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BACKGROUND Reduction of cumulative exposure to endogenous ovarian steroid hormones is a postulated method for reducing the risk of carcinoma of the breast and other malignancies. Although there are data from trials evaluating the effect of low-fat and high-fiber diets on sex hormone levels in premenopausal women, to the authors' knowledge none of these trials has combined a relatively large number of participants, follow-up of > 2–3 months, parallel controls receiving a usual diet, and careful timing of blood sampling within the menstrual cycle. METHODS A total of 213 healthy women, ages 20–40 years, were randomly assigned to follow their usual diet or to adopt an isocaloric diet with goals of 20% calories as fat, total fiber of 25 g/day, and at least 8 fruit or vegetable servings per day. Serum levels of total estradiol (E2), sex hormone-binding globulin (SHBG), non-SHBG-bound estradiol (NSBE2), SHBG, and progesterone were evaluated during a menstrual cycle at baseline, and at 4 cycles (C4) and 12 cycles (C12) after the start of the intervention. Serum was collected during each test cycle 7–9 days after the detection of an luteinizing hormone peak in the urine. One hundred eighty-nine women provided serum at C4 and 176 women at C12. RESULTS Serum E2 decreased by an average of 7.5% or 7.8 pg/mL (95% confidence interval [95% CI], −16.0–0.04) at C12 in the intervention group, versus a decrease of 0.9% or 0.9 pg/mL (95% CI, −9.5–7.7) in the control group (the P value for the difference between the treatment groups was 0.25). Results for NSBE2 were very similar to those for total estradiol. There were no material effects found to result from intervention with regard to SHBG or progesterone. The results did not differ by baseline age, body mass index, or baseline hormone level above or below the median, and were not likely to be affected by weight change, which amounted to a mean loss of only 0.23 kg in the diet group versus a gain of 0.17 kg in the control group. The decrease in serum E2 associated with intervention was not greater when subjects were stratified by self-reported adherence to the dietary goals. CONCLUSIONS The results of the current study suggest that the effects of this isocaloric low-fat, high-fiber diet pattern on circulating ovarian steroids were modest or nonexistent. However, the observed 7.5% reduction in estradiol could have biologic significance if it persisted over many years. Moreover, underestimation of the true dietary effect could have occurred because of incomplete adherence to assigned diets. Weight loss and weight control through midlife could be a more effective and feasible approach to dietary intervention in reducing the risk of breast carcinoma. Cancer 2003. © 2003 American Cancer Society.

The study was designed to determine the process and limitations by which estrone sulfate may be a precursor of estradiol in the parenchymal cells of the normal breast. The concentration of estrone sulfate in breast nipple aspirate fluid was 1000-fold greater than that of estradiol. Concentrations of 3H-estrone sulfate in parenchymal cells were only 0.20-0.33 times that of the 1.0 nM concentration in the medium, while 3H-estrone achieved concentrations up to 24 times that in the medium at 37 degrees C. Nevertheless, estrone sulfate added to the medium was linearly converted within a 1000-fold concentration range to estrone in intact cells with a mean half-time of conversion of 628 min per 10(6) cells. Homogenized cells had a half-time of 246 min per 10(6) cells. Thus, the time for entry of estrone sulfate into cells reduced the rate by approximately 55%. In split samples, the Vmax values (+/- S.D.) for intact and homogenized cells were 12.6 +/- 1.4 and 18.3 nmol/h mg DNA, respectively (P

BACKGROUND Plasma levels of insulin-like growth factor-I (IGF-I) have been associated with the risk of prostate cancer. However, the association of IGF-I with specific tumor stage and grade at diagnosis, which correlate with risk of recurrence and mortality, has not been studied rigorously. To determine whether plasma levels of IGF-I and its main circulating binding protein, IGF binding protein-3 (IGFBP-3), predict more aggressive forms of prostate cancer, we investigated the association between plasma levels of each and specific stages and grades of prostate cancer. METHODS We examined 530 case patients and 534 control subjects in a nested case-control study in the prospective Physicians' Health Study. Patients with prostate cancer diagnosed from 1982 through 1995 were matched to control subjects by age and smoking status. IGF-I and IGFBP-3 were measured in prospectively collected plasma samples. Conditional logistic regression models were used to estimate the relative risks (RRs) for prostate cancer associated with IGF-I and IGFBP-3, stratified on grade (Gleason score > or = 7 versus

Purpose: Determining total prostate specific antigen (PSA) in plasma can often identify men who are subsequently diagnosed with prostate cancer. However, excess false-positives create large financial and psychological burdens in prostate cancer screening. The percent free PSA is lower when prostate cancer is present, although to our knowledge no large prospective analyses to date have evaluated whether adding testing for free PSA may decrease false-positives, while maintaining or perhaps improving the detection of potentially curable tumors.Materials and Methods: We measured total and percent free PSA in banked plasma samples from the Physicians’ Health Study in 430 men who were later diagnosed with prostate cancer and 1,642 age matched controls who were not diagnosed with prostate cancer during a 12-year observation period. We calculated the number of cancers detected and the number of false-positives for various strategies of combined free and total PSA levels, and compared them to the use of total PSA ...

Purpose Mutations in BRCA1 are associated with familial as well as sporadic aggressive subtypes of breast cancer, but less is known about whether BRCA1 expression or subcellular localization contributes to progression in population-based settings. Methods We examined BRCA1 expression and subcellular localization in invasive breast cancer tissues from an ethnically diverse sample of 286 patients and 36 normal breast tissue controls. Two different methods were used to label breast cancer tissues for BRCA1: (1) Dual immunofluoresent staining with BRCA1 and cytokeratin 8/18 and (2) immunohistochemical staining using the previously validated MS110 mouse monoclonal antibody. Slides were visualized and quantified using the VECTRA Automated Multispectral Image Analysis System and InForm software. Results BRCA1 staining was more intense in normal than in invasive breast tissue for both cytoplasmic (p

Background: Ambient air metal concentrations have recently been implicated in the etiology of breast cancer. Previous studies have shown airborne concentrations of arsenic and cadmium are associated with the development of estrogen receptor-negative tumors. This study aims to replicate these findings and examine the role of other toxic and essential heavy metals. Methods: Participants were women who were diagnosed with breast cancer in Chicago between 2005 and 2008. We examined estrogen and progesterone receptor-negative (ER-/PR-) and high-grade tumors as markers of aggressive breast cancer, and estimated 15-year markers of exposure of 11 heavy metals. Exposures were calculated using census tract-level airborne concentrations from the National-scale Air Toxics Assessment and participants’ residential histories. We adjusted all models for socioeconomic status and reproductive factors. Results: We found that prevalent ER-/PR- tumors were associated with increased residential exposure to airborne concentrations of antimony (Q4 vs Q1: OR= 1.81; 95% CI: 0.95, 3.44; Ptrend= 0.04), cobalt (Q4 vs Q1: OR= 2.37; 95% CI: 1.26, 4.45; Ptrend < 0.01), manganese (Q4 vs Q1: OR= 2.55; 95% CI: 1.24, 5.24; Ptrend= 0.04), and selenium (Q4 vs Q1: OR= 1.85; 95% CI: 1.03, 3.29; Ptrend= 0.05), and also identified marginally significant trends for arsenic (Ptrend= 0.06), chromium (Ptrend= 0.08), lead (Ptrend= 0.08), and mercury (Ptrend= 0.07). We did not identify any overall associations with high-grade tumors, however when stratifying by menopausal status we found that antimony (Q4 vs Q1: OR= 6.97, 95% CI: 1.61-30.19) and arsenic (Q4 vs Q1: OR= 6.97, 95% CI: 1.61-30.19) were associated with prevalent high-grade tumors in premenopausal women only. Discussion: This study found further support for a role of airborne arsenic concentrations, and novel evidence implicating other airborne estrogen-pathway disrupting metal concentrations, in the development of aggressive breast cancer subtypes. Additionally, this is the first study to implicate heavy metal exposure in the etiology of high-grade tumors. These results suggest that long-term, low-dose exposures to certain heavy metals play a role in the etiology of aggressive breast cancer characteristics. Airborne exposures have the ability to affect large populations and findings from this and similar studies have large public health implications. Citation Format: Jacob K. Kresovich, Serap Erdal, Maria Argos, Hua Yun Chen, Peter H. Gann, Garth H. Rauscher. Residential airborne heavy metal concentrations and breast cancer characteristics [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2305. doi:10.1158/1538-7445.AM2017-2305

Imatinib inhibits the ABL tyrosine kinase and is effective for the treatment of chronic myeloid leukemia (CML). ABL activates GPx-1, an enzyme associated with protection against oxidative DNA damage and disease. Enzyme activity was assessed in sample pairs consisting of mononuclear cells obtained from patients before and after imatinib therapy. Control sample sets obtained from patients not receiving imatinib showed little change in GPx activity over a several month interval. Five of 7 sample sets obtained from imatinib-receiving patients showed changes in GPx activity greater than 30%. One sample decreased 42% while 4 others increased 33% to 208%. Patients with the largest increase in activity were female and had the lowest baseline levels of GPx activity. Changes in GPx activity may influence the clinical outcome of patients being treated for CML.

