It provides a framework to set target values for metabolic improvements and to assess the efficacy of interventional studies aimed at reducing breast cancer risk.

Characterization and implementation of a miniature X-ray system for live cell microscopy.

The study of radiation effects on biological tissues is a diverse field of research with direct applications to improve human health, in particular in the contexts of radiation therapy and space exploration. Understanding the DNA damage response following radiation exposure, which is a key determinant for mutagenesis, requires reproducible methods for delivering known doses of ionizing radiation (IR) in a controlled environment. Multiple IR sources, including research X-ray and gamma-ray irradiators are routinely used in basic and translational research with cell and animal models. These systems are however not ideal when a high temporal resolution is needed, for example to study early DNA damage responses with live cell microscopy. Here, we characterize the dose rate and beam properties of a commercial, miniature, affordable, and versatile X-ray source (Mini-X). We describe how to use Mini-X on the stage of a fluorescence microscope to deliver high IR dose rates (up to 29 Gy/min) or lower dose rates (≤ 0.1 Gy/min) in live cell imaging experiments. This article provides a blueprint for radiation biology applications with high temporal resolution, with a step-by-step guide to implement a miniature X-ray system on an imaging platform, and the information needed to characterize the system.

Elevated leptin disrupts epithelial polarity and promotes premalignant alterations in the mammary gland.

Obesity is a highly prevalent and modifiable breast cancer risk factor. While the role of obesity in fueling breast cancer progression is well established, the mechanisms linking obesity to breast cancer initiation are poorly understood. A hallmark of breast cancer initiation is the disruption of apical polarity in mammary glands. Here we show that mice with diet-induced obesity display mislocalization of Par3, a regulator of cellular junctional complexes defining mammary epithelial polarity. We found that epithelial polarity loss also occurs in a 3D coculture system that combines acini with human mammary adipose tissue, and establish that a paracrine effect of the tissue adipokine leptin causes loss of polarity by overactivation of the PI3K/Akt pathway. Leptin sensitizes non-neoplastic cells to proliferative stimuli, causes mitotic spindle misalignment, and expands the pool of cells with stem/progenitor characteristics, which are early steps for cancer initiation. We also found that normal breast tissue samples with high leptin/adiponectin transcript ratio characteristic of obesity have an altered distribution of apical polarity markers. This effect is associated with increased epithelial cell layers. Our results provide a molecular basis for early alterations in epithelial architecture during obesity-mediated cancer initiation.

Radial Profile Analysis of Epithelial Polarity in Breast Acini: A Tool for Primary (Breast) Cancer Prevention.

Preventing cancer is vastly better than treating the disease in terms of a patient's quality of life and healthcare costs. Yet, to screen for chemopreventative drugs or evaluate interventions aimed at lowering cancer risk, quantitative readouts of risk are needed. In the breast and in other organs of epithelial origin, apical-basal polarity is key to homeostasis and is one of the first tissue characteristics lost during cancer initiation. Therefore, apical-basal polarity may be leveraged as an "architectural" determinant of cancer risk. A classic approach to quantify the localization of epithelial polarity markers is visual scoring at the microscope by trained investigators. This approach is time-intensive and limited to low throughput. To increase the speed, accuracy, and scoring volume, we developed an algorithm that essentially replaces the human eye to objectively quantify epithelial polarity in microscopy images of breast glandular units (acini). Acini in culture are identified based on a nuclear stain and the corresponding masks are divided into concentric terraces of equal width. This positional information is used to calculate radial intensity profiles (RP) of polarity markers. Profiles with a steep slope represent polarized structures, whereas more horizontal curves are indicative of non-polarized acini. To compare treatment effects, RP curves are integrated into summary values of polarity. We envision applications of this method for primary cancer prevention research with acini organoids, specifically (1) to screen for chemoprevention drugs, (2) for toxicological assessment of suspected carcinogens and pharmacological hit compounds, and (3) for personalized evaluation of cancer risk and risk-reducing interventions. The RadialProfiler algorithm developed for the MATLAB computing environment and for users without prior informatics knowledge is publicly available on the Open Science Framework (OSF).

Use and in-hospital mortality associated with two cardiac procedures, by sex and age: national trends, 1990-2004.

This study used data from the National Hospital Discharge Survey to examine sex- and age-specific trends in use and in-hospital mortality associated with coronary artery bypass graft (CABG) and percutaneous coronary intervention (PCI) among adults age forty-five and older during 1990-2004. Although use rates for PCI increased 58 percent over the study period, CABG use rates declined. In-hospital death rates declined or stayed the same even though comorbidities increased for patients who received the procedures. PCI and CABG use rates for men were at least twice those for women, although women generally had more comorbidities and higher in-hospital death rates.

Heart disease and prevention: race and age differences in heart disease prevention, treatment, and mortality.

OBJECTIVE: The objective of this study was to analyze race and age differences in the distribution of health promotion and cardiovascular screening tests, and the prevalence of serious heart disease and cardiovascular mortality in the United States. DATA SOURCES/STUDY POPULATION: Data are from 7 federal datasets represented in the first National Healthcare Quality Report and the National Healthcare Disparities Report, and include surveys, administrative and vital statistics data systems. The study analyzes blacks and whites. MEASURES: Counseling on diet and nutrition, exercise, and tobacco during an outpatient visit indicate the availability of health promotion services, and screening for high blood pressure and cholesterol represent preventive services. Hospitalizations for heart-related conditions and use of certain cardiac procedures identify serious heart disease. Deaths from coronary artery disease and stroke are the heart-related mortality measures. PRINCIPAL FINDINGS: Counseling and education services tend to occur more on outpatient visits by individuals aged 45 to 64 years than in younger age groups. Screening rates among individuals aged 45 to 64 years of approximately 90% for hypertension and 80% for high cholesterol suggest progress in early detection of cardiac risk factors. However, blacks aged 45 to 64 years are 5.6 times more likely than their white counterparts to be hospitalized for hypertension, approximately one third less likely to receive a cardiac procedure, and almost twice as likely to die of coronary heart disease. CONCLUSIONS: Although findings indicate few racial differences in health promotion services in ambulatory care or screening for cardiac risk factors, the prevalence of serious cardiovascular disease, use of cardiac procedures, and heart-related mortality suggest continuing racial disparities in heart disease.