We have some concerns about the results presented in a paper by Loud et al. [1]. Although specimens from the 30 subjects in their study without BRCA1/2 mutations could not be measured by their method, subjects with these mutations had an unconjugated estradiol (E2) serum versus nipple aspirate fluid (NAF) concentration correlation of 0.77 and a corresponding correlation of 0.56 for E2 between serum and ductal lavage supernatant (DLS). Our concern is about their ability to measure any of the estrogens based on the method as described in the paper. The authors state in their earlier methods paper by Xu et al. [2], that the limit of quantitation of their method is 400 fg (1.3–1.5 fmol). These are on-column values comparable to those that we have found [3]. Loud et al. [1] also report a limit of detection of 10 fg on column using a capillary HPLC, which is not unreasonable. In the Methods section of the paper, they state that they collected up to 10 ll of NAF, diluted it to 1.0 ml, and split the samples into two aliquots of 0.4 ml each for total and unconjugated estrogens. Each of these was processed, derivatized, and reconstituted to 64 ll. One-eighth (equivalent to 8 ll) of the samples were injected into the HPLC. Thus, only 0.5 ll of the initial NAF sample was injected. With the reported geometric mean concentration of unconjugated estradiol at 4.21 pmol/l (0.00421 fmol/ll) (Table 3), the injected 0.5 ll contained only 0.0021 fmol or 0.6 fg (using MW of *280). Thus, they appear to be measuring 6 % of the LOD on average. It is not possible to quantify such data reliably. Earlier in the paper (p 520), they state that ‘‘For purposes of this study, all BRCA1 or BRCA2 mutation positive postmenopausal women with sufficient volumes of serum, NAF and/or DLS specimens (C0.5 ml) were eligible for inclusion.’’ If this was the actual volume of NAF extracted, this would bring the average mass of estradiol injected to 29.4 fg, well within the range of the assay, although values less than 34 % of the mean would still be below the LOD. If this was the volume of NAF obtained, more information is needed to describe subjects that produced this volume of NAF, which in our experience is a very large yield. The data on DLS are also a problem. The stated concentrations are in the same range as the NAF values, but there is a question as to how the volume of breast fluid in the lavage was determined.

Estradiol (E2) in nipple aspirate fluid (NAF), ductal lavage fluid (DLF), and random fine needle aspirates (rFNA) are compared. Quantification was by immunoassay or tandem MS. The percent of women yielding NAF varied between 24% and 48% and for DLF was 86.3%. Variation between ducts within a breast was not less than variation between breasts within women but variation between breasts and within women over time was significantly less than variation between women. Serum E2 was highly significantly different among phases of the menstrual cycle but NAF E2 was not different. The correlation between serum and breast fluid E2 concentrations in premenopausal women had coefficients of determination of less than 15%. The correlation between serum and NAF in studies of postmenopausal women varied greatly and may depend on patient selection. The difference between NAF E2 between pre- and postmenopausal women was only 22%; for rFNA it was non-significantly 44% lower in a similar group of postmenopausal women. Progesterone was 96% and 98% lower in postmenopausal NAF and rFNA samples, respectively. Measurements of E2 in breast fluid or breast tissue appears to provide similar estimates of E2 exposure. E2 levels in breast fluid do not reflect the rapid changes that occur in serum and, thus, serum availability of E2 is only one factor determining its levels in the breast. The similarity of levels between breasts and between ducts suggests that estimates of estrogen exposure does not require multiple samples, however, unavailability of fluid may require rFNA in some cases.

Background The effect of prostate-specific antigen (PSA) screening on prostate cancer mortality remains debated, despite evidence from randomized trials. We investigated the association between prostate cancer incidence, reflecting uptake of PSA testing, and prostate cancer mortality. Methods The study population consisted of all men aged 50 to 74 years residing in eight counties in Sweden with an early increase in prostate cancer incidence and six counties with a late increase during two time periods. Incidence of metastatic prostate cancer was investigated in the period from 2000 to 2009, and prostate cancer–specific mortality and excess mortality were investigated in the period from 1990 to 1999 and the period from 2000 to 2009 by calculating rate ratios for high- vs low-incidence counties and rate ratios for the period from 2000 to 2009 vs the period from 1990 to 1999 within these two groups. All statistical tests were two-sided. Results There were 4528134 person-years at risk, 1577 deaths from prostate cancer, and 1210 excess deaths in men with prostate cancer in high-incidence counties and 2471373 person-years at risk, 985 prostate cancer deaths, and 878 excess deaths in low-incidence counties in the period from 2000 to 2009. Rate ratios in counties with high vs low incidence adjusted for time period were 0.81 (95% confidence interval [CI] = 0.73 to 0.90) for prostate cancer– specific mortality and 0.74 (95% CI = 0.64 to 0.86) for excess mortality, and the rate ratio of metastatic prostate cancer was 0.85 (95% CI = 0.79 to 0.92). Conclusions The lower prostate cancer mortality in high-incidence counties reflecting a high PSA uptake suggests that more-intense as compared with less-intense opportunistic PSA screening reduces prostate cancer mortality.

A high calcium intake, mainly from dairy products, may increase prostate cancer risk by lowering concentrations of 1,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)], a hormone thought to protect against prostate cancer. The results of epidemiologic studies of this hypothesis are inconclusive.We investigated the association between dairy product and calcium intakes and prostate cancer risk in the Physicians' Health Study, a cohort of male US physicians.At baseline, the men answered abbreviated dietary questionnaires. During 11 y of follow-up, we documented 1012 incident cases of prostate cancer among 20885 men. We estimated dairy calcium intake on the basis of consumption of 5 major dairy products and used logistic regression to estimate relative risk.At baseline, men who consumed600 mg Ca/d from skim milk had lower plasma 1,25(OH)(2)D(3) concentrations than did those consumingor =150 mg Ca/d [71 compared with 85 pmol/L (30.06 compared with 35.64 pg/mL); P = 0.005]. Compared with men consumingor =0.5 daily servings of dairy products, those consuming2.5 servings had a multivariate relative risk of prostate cancer of 1.34 (95% CI: 1.04, 1.71) after adjustment for baseline age, body mass index, smoking, exercise, and randomized treatment assignment in the original placebo-controlled trial. Compared with men consumingor =150 mg Ca/d from dairy products, men consuming600 mg/d had a 32% higher risk of prostate cancer (95% CI: 1.08, 1.63).These results support the hypothesis that dairy products and calcium are associated with a greater risk of prostate cancer.

Cell growth in the human prostate is largely controlled by peptide growth factors acting through autocrine, paracrine, and even intracrine mechanisms. These growth factors respond to endocrine hormone signals and are capable of regulating proliferation, apoptosis, differentiation, and interaction with the extracellular matrix (1, 2). Early in cancer development, imbalances in stimulatory and inhibitory signals could create a "field effect", in which hyperproliferation leads to somatic mutation and clonal selection of malignant phenotypes. Measurement of growth factors and understanding of the environmental and genetic factors that upor down-regulate their expression (or the expression of their receptors), therefore, should be of great value to investigators interested in the etiology and prevention of prostate cancer. However, because these growth factors operate in the local microenvironment, it is not easy to develop intermediate biomarker assays in accessible biologic fluids or tissues suitable for population studies. The purpose of this presentation is to review the rationale for measuring specific growth factors, the various methods that have been attempted to date, and the challenges currently faced in this area of research. Valid biomarkers of growth factor expression would have several major applications in the domain of prostate cancer epidemiology and prevention. First, growth factor levels could provide important intermediate biomarkers in trials of lifestyle and chemopreventive agents. Second, growth factor patterns could predict the aggressiveness of prostate cancers before surgery—a crucial point given the variability in the behavior of prostate tumors and the number of men now diagnosed after prostate-specific antigen screening. Third, growth factor levels could clarify the prognosis in cases of resected prostate cancer. And fourth, growth factor levels could provide insights into the molecular etiology of

Objective: Findings from epidemiologic studies on the association between diabetes and prostate cancer risk are inconsistent. However, data from at least three studies suggest that the direction and strength of this association differs according to duration of diabetes. To determine the potential effects of early-stage abnormal glucose metabolism on risk, we assessed the relationship of postload glycemia in the absence of self-reported diabetes with risk of prostate cancer mortality.

Initial results of the REDUCE trial suggest that dutasteride decreases the incidence of prostate cancer in men with persistently elevated PSA; however, questions have emerged about the drug's effect on the incidence of high-grade cancer. Adjustment for post-randomization confounders, however, brings the true benefit of this drug into clearer focus.

BACKGROUND Although axillary lymph node metastasis is one of the most important prognostic determinants of breast carcinoma prognoses, the reasons why tumors vary in their capability to produce for axillary metastases remain unclear. METHODS The authors used data from the nationwide Patient Care Evaluation (PCE) survey of the American College of Surgeons to evaluate the correlations between patient/tumor characteristics and lymph node status, and to explore the use of these factors, which are all known prior to axillary dissection, in predicting lymph node status. The PCE data set contained 18,025 breast carcinoma cases diagnosed in 1990 after exclusion of women older than 79 years or with fewer than 6 lymph nodes examined. RESULTS In a multivariate logistic regression model, larger tumor size, young age, African American or Hispanic race, outer half tumor location, poor or moderate differentiation, aneuploidy, and infiltrating ductal histology were independently associated with a higher likelihood of one or more positive lymph nodes. Contrary to expectation, cases negative for estrogen receptor (ER) and progesterone receptor (PR) had a lower risk of positive lymph nodes when adjusted for other factors (odds ratio = 0.82; 95% confidence interval: 0.74–0.91) compared with cases positive for both receptors. This model accurately predicted lymph node status in 2 validation data sets (a 50% random sample of 1990 PCE data and 1992 data from the National Cancer Data Base), but was less accurate in a third, older data set (1983 PCE data). However, the percentage of cases (1990 validation set) with predicted probabilities less than 0.05 or greater than 0.95 were only 4.6% and

BACKGROUND Although growth factors such as epidermal growth factor (EGF), transforming growth factor (TGF)-α, and TGF-β are important regulators of prostate cell growth in vitro and in animal models, evidence to support their role in human prostate cancer development remains sparse. We previously showed that men without prostate cancer have concentrations of EGF and TGF-α in expressed prostatic fluid (EPF) that are individually distinct and stable over time. This study addressed whether growth factor levels in EPF are associated with the presence or progression of prostate cancer. METHODS We measured levels of immunoreactive EGF, TGF-α, and TGF-β1 in stored EPF samples from three age-matched groups: 19 men with untreated, histologically diagnosed prostate cancer (CaP), 38 with benign prostate hyperplasia (BPH), and 19 with normal prostate glands (NPD). RESULTS Median TGF-α was lower in the BPH group (0.45 ng/ml) than in either CaP (0.63 ng/ml) or NPD (0.58 ng/ml) groups (P = 0.03 and 0.12, respectively). For EGF, the median was lowest in the CaP group and highest in the NPD group (92.5 ng/ml vs. 175.5 ng/ml, P = 0.006). For TGF-β1, the median level in CaP was 2.7 times higher than the median level among all controls (6.65 ng/ml vs. 2.46 ng/ml, P = 0.002). Growth factor levels were not associated with tumor stage or Gleason score. However, the single case with distant metastases had TGF-β1 levels 23-fold higher than the CaP median. CONCLUSIONS The results suggest that at the time of CaP diagnosis, EGF levels in EPF are significantly lower, and TGF-β1 levels significantly higher, than normal. Marked overexpression of TGF-β1 in advanced CaP might be reflected in extremely high EPF levels. Prostate 40:248–255, 1999. © 1999 Wiley-Liss, Inc.

BACKGROUND CAG repeat length in exon 1 of the androgen receptor (AR) gene correlates inversely with transcriptional transactivation activity of the AR. Men with shorter AR CAG repeat lengths are at higher risk of prostate cancer. Because benign prostatic hyperplasia (BPH) is an androgen-dependent condition, we examined the hypothesis that a shorter AR gene CAG repeat length increases the risk of developing of BPH. METHODS Among 14,916 men of the Physicians' Health Study who had provided a blood sample in 1982, we measured AR gene CAG repeat lengths for 310 men who had surgery for BPH up to 7.5 years of follow-up and 1,041 controls. RESULTS Risk of surgery for BPH increased linearly with decreasing AR CAG repeat length (P (trend) = 0.03). Relative to men with a CAG repeat length ⩾25, men with a repeat length ⩽19 had an odds ratio of BPH surgery of 1.76 (95% confidence interval, 1.16–2.65). CONCLUSIONS Variability in the AR gene CAG repeat influences the development of symptomatic BPH. Prostate 39:130–134, 1999. © 1999 Wiley-Liss, Inc.

PURPOSE: To explore agreement on cancer occurrence and site among Medicare Part A, Massachusetts Cancer Registry, and death certificates. METHODS: We linked these data sources with the cohort of the population-based East Boston Senior Health Project, a component of the National Institute on Aging’s Established Populations for Epidemiologic Studies of the Elderly. The cohort consists of 905 subjects dying between January 1986 and December 1990. RESULTS: We detected the following agreements on cancer occurrence: hospitalization data and death certificates (kappa = 0.70), hospitalization and cancer registry data (kappa = 0.59), and cancer registry and death certificate data (kappa = 0.50). Measures of agreement changed little when the analyses were stratified by age, sex, calendar year and place of death, autopsy performance, cigarette smoking or alcohol consumption. Site-specific agreements were higher for colorectal and respiratory tract cancer compared to breast and prostate across all three comparisons. CONCLUSIONS: The results should assist epidemiologists to better understand the strengths and limitations of these data sources.

Carotenoids in tomatoes are hypothesized to play a chemopreventive role in prostate cancer ([1][1]); lycopene, in particular, has received considerable attention as a potent antioxidant because it seems to be the most effective quencher of singlet oxygen and free radicals ([2][2]). In addition

BACKGROUND Prostatic fluid (PF) provides a unique medium for noninvasive evaluation of critical growth and differentiation signals in the prostatic microenvironment. The purpose of this study was to establish the feasibility of measuring two prostatic mitogens, epidermal growth factor (EGF) and transforming growth factor-alpha (TGF-alpha) in PF, and specifically to quantify extraneous variability attributable to the assay itself, sample handling, or biological variation within an individual over time. METHODS PF was collected by transrectal massage from consecutive patients attending a urology clinic. Pooled PF and individual samples from 25 men with stable benign prostatic hyperplasia (BPH) were analyzed for EGF and TGF-alpha by radioimmunoassay and for total protein. RESULTS Reproducibility was adequate at dilutions as low as 1:50 (2-microliter pooled sample) and 1:5 (20 microliters) for EGF and TGF-alpha, respectively. Results were not affected by freeze-thaw cycles, time in storage, or protease inhibition in fresh PF. EGF and TGF-alpha were detectable in 100% and 92% of individual men, with respective means of 152 and 0.2 ng/ml. Correlations between two samples obtained from the same man within 12 months were highly significant (EGF r = 0.89, TGF-alpha r = 0.71). Protein concentrations were consistent over time; expression of either peptide per weight of protein rather than per volume did not improve within-man correlation. Between-man variability far exceeded within-man variability for both peptides, and was estimated to account for 84% and 61% of the total variability in EGF and TGF-alpha, respectively. There was no correlation between EGF and TGF-alpha in the same samples. CONCLUSIONS We conclude that men with BPH secrete consistent and distinct levels of EGF-related peptides in PF, and that these levels can be detected with acceptable sensitivity and precision by radioimmunoassay (RIA). Measurement of TGF-alpha, which has not been reported previously, requires a relatively larger sample.

Background Our objective was to develop and validate a multi-feature nuclear score based on image analysis of direct DNA staining, and to test its association with field effects and subsequent detection of prostate cancer (PCa) in benign biopsies. Methods Tissue sections from 39 prostatectomies were Feulgen-stained and digitally scanned (400×), providing maps of DNA content per pixel. PCa and benign epithelial nuclei were randomly selected for measurement of 52 basic morphometric features. Logistic regression models discriminating benign from PCa nuclei, and benign from malignant nuclear populations, were built and cross-validated by AUC analysis. Nuclear populations were randomly collected 5 mm from cancer foci, and from cancer-free prostates, HGPIN, and PCa Gleason grade 3–5. Nuclei also were collected from negative biopsy subjects who had a subsequent diagnosis of PCa and age-matched cancer-free controls (20 pairs). Results A multi-feature nuclear score discriminated cancer from benign cell populations with AUCs of 0.91 and 0.79, respectively, in training and validation sets of patients. In prostatectomy samples, both nuclear- and population-level models revealed cancer-like features in benign nuclei adjacent to PCa, compared to nuclei that were more distant or from PCa-free glands. In negative biopsies, a validated model with 5 variance features yielded significantly higher scores in cases than controls (P = 0.026). Conclusions A multifeature nuclear morphometric score, obtained by automated digital analysis, was validated for discrimination of benign from cancer nuclei. This score demonstrated field effects in benign epithelial nuclei at varying distance from PCa lesions, and was associated with subsequent PCa detection in negative biopsies. Impact This nuclear score shows promise as a risk predictor among men with negative biopsies and as an intermediate biomarker in Phase II chemoprevention trials. The results also suggest that subvisual disturbances in nuclear structure precede the development of pre-neoplastic lesions.

BACKGROUND It has been postulated that different biologic characteristics of a tumor might account for at least a portion of the disparity in breast cancer survival for patients across racial and ethnic groups. The hormone receptor status of breast tumors is one characteristic with prognostic significance. Results of several studies indicate a higher proportion of estrogen receptor negative (ER-) breast tumors among black or Hispanic women compared with non-Hispanic whites. We investigated whether the association between race and ethnicity and the joint combination of ER and progesterone receptor (PR) status was independent of age and other tumor characteristics at diagnosis. METHODS Relevant data were obtained from 13,239 breast cancer cases identified as part of the 1990 Patient Care Evaluation Study of Breast Cancer. RESULTS In univariate analysis, the proportions of ER+PR+, ER+PR−, ER−PR+, and ER−PR− tumors among non-Hispanic whites were 59%, 15%, 6%, and 20%, respectively; among Hispanics the proportions were 58%, 12%, 8%, and 22%, respectively; and among blacks the proportions were 44%, 14%, 7%, and 35%, respectively. After controlling for age, tumor size, and histology, using polychotomous logistic regression, there was no difference in hormone-receptor status between Hispanic and non-Hispanic white women. However, ER−PR− tumors were more likely to occur in blacks than in non-Hispanic whites (odds ratio = 1.8, 95% confidence interval = 1.6, 2.1) CONCLUSIONS The results of this study corroborate differences in hormone-receptor status between non-Hispanic white and black women, but not between Hispanics. Future studies of breast cancer should examine differences in epidemiologic risk factors between blacks and whites stratified on ER/PR status. Cancer 1996; 77:1465-71.

We assessed the relation of plasma hormone levels and nonhormonal factors with subsequent occurrence of surgical treatment for benign prostatic hyperplasia (BPH) among participants in the Physicians' Health Study. Frozen plasma samples, collected at the study onset, were available for 320 men who developed surgically treated BPH up to 9 years later and for 320 age-matched controls. Plasma testosterone (T), dihydrotestosterone (DHT), androstenedione, estradiol (E2), and estrone (E1) were measured for each case-control pair. In unadjusted analyses, none of the hormones or hormone ratios were associated with BPH; for example, for T and E2 the odds ratios (OR) comparing the highest quintile (Q5) with the lowest (Q1) were 0.74 (95% CI = 0.42, 1.30) and 1.07 (95% CI = 0.51, 2.22), respectively. However, in multivariate analyses controlling diastolic blood pressure, exercise, alcohol, E1, and DHT:T ratio, we observed a strong trend for increasing risk across quintiles for E2 (Q5 vs. Q1 OR = 3.56, P trend = 0.009), and a weak inverse trend for E1 (Q5 vs Q1 OR = 0.51, P trend = 0.07). The excess risk associated with E2 was confined to men with relatively low androgen levels. Three nonhormonal factors previously suspected as risk factors were independently associated with surgical BPH in these data. The OR for a 1-mm Hg difference in diastolic blood pressure was 1.04 (95% CI = 1.01, 1.07). Alcohol use and infrequent exercise were inversely associated with risk of BPH surgery; however, risk estimates were not consistent across categories of exercise and alcohol frequency. Our results indicate that normal variation in circulating androgen levels does not influence development of BPH, but that variation in estrogen levels might be important.

We provide current information on the use of PSA testing for the evaluation of men at risk for prostate cancer, and the risks and benefits of early detection.The report is a summary of the American Urological Association PSA Best Practice Policy 2009. The summary statement is based on a review of the current professional literature, clinical experience and the expert opinions of a multispecialty panel. It is intended to serve as a resource for physicians, other health care professionals, and patients. It does not establish a fixed set of guidelines, define the legal standard of care or pre-empt physician judgment in individual cases.There are two notable differences in the current policy. First, the age for obtaining a baseline PSA has been lowered to 40 years. Secondly, the current policy no longer recommends a single, threshold value of PSA, which should prompt prostate biopsy. Rather, the decision to proceed to prostate biopsy should be based primarily on PSA and DRE results, but should take into account multiple factors including free and total PSA, patient age, PSA velocity, PSA density, family history, ethnicity, prior biopsy history and comorbidities.Although recently published trials show different results regarding the impact of prostate cancer screening on mortality, both suggest that prostate cancer screening leads to overdetection and overtreatment of some patients. Therefore, men should be informed of the risks and benefits of prostate cancer screening before biopsy and the option of active surveillance in lieu of immediate treatment for certain men diagnosed with prostate cancer.

Sex hormones play an important role in the growth and development of the prostate, and low androgen levels have been suggested to carry an adverse prognosis for men with prostate cancer (PCa).To examine the association between prediagnostic circulating sex hormones and lethal PCa in two prospective cohort studies, the Physicians' Health Study (PHS) and the Health Professionals Follow-up Study (HPFS).We included 963 PCa cases (700 HPFS; 263 PHS) that provided prediagnostic blood samples, in 1982 for PHS and in 1993-1995 for HPFS, in which circulating sex hormone levels were assayed.The primary end point was lethal PCa (defined as cancer-specific mortality or development of metastases), and we also assessed total mortality through March 2011. We used Cox proportional hazards models to evaluate the association of prediagnostic sex hormone levels with time from diagnosis to development of lethal PCa or total mortality.PCa cases were followed for a mean of 12.0±4.9 yr after diagnosis. We confirmed 148 cases of lethal PCa and 421 deaths overall. Using Cox proportional hazard models, we found no significant association between quartile of total testosterone, sex hormone binding globulin (SHBG), SHBG-adjusted testosterone, free testosterone, dihydrotestosterone, androstanediol glucuronide, or estradiol and lethal PCa or total mortality. In subset analyses stratified by Gleason score, TNM stage, age, and interval between blood draw and diagnosis, there was also no consistent association between lethal PCa and sex hormone quartile.We found no overall association between prediagnostic circulating sex hormones and lethal PCa or total mortality. Our null results suggest that reverse causation may be responsible in prior studies that noted adverse outcomes for men with low circulating androgens.

Phytanic acid is a saturated fatty acid found predominantly in red meat and dairy products and may contribute to increases in prostate cancer risk that are observed with higher intakes of these foods. We constructed a novel summary measure of phytanic acid intake and prospectively examined its association with prostate cancer risk in the Alpha-Tocopherol, Beta-Carotene Cancer Prevention Study--a cohort of Finnish male smokers aged 50-69 years. Diet was assessed at baseline in 27,111 participants using a validated 276-item dietary questionnaire. Since phytanic acid is not currently included in food composition tables, we used the published phytanic acid content of 151 major food items to estimate total daily intake. During up to 21 years of follow-up, a total of 1,929 incident prostate cancer cases (including 438 advanced cases) were identified. Higher phytanic acid intake, though unrelated to the risk of localized disease [relative risks (RR) and 95% confidence intervals (CI) for increasing quartiles of intake = 1.00 (ref), 0.83 (0.68-1.01), 0.76 (0.62-0.94) and 0.91 (0.74-1.13); p trend = 0.23], was associated with increased risks of advanced prostate cancer [RR and 95% CI = 1.00 (ref), 1.43 (1.09-1.89), 1.31 (0.99-1.75) and 1.38 (1.02-1.89); p trend = 0.06]. This association appeared to be driven predominantly by phytanic acid obtained from dairy products (particularly butter). Our study indicates that phytanic acid may contribute to previously observed associations between high-fat animal foods (particularly dairy products) and prostate cancer risk, although some caution is warranted as it may be acting as a surrogate marker of dairy fat.

Functional polymorphisms in endogenous antioxidant defense genes including manganese superoxide dismutase (MnSOD), catalase (CAT), and glutathione peroxidase (GPX-1) have been linked with risk of cancer at multiple sites. Although it is presumed that these germline variants impact disease risk by altering the host's ability to detoxify mutagenic reactive oxygen species, very few studies have directly examined this hypothesis. Concentrations of 8-isoprostane F2α (8-iso-PGF2α) and 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxoxdG)-sensitive indicators of lipid peroxidation and DNA oxidation, respectively-were measured in 24-h urine samples obtained from 93 healthy, premenopausal women participating in a dietary intervention trial. In addition, DNA was extracted from blood for genotyping of MnSOD Val16Ala, CAT-262 C T, and GPX1 Pro198Leu genotypes by Taqman assay. Although geometric mean concentrations of 8-iso-PGF2(α) and 8-oxoxdG varied across several study characteristics including race, education level, body mass index, and serum antioxidant levels, there was little evidence that these biomarkers differed across any of the examined genotypes. In summary, functional polymorphisms in endogenous antioxidant defense genes do not appear to be strongly associated with systemic oxidative stress levels in young, healthy women.

BACKGROUND: Although some evidence suggests that dietary fat intake is related to prostate cancer, epidemiologic studies have been inconsistent. PURPOSE: Our purpose was to assess the association between plasma lipid levels, particularly linoleic and alpha-linolenic acids, and the development of prostate cancer. METHODS: In 1982, at the start of the Physicians' Health Study, 14916 U.S. male physicians provided plasma samples, which were frozen at -82 degrees C. Data accumulated from a series of questionnaires were used to assess the intake of various foods. We used a nested case-control design to compare the fatty acid compositions in plasma from 120 men who later developed prostate cancer with 120 matched controls who did not. Individual fatty acids were measured in plasma as a percentage of total fatty acids, using capillary gas chromatography. Conditional logistic regression models were used to obtain odds ratio estimates while adjusting simultaneously for the effects of one or more potential confounders. RESULTS: The relative risks (RRs) of prostate cancer for men in successively higher quartiles of plasma alpha-linolenic acid level were 3.0 (95% confidence interval [CI] = 1.2-7.3), 3.4 (95% CI = 1.6-7.5), and 2.1 (95% CI = 0.9-4.9), compared with those with levels below the detection threshold (P trend =.03). For linoleic acid, RRs in successively higher quartiles were 0.7 (95% CI = 0.4-1.5), 0.8 (95% CI = 0.4-1.6), and 0.6 (95% CI = 0.3-1.3), with the lowest quartile as referent (P trend =.24). The effect estimates were not notably altered by adjustment for exercise, body mass, meat and dairy consumption, or other fatty acid levels in the plasma. The RR for eating red meat at least five times per week compared with less than once a week was 2.5 (95% CI = 0.9-6.7) and was little changed by adjustment for alpha-linolenic acid, although alpha-linolenic acid levels were correlated with intake of red meat and butter. The association of alpha-linolenic acid levels with prostate cancer was greater among men with low linoleic acid and reduced meat intake. CONCLUSIONS: These results suggest that low plasma levels of alpha-linolenic acid might be associated with reduced risk of prostate cancer, independently of high meat intake. High linoleic acid and low marine fatty oils were not associated with increased risk, as previously hypothesized. IMPLICATIONS: The effects of dietary alpha-linolenic acid, particularly from vegetable sources, warrant further study. The effects of dietary linoleic acid and marine fatty acids seen in animal bioassays might not apply to human prostate cancer.

Summary Previous investigations have linked decreased nuclear expression of the cell cycle inhibitor p27 with poor outcome in prostate cancer. However, these reports are inconsistent regarding the magnitude of that association and its independence from other predictors. Moreover, cytoplasmic translocation of p27 has been proposed as a negative prognostic sign. Given the cost and accuracy limitations of manual scoring, particularly of tissue microarrays, we determined if laser-based fluorescence microscopy could provide automated analysis of p27 in both nuclear and cytoplasmic locations and, thus, clarify its significance as a prognostic biomarker. We constructed tissue microarrays covering 202 recurrent cases (rising prostate-specific antigen) and 202 matched controls without recurrence. Quadruplicate tumor samples encompassed 5 slides and 1616 cancer histospots. Cases and controls matched on age, Gleason grade, stage, and hospital. We immunolabeled epithelial cytoplasm with Alexafluor 647, p27 with Alexafluor 488, and nuclei with 4c6-diamidino-2-phenylindole·2HCl. Slides were scanned on an iCys laser scanning cytometer (CompuCyte Corp, Cambridge, MA). Nuclear crowding required a stereological approach—random arrays of circles (phantoms) were layered on images and the content of each phantom was analyzed in scatter plots. Both nuclear and cytoplasmic p27 were significantly lower in cases versus controls ( P = .014 and P = .004, respectively). Regression models controlling for matching variables plus prostate-specific antigen showed strong linear trends for increased risk of recurrence with lower p27 in both nucleus and cytoplasm (highest versus lowest quartile; odds ratio, 0.35; P = .006). Manual scoring identified an inverse association between p27 expression and tumor grade but no independent association with recurrence. In conclusion, we developed an automated method for subcellular scoring of p27 without the need to segment individual cells. Our method identified a strong relationship, independent of tumor grade, stage, and prostate-specific antigen, between p27 expression—regardless of subcellular location—and prostate cancer recurrence. This relationship was not observed with manual scoring.

A potent bladder carcinogen for workers in the dye industry, 4-aminobiphenyl (4-ABP), is present in environmental tobacco smoke and has been shown to bond covalently with hemoglobin.The goal of this study was to examine the relationship between exposure to environmental tobacco smoke and levels of 4-ABP-hemoglobin adducts in nonsmoking pregnant women and to compare adduct levels in those women with levels in smoking pregnant women.A questionnaire on smoking and exposure to environmental tobacco smoke was administered to 15 pregnant women who smoked cigarettes and 40 who did not smoke. Exposure was quantified for 1 week with a personal diary and by air sampling with a monitor worn by each woman. The monitor collected nicotine by passive diffusion to a filter treated with sodium bisulfate, and the deposit on the filter was analyzed by gas chromatography. Aliquots of maternal blood and cord blood collected during delivery were analyzed for 4-ABP-hemoglobin adducts by gas chromatography with negative ion chemical ionization mass spectrometry.The mean adduct level in smokers (184 pg of 4-ABP per gram of hemoglobin) was substantially higher than that in nonsmokers (22 pg/g). This difference was statistically significant. Among nonsmokers, the levels of 4-ABP adducts increased significantly with increasing environmental tobacco smoke level (P = .009). Those in the lowest exposure category (0.5 micrograms/m3 weekly average nicotine) had median 4-ABP-hemoglobin adduct levels of 15 pg of 4-ABP per gram of hemoglobin, while those in the highest exposure category (or = 2.0 micrograms/m3) had median levels of 26 pg/g. Nonsmokers in this study had a median adduct level of 20 pg/g, and smokers had a median level of 143 pg/g.4-ABP-hemoglobin adduct levels in nonsmokers were 14% of the levels in smokers, which is consistent with findings of 20% in two other studies. Nonsmokers may receive a nontrivial dose of carcinogens from environmental tobacco smoke proportional to their exposure to environmental tobacco smoke.The relationship between environmental tobacco smoke exposure and 4-ABP-hemoglobin adduct levels supports epidemiologic evidence that environmental tobacco smoke is carcinogenic to passive smokers.

Purpose The European Randomized Study of Screening for Prostate Cancer (ERSPC) reported a 20% mortality reduction with prostate-specific antigen (PSA) screening. However, they estimated a number needed to screen (NNS) of 1,410 and a number needed to treat (NNT) of 48 to prevent one prostate cancer death at 9 years. Although NNS and NNT are useful statistics to assess the benefits and harms of an intervention, in a survival study setting such as the ERSPC, NNS and NNT are time specific, and reporting values at one time point may lead to misinterpretation of results. Our objective was to re-examine the effect of varying follow-up times on NNS and NNT using data extrapolated from the ERSPC report. Materials and Methods On the basis of published ERSPC data, we modeled the cumulative hazard function using a piecewise exponential model, assuming a constant hazard of 0.0002 for the screening and control groups for years 1 to 7 of the trial and different constant rates of 0.00062 and 0.00102 for the screening and control groups, respectively, for years 8 to 12. Annualized cancer detection and drop-out rates were also approximated based on the observed number of individuals at risk in published ERSPC data. Results According to our model, the NNS and NNT at 9 years were 1,254 and 43, respectively. Subsequently, NNS decreased from 837 at year 10 to 503 at year 12, and NNT decreased from 29 to 18. Conclusion Despite the seemingly simplistic nature of estimating NNT, there is widespread misunderstanding of its pitfalls. With additional follow-up in the ERSPC, if the mortality difference continues to grow, the NNT to save a life with PSA screening will decrease.

Epidemiologic studies have established that pregnancy has a bidirectional, time-dependent effect on breast cancer risk; a period of elevated risk is followed by a long-term period of protection. The purpose of the present study was to determine whether pregnancy and involution are associated with gene expression changes in the normal breast, and whether such changes are transient or persistent. We examined the expression of a customized gene set in normal breast tissue from nulliparous, recently pregnant (0-2 years since pregnancy), and distantly pregnant (5-10 years since pregnancy) age-matched premenopausal women. This gene set included breast cancer biomarkers and genes related to immune/inflammation, extracellular matrix remodeling, angiogenesis, and hormone signaling. Laser capture microdissection and RNA extraction were done from formalin-fixed paraffin-embedded reduction mammoplasty and benign biopsy specimens and analyzed using real-time PCR arrays containing 59 pathway-specific and 5 housekeeping genes. We report 14 of 64 (22%) of the selected gene set to be differentially regulated (at P < 0.05 level) in nulliparous versus parous breast tissues. Based on gene set analysis, inflammation-associated genes were significantly upregulated as a group in both parous groups compared with nulliparous women (P = 0.03). Moreover, parous subjects had significantly reduced expression of estrogen receptor α (ERα, ESR1), progesterone receptor (PGR), and ERBB2 (Her2/neu) and 2-fold higher estrogen receptor-β (ESR2) expression compared with nulliparous subjects. These initial data, among the first on gene expression in samples of normal human breast, provide intriguing clues about the mechanisms behind the time-dependent effects of pregnancy on breast cancer risk. Cancer Prev Res; 3(3); 301–11

Purpose To introduce a novel approach for the time-dependent quantification of risk factors for prostate cancer (PCa) detection after an initial negative biopsy. Patients and Methods Data for 1,871 men with initial negative biopsies and at least one follow-up biopsy were available. Piecewise exponential regression models were developed to quantify hazard ratios (HRs) and define cumulative incidence curves for PCa detection for subgroups with specific patterns of risk factors over time. Factors evaluated included age, race, serum prostate-specific antigen (PSA) concentration, PSA slope, digital rectal examination, dysplastic glands or prostatitis on biopsy, ultrasound gland volume, urinary symptoms, and number of negative biopsies. Results Four hundred sixty-five men had PCa detected, after a mean follow-up time of 2.8 years. All of the factors were independent predictors of PCa detection except for PSA slope, as a result of its correlation with time-dependent PSA level, and race. PSA (HR = 3.90 for > 10 v 2.5 to 3.9 ng/mL), high-grade prostatic intraepithelial neoplasia/atypical glands (HR = 2.97), gland volume (HR = 0.39 for > 50 v < 25 mL), and number of repeat biopsies (HR = 0.36 for two v zero repeat biopsies) were the strongest predictors. Men with high-risk versus low-risk event histories had a 20-fold difference in PCa detection over 5 years. Conclusion Piecewise exponential models provide an approach to longitudinal analysis of PCa risk that allows clinicians to see the interplay of risk factors as they unfold over time for individual patients. With these models, it is possible to identify distinct subpopulations with dramatically different needs for monitoring and repeat biopsy.

Field cancerization, which is not yet well-characterized in the prostate, occurs when large areas of an organ or tissue surface are affected by a carcinogenic insult, resulting in the development of multi-focal independent premalignant foci and molecular lesions that precede histological change.Herein, we review the cumulative body of evidence concerning field effects in the prostate and critically evaluate the methods available for the identification and validation of field effect biomarkers. Validated biomarkers for field effects have an important role to play as surrogate endpoint biomarkers in Phase II prevention trials and as clinical predictors of cancer in men with negative biopsies.Thus far, field effects have been identified involving nuclear morphometric changes, gene expression, protein expression, gene promoter methylation, DNA damage and angiogenesis. In addition to comparing cancer-adjacent benign tissue to more distant areas or to "supernormal" tissue from cancer-free organs, investigators can use a nested case-control design for negative biopsies that offers a number of unique advantages.True carcinogenic field effects should be distinguished from secondary responses of the microenvironment to a developing tumor, although the latter may still lead to useful clinical prediction tools. Prostate 69: 1470-1479, 2009. (c) 2009 Wiley-Liss, Inc.

We evaluated the beliefs of second-year medical students regarding critical issues in the AIDS epidemic before and after an intensive two-day training symposium. Paired responses, collected for three consecutive years, were available for 187 students. Our results indicate that, although the students generally held progressive beliefs on prevention and public policy prior to training, a significant minority held views that differ from expert opinion. After the training experience, we observed substantial shifts of opinion on several issues. For example, among those who initially believed that physicians should record results of HIV testing in patient charts without patient permission, 45% shifted to disagreement. On mandatory premarital testing, 43% of those initially supporting it shifted to disagreement. On some issues, such as contact tracing, where expert opinion is less clear, student opinion moved very little overall. Over the three-year period, we found evidence of a trend towards less concern over HIV transmission to men from causal sex with infected female partners and a trend towards acceptance of glove-wearing when handling all blood specimens. In conclusion, we demonstrate that the beliefs of medical students concerning difficult issues posed by the AIDS epidemic can be changed substantially in the short-term by a specially designed intensive educational program. Reinforcement and persistence of these changes should now be a concern for medical educators.

BACKGROUND: Some, but not all, published results have shown an association between circulating blood levels of some insulin-like growth factors (IGFs) and their binding proteins (IGFBPs) and the subsequent risk for prostate cancer. PURPOSE: To assess the association between levels of IGFs and IGFBPs and the subsequent risk for prostate cancer. DATA SOURCES: Studies identified in PubMed, Web of Science, and CancerLit. STUDY SELECTION: The principal investigators of all studies that published data on circulating concentrations of sex steroids, IGFs, or IGFBPs and prostate cancer risk using prospectively collected blood samples were invited to collaborate. DATA EXTRACTION: Investigators provided individual participant data on circulating concentrations of IGF-I, IGF-II, IGFBP-II, and IGFBP-III and participant characteristics to a central data set in Oxford, United Kingdom. DATA SYNTHESIS: The study included data on 3700 men with prostate cancer and 5200 control participants. On average, case patients were 61.5 years of age at blood collection and received a diagnosis of prostate cancer 5 years after blood collection. The greater the serum IGF-I concentration, the greater the subsequent risk for prostate cancer (odds ratio [OR] in the highest vs. lowest quintile, 1.38 [95% CI, 1.19 to 1.60]; P < 0.001 for trend). Neither IGF-II nor IGFBP-II concentrations were associated with prostate cancer risk, but statistical power was limited. Insulin-like growth factor I and IGFBP-III were correlated (r = 0.58), and although IGFBP-III concentration seemed to be associated with prostate cancer risk, this was secondary to its association with IGF-I levels. Insulin-like growth factor I concentrations seemed to be more positively associated with low-grade than high-grade disease; otherwise, the association between IGFs and IGFBPs and prostate cancer risk had no statistically significant heterogeneity related to stage or grade of disease, time between blood collection and diagnosis, age and year of diagnosis, prostate-specific antigen level at recruitment, body mass index, smoking, or alcohol intake. LIMITATIONS: Insulin-like growth factor concentrations were measured in only 1 sample for each participant, and the laboratory methods to measure IGFs differed in each study. Not all patients had disease stage or grade information, and the diagnosis of prostate cancer may differ among the studies. CONCLUSION: High circulating IGF-I concentrations are associated with a moderately increased risk for prostate cancer.

Contains fulltext : 51583.pdf (Publisher’s version ) (Closed access) We examined the hypothesis that serum concentrations of circulating androgens and sex hormone binding globulin (SHBG) are associated with risk for prostate cancer in a case-control study nested in the European Prospective Investigation into Cancer and Nutrition (EPIC). Concentrations of androstenedione, testosterone, androstanediol glucuronide and SHBG were measured in serum samples for 643 prostate cancer cases and 643 matched control participants, and concentrations of free testosterone were calculated. Conditional logistic regression models were used to calculate odds ratios for risk of prostate cancer in relation to the serum concentration of each hormone. After adjustment for potential confounders, there was no significant association with overall risk for prostate cancer for serum total or free testosterone concentrations (highest versus the lowest thirds: OR, 1.02; 95% CI, 0.73-1.41 and OR, 1.07, 95% CI, 0.74-1.55, respectively) or for other androgens or SHBG. Subgroup analyses showed significant heterogeneity for androstenedione by cancer stage, with a significant inverse association of androstenedione concentration and risk for advanced prostate cancer. There were also weak positive associations between free testosterone concentration and risk for total prostate cancer among younger men and risk for high-grade disease. In summary, in this large nested case-control study, concentrations of circulating androgens or SHBG were not strongly associated with risk for total prostate cancer. However, our findings are compatible with a positive association of free testosterone with risk in younger men and possible heterogeneity in the association with androstenedione concentration by stage of disease; these findings warrant further investigation.

Background: The nucleo-cytoplasmic shuttle protein nucleophosmin (NPM1) is mutated in 50% of cytogenetically normal AML cases (CN-AML) resulting in the nuclear export of this protein and a favorable prognosis. The mechanism by which mutated NPM1 (NPM1mut) confers this advantage in CN-AML is unclear. We have previously demonstrated that FOXM1, an oncogenic transcription factor, co-localizes with NPM1 in cancer cells and NPM-binding is required for sustaining the level of FOXM1. In the NPM1mut AML cell line OCI-AML3, as detected by immunofluorescence, NPM1 and FOXM1 co-localize in the cytoplasm where FOXM1 is transcriptionally inactive. We hypothesize that improved outcomes in the subset of NPM1mut AML is partly attributable to the cytoplasmic relocalization and functional inactivation of FOXM1. Methods: Patient samples were identified retrospectively using the University of Illinois Cancer Center Registry (IRB #: 2013-1245). Clinical, laboratory and molecular data were collected by chart review. Formalin fixed paraffin embedded bone marrow biopsies were retrieved to prepare slides. Patients with favorable or unfavorable karyotypes were excluded and only biopsies with >70% blasts were included. We performed a 2-target staining protocol with fluorescent antibodies to visualize NPM1 and FOXM1 in the same cells using the Vectra multispectral tissue imaging system. This system combines machine learning capability (inForm® software) with acquisition of spectral data from each pixel in the image, so that multiple targets can be quantified in the same image. Leukemia cell lines with stable knockdown of FOXM1 were generated using lentiviral transduction of HL60 cells (NPM1wt) transduced with PLKO.1-shFOXM1 or PLKO.1-empty vector. In parallel, OCI-AML3 cells (NPM1mut) were transduced with pBabe control or pBabe-FOXM1 expressing retrovirus to induce overexpression of FOXM1. Results: FOXM1 is a well-characterized oncogenic transcription factor. In this study we examined its role in AML. In a series of AML cell lines, cellular fractionation followed by immunoblotting revealed strong nuclear expression of FOXM1 in HL-60 and KG1, both of which express the wild-type NPM1 gene. In comparison the OCI-AML3 cell line, carrying the mutated NPM1 gene, showed predominantly cytoplasmic expression of FOXM1 where it is inactive. To better understand the role of FOXM1 in mediating chemoresistance in AML, we inhibited FOXM1 expression in the HL60 leukemia cell line. Stable knockdown of FOXM1 in this cell line resulted in increased sensitivity to the chemotherapeutic agent cytarabine compared to empty vector transduced cells. This was demonstrated by increased caspase-3 cleavage as detected by immunoblotting and decreased cell viability by Trypan blue exclusion after 24 hour exposure to the drug. We then proceeded to validate our observations of the FOXM1-NPM interaction in primary AML samples. Using quantitative data generated from the Vectra imaging of 20 AML pre-treatment bone marrow biopsies we show a strong correlation between the nuclear: cytoplasmic ratios of FOXM1 and NPM, Pearson correlation co-efficient r=0.87, which is indicative of cellular co-localization. When the patients are stratified in 2 groups based on NPM1 mutation status, mean cytoplasmic expression of FOXM1 is 2-fold higher in NPM1mut patients (36% versus 18%). Conclusion: We demonstrate that FOXM1, an oncogenic transcription factor is predominantly cytoplasmic and thereby inactive when NPM1 is mutated in AML. We show the functional significance of FOXM1 in mediating chemoresistance in this disease using AML cell lines. Using a novel quantitative imaging modality we establish for the first time the cellular co-localization of FOXM1 and NPM1 in AML primary blast cells. Studies are ongoing to manipulate FOXM1 expression in additional leukemia cell lines and xenograft models of AML to confirm our findings that FOXM1 inhibition in AML leads to increased sensitivity to chemotherapy. Disclosures No relevant conflicts of interest to declare.

Mutations in BRCA1 are associated with familial as well as sporadic aggressive subtypes of breast cancer. However, it is not clear to what extent BRCA1 expression or its subcellular localization contributes to breast cancer progression. The goal of this analysis was to examine the differential expression and subcellular localization of BRCA1 among normal breast tissue and breast cancer cases, and whether it could serve as an additional prognostic marker in an ethnically diverse sample of 287 patients (86 Non-Hispanic White, 84 Hispanic and 116 African American). Tissue microarrays (TMAs) were constructed from invasive breast cancer samples obtained from the Breast Cancer Care in Chicago study in addition to 46 age and race-matched normal breast tissue samples. In these TMAs, BRCA1 was immunolabeled with Alexa647; epithelial cytoplasm with Alexa488; and nuclei with DAPI. Slides were visualized and quantified using “VECTRA Automated Multispectral Image Analysis System” and “InForm software”. BRCA1 expression was evaluated based on the percentage of positive cells and staining intensity using the H-score. The H score is a product of the percentage of cells (0-100%) in each intensity category (0, 1+, 2+ and 3+). The final score is on a continuous scale between 0 and 300. The average nuclear score for BRCA1 (Nuc) was higher than that of the cytoplasmic score (Cyto) in normal breast tissue (Nuc: 157.7 ±6.0; Cyto: 150.7±7.5, p = 0.02) and breast cancer cells (Nuc: 140.7 ±3.3; Cyto: 118.0±3.4, p Note: This abstract was not presented at the meeting. Citation Format: Abeer Mostafa Mahmoud, Umaima Al-alem, Virgilia Macias, Ryan J. Deaton, Andre Balla, Peter Gann, Garth Rauscher. BRCA1 protein expression and subcellular localization in primary breast cancer: automated digital microscopy analysis of tissue microarrays. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr LB-019. doi:10.1158/1538-7445.AM2015-LB-019

Approximately 3.2 million Americans are chronically infected with Hepatitis C, forming the largest group of persons at high risk for HCC, with an annual incidence of 3-4%. Increased oxidative stress is regarded as the major mechanism of progression in chronic liver disease (CLD) and subsequent HCC. Hepatic iron overload facilitates the production of reactive oxygen species (ROS), thereby inducing liver damage and fibrosis. We have previously shown that retinoid and carotenoid depletion occurs early in disease and may represent a major modifiable risk. This study aims to determine, in CLD patients and controls, whether retinoid levels modify the association between serum iron concentrations and: (a) oxidative stress biomarkers in urine (b) tissue changes associated with hepatic stellate cell (HSC) activation, and (c) CLD progression. Patients with HCV undergoing liver biopsy (n = 69) provided fasting blood, fresh tissue, urine and completed a diet history questionnaire (DHQ). Serum and questionnaire data from healthy volunteers (n = 11), normal liver tissue from public repositories and patients without liver disease (n = 11) were also collected. Urinary isoprostanes, serum and tissue retinoid concentrations were obtained by LC-MS-MS. Serum iron, ferritin and transferrin were quantitated. Immunohistochemistry for αSMA was performed on FFPE sections and subsequently quantified via digital image analysis. Associations between urinary isoprostanes, αSMA levels, serum iron, serum and tissue retinoids were assessed using Spearman correlation coefficients and non-parametric tests were utilized to test differences among disease severity groups. There was a significant increasing trend for serum transferrin levels with progressive fibrosis, but not for serum iron and ferritin concentrations. All subjects had transferrin saturation within the normal range. Serum β-carotene and lycopene were inversely associated with serum ferritin concentrations (r = -0.29, P < 0.01; r = -0.29, P < 0.01) however, this relationship was not observed for other serum iron measurements. Serum iron measurements were not associated with hepatic retinoids/carotenoids, αSMA expression or urinary isoprostanes. Tissue retinyl palmitate was inversely and significantly correlated with hepatic αSMA expression (r = -0.3, P < 0.02). Serum iron overload was not evident in this population even though a depletion of serum retinoids/carotenoids is observed. The inverse correlation observed between serum ferritin and retinoids/carotenoids suggests a possible joint effect in CLD patients. Our data provide confirmation that hepatic retinoid loss is accompanied by HSC activation, a key player in fibrosis. Further evaluation of hepatic iron status is warranted to assess the relationships between iron and retinoids in HCV. Citation Format: Yachana Kataria, Ryan Deaton, Erika Enk, Milita Petrauskaite, Scott Cotler, Donald Jensen, Peter Gann. Joint relationship between iron and retinoids in patients at high-risk for liver cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2821. doi:10.1158/1538-7445.AM2015-2821

Background: Conventional methods for predicting prostate cancer (PCa) aggressiveness, which rely heavily on the Gleason score of architectural pattern, remain imperfect. We have developed an approach that combines machine vision and machine learning analysis of routine H&E stained tumors to discriminate recurrent from non-recurrent patients after prostatectomy, based on architectural, color, and nuclear patterns. Methods: We used a set of H&E-stained TMAs containing quadruplicate cores from 176 recurrent cases of PCa and 154 non-recurrent controls, frequency matched on age, Gleason, pathological stage and race. Using software specifically adaptable for computational pathology with hierarchical object recognition (Definiens Developer XD®), we processed images of each tumor at two spatial scale levels: a larger scale for recognition of tissue compartments (epithelium, stroma, lumen and inflammation), and a smaller scale for recognition of normal and atypical nuclei in each of the compartments. Magnification of 20x was used for both levels. At the tissue level, we used multi-resolution segmentation to segment homogeneous superpixels, which were then classified using machine learning into epithelium, stroma. or ambiguous tissue classified as intermediate between epithelium and stroma. We used relative darkness in a spatial neighborhood, size, and shape criteria to segment typical and atypical nuclei. Relational features among objects within and between the tissue and nuclear levels were also created. The resultant feature library contained 884 base features and 3,551 total variables. We used several stages of iterative L1 penalized logistic regression and 5-fold cross-validation to reduce variables and obtain AUC estimates. Variable stability criteria applied across the folds further minimized model overfitting. Results: PSA, Gleason and CAPRA-s achieved AUCs < 0.60 for recurrence in this matched dataset. A fifth stage L1 model containing 18 histometric features had cross-validation AUC = 0.81 (95% CI: 0.76-0.85). A more permissive 74 feature model had AUC = 0.95 (95% CI: 0.93-0.97). Subgroup analysis restricted to Gleason 7 cases produced AUCs ranging from 0.79-0.93 with varying model sparseness. Including pre-surgical PSA, Gleason grade or CAPRA-s score did not change model performance. Notably, leading features were frequently derived from stromal or ambiguously stromal regions and involved nuclear shape or texture. Their independence from tumor grading reflected subject matching and lack of explicit accounting for nuclear or stromal characteristics in the Gleason criteria. Conclusion: A computational pathology approach, applied to discrimination of recurrent and non-recurrent PCa in routine H&E stains, shows considerable promise and appears capable of adding to the predictive power of Gleason and CAPRA scoring. We will conduct additional validation studies and extend this work to biopsy samples. Citation Format: Amit Sethi, Lingdao Sha, Ryan J. Deaton, Virgilia Macias, Andrew H. Beck, Peter H. Gann. Computational pathology for predicting prostate cancer recurrence. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr LB-285. doi:10.1158/1538-7445.AM2015-LB-285

Selenium-Binding Protein 1 (SBP1, SELENBP1, hSP56) is a selenium-associated protein shown to be at lower levels in tumors, and its lower levels are frequently predictive of a poor clinical outcome. Distinguishing indolent from aggressive prostate cancer is a major challenge in disease management. Associations between SBP1 levels, tumor grade, and disease recurrence following prostatectomy were investigated by duplex immunofluorescence imaging using a tissue microarray containing tissue from 202 prostate cancer patients who experienced biochemical (PSA) recurrence after prostatectomy and 202 matched control patients whose cancer did not recur. Samples were matched by age, ethnicity, pathological stage and Gleason grade, and images were quantified using the Vectra multispectral imaging system. Fluorescent labels were targeted for SBP1 and cytokeratins 8/18 to restrict scoring to tumor cells, and cell-by-cell quantification of SBP1 in the nucleus and cytoplasm was performed. Nuclear SBP1 levels and the nuclear to cytoplasm ratio were inversely associated with tumor grade using linear regression analysis. Following classification of samples into quartiles based on the SBP1 levels among controls, tumors in the lowest quartile were more than twice as likely to recur compared to those in any other quartile. Inducible ectopic SBP1 expression reduced the ability of HCT-116 human tumor cells to grow in soft agar, a measure of transformation, without affecting proliferation. Cells expressing SBP1 also demonstrated a robust induction in the phosphorylation of the p53 tumor suppressor at serine 15. These data indicate that loss of SBP1 may play an independent contributing role in prostate cancer progression and its levels might be useful in distinguishing indolent from aggressive disease.

Not long ago, anyone applying for funds to identify risk factors for prostate cancer could elicit support for their proposal by arguing that relatively little effort had been spent thus far. While we still can claim that few, if any, risk factors can be called 'established', we've lost the element of sympathy for the assertion that prostate cancer etiology is still a neglected research area. Witness this issue of the journal and the last, which contain six papers dealing with the epidemiology of prostate cancer. We appear to be undergoing a transition in this field from a lack of interesting causal hypotheses to an almost embarrassing set of riches. The case for the existence of important dietary risk factors in prostate cancer, including those that might modify risk even at late stages of tumor development, is remarkably strong. The international variation in prostate cancer mortality exceeds even that of breast cancer, and appears to stem from differences in the rate of progression from the very small, latent lesions observed at autopsy.' Less well-appreciated, though, is the observation that migrants to Western countries from Asia can adopt the risk level of the host country within one generation, perhaps even within several years of migration.2 Even before the recent burst of information, we knew that something about a pattern of eating characterized by high animal fat intake was strongly associated with prostate cancer risk. Indeed, the international association between incidence rates and per capita fat disappearance might even be stronger for prostate than for breast cancer.3 As with breast cancer, the key overall question is whether the problem lies with fat itself, or with any of the large set of dietary factors that covary with fat intake. The paper by Lee et al. the broadest in this group is distinguished as the first case-control study on prostate cancer to be conducted in China, a country characterized until very recently by extremely low prostate cancer incidence and low fat intake. The results generally confirm the association between high fat diet patterns and prostate cancer risk observed in most, but not all, Western study populations. This vanguard study also contains hints as to the difficulties of conducting conventional epidemiological research in new environments. Sixty-eight percent of the cases with known staging were extraprostatic at diagnosis an unusually high percentage by Western standards. Cases were nearly four-times more likely than controls to report a prev ous diagnosis of benign prostatic hyperplasia or prostatitis which raises the question as to whether cases had increased access to diagnostic services, a feature that could be associated with Westernized diet. Finally, the total calorie and fat intake levels were remarkably high, even assuming an extraordinary level of physical activity. The challenges involved in transferring food frequency methods for measuring past food group and nutrient intake to a completely new culture while avoiding recall bias should not be underestimated.

The American Cancer Society (ACS) publishes Nutrition and Physical Activity Guidelines to serve as a foundation for its communication, policy, and community strategies and ultimately, to affect dietary and physical activity patterns among Americans. These Guidelines, published every 5 years, are developed by a national panel of experts in cancer research, prevention, epidemiology, public health, and policy, and as such, they represent the most current scientific evidence related to dietary and activity patterns and cancer risk. The ACS Guidelines include recommendations for individual choices regarding diet and physical activity patterns, but those choices occur within a community context that either facilitates or interferes with healthy behaviors. Community efforts are essential to create a social environment that promotes healthy food choices and physical activity. Therefore, this committee presents one key recommendation for community action to accompany the four recommendations for individual choices to reduce cancer risk. This recommendation for community action recognizes that a supportive social environment is indispensable if individuals at all levels of society are to have genuine opportunities to choose healthy behaviors. The ACS Guidelines are consistent with guidelines from the American Heart Association and the American Diabetes Association for the prevention of coronary heart disease and diabetes, as well as for general health promotion, as defined by the Department of Health and Human Services' 2005 Dietary Guidelines for Americans.

The incidence of prostate cancer in the United States has recently undergone dramatic and unprecedented changes. Exposure to prostate-specific antigen testing has led to a steep increase in reported incidence. The data indicate that the strongest risk factors for prostate cancer are age and African American race/ethnicity. Family history is also an important risk factor for prostate cancer, although only a small proportion of cases will be due to high-penetrance genes such as those at the putative susceptibility loci (eg, ELAC2) recently identified through linkage analysis. International variation in the risk of prostate cancer is profound, as is the frequency with which migrants from low- to high-risk areas adopt the risk pattern of the host country, possibly within a single generation. This article reviews the current state of knowledge regarding risk factors for prostate cancer, including factors related to diet, anthropometrics, hormone profiles, and concomitant medical conditions. The need for unifying, overarching hypotheses is emphasized, along with identification of some of the barriers to future progress in the field.

Increasing evidence supports the important role of nutrition in cancer prevention, including prevention of prostate cancer. In this review, we summarize data for some of the most consistently observed dietary associations for prostate cancer incidence, briefly consider possible postdiagnostic effects of nutrition on prostate cancer progression/survival, discuss new but limited data on diet-gene interactions, and comment on current areas of controversy for future research focus. Potential protective dietary elements include tomatoes/lycopene, other carotenoids, cruciferous vegetables, vitamin E, selenium, fish/marine omega-3 fatty acids, soy, isoflavones and polyphenols; whereas milk, dairy, calcium, zinc at high doses, saturated fat, grilled meats, and heterocyclic amines may increase risk. It is important to note that randomized clinical trial data exist only for vitamin E, calcium, beta-carotene, and selenium (all of which suggest inverse or no association). Several genes, such as MnSOD, XRCC1, and GST, may modify the association of specific nutrients and foods with prostate cancer risk; and further research is warranted to confirm these initial observed relationships. Until further clinical trial data are available on specific supplements and prostate cancer prevention, it would be prudent to emphasize a diet consisting of a wide variety of plant-based foods and fish; this is similar to what is recommended (and what is more well established) for the primary prevention of heart disease.