A novel panel of blood-based microRNAs capable of discrimination between benign breast disease and breast cancer at early stages.

Breast cancer (BC) as a leading cause of cancer death among women, exhibits a wide range of genetic heterogeneity in affected individuals. Satisfactory management of BC depends on early diagnosis and proper monitoring of patients' response to therapy. In this study, we aimed to assess the relation between the expression patterns of blood-based microRNAs (miRNAs) with demographic characteristics of the patients with BC in an attempt to find novel diagnostic markers for BC with acceptable precision in clinical applications. To this end, we performed comprehensive statistical analysis of the data of the Cancer Genome Atlas (TCGA) database and the blood miRNome dataset (GSE31309). As a result, 21 miRNAs were selected for experimental verification by quantitative RT-PCR on blood samples of 70 BC patients and 60 normal individuals (without any lesions or benign breast diseases). Statistical one-way ANOVA revealed no significant difference in the blood levels of the selected miRNAs in BC patients compared to any lesions or benign breast diseases. However, the multi-marker panel consisting of hsa-miR-106b-5p, -126-3p, -140-3p, -193a-5p, and -10b-5p could detect early-stages of BC with 0.79 sensitivity, 0.86 specificity and 0.82 accuracy. Furthermore, this multi-marker panel showed the potential of detecting benign breast diseases from BC patients with 0.67 sensitivity, 0.80 specificity, and 0.74 accuracy. In conclusion, these data indicate that the present panel might be considered an asset in detecting benign breast disease and BC.

Breast Cancer Risk Factors and Circulating Anti-Müllerian Hormone Concentration in Healthy Premenopausal Women.

CONTEXT: We previously reported that anti-Müllerian hormone (AMH), a marker of ovarian reserve, is positively associated with breast cancer risk, consistent with other studies. OBJECTIVE: This study assessed whether risk factors for breast cancer are correlates of AMH concentration. METHODS: This cross-sectional study included 3831 healthy premenopausal women (aged 21-57, 87% aged 35-49) from 10 cohort studies among the general population. RESULTS: Adjusting for age and cohort, AMH positively associated with age at menarche (P < 0.0001) and parity (P = 0.0008) and inversely associated with hysterectomy/partial oophorectomy (P = 0.0008). Compared with women of normal weight, AMH was lower (relative geometric mean difference 27%, P < 0.0001) among women who were obese. Current oral contraceptive (OC) use and current/former smoking were associated with lower AMH concentration than never use (40% and 12% lower, respectively, P < 0.0001). We observed higher AMH concentrations among women who had had a benign breast biopsy (15% higher, P = 0.03), a surrogate for benign breast disease, an association that has not been reported. In analyses stratified by age (<40 vs ≥40), associations of AMH with body mass index and OCs were similar in younger and older women, while associations with the other factors (menarche, parity, hysterectomy/partial oophorectomy, smoking, and benign breast biopsy) were limited to women ≥40 (P-interaction < 0.05). CONCLUSION: This is the largest study of AMH and breast cancer risk factors among women from the general population (not presenting with infertility), and it suggests that most associations are limited to women over 40, who are approaching menopause and whose AMH concentration is declining.

Evaluation of color Doppler ultrasound combined with plasma miR-21 and miR-27a in the diagnosis of breast cancer.

PURPOSE: To evaluate the clinical value of color Doppler ultrasound combined with plasma miR-21 and miR-27a in the diagnosis of breast cancer (BC). METHODS: 129 patients with BC and 50 patients with benign breast lesions were selected. They were scanned by color Doppler ultrasound on the breast. The relative expression of miR-21 and miR-27a in plasma was detected by real-time fluorescence quantitative PCR. The clinical value of color Doppler ultrasound combined with plasma miR-21 and miR-27a in the diagnosis of BC was analyzed retrospectively. RESULTS: Two-dimensional ultrasound images of BC showed irregular shape, unclear boundary, burr-like edge, uneven internal echo, weakened rear echo, microcalcification, aspect ratio > 1 and axillary lymph node metastasis. The grade of blood flow signal was improved, mainly in grade II and above, which was statistically different from that of benign control group (P < 0.001). The relative expression of plasma miR-21 and miR-27a in breast cancer group was significantly higher than that in benign control group (P < 0.001). The relative expressions of plasma miR-21 and miR-27a in breast cancer group were significantly correlated with histological grade, clinical stage, lymph node metastasis and postoperation. The relative expression levels of miR-21 and miR-27a in plasma of patients with low histological grade (grade III), high clinical stage (stage III, IV) and lymph node metastasis before operation were significantly higher than those in patients with high histological grade (grade I, II), low clinical stage (stage I, II) and no lymph node metastasis after operation (P < 0.01). The sensitivity, accuracy and negative predictive value of ultrasound combined with plasma miR-21 and miR-27a in the diagnosis of BC were significantly higher than those of each single examination. CONCLUSIONS: Color Doppler ultrasound combined with plasma miR-21 and miR-27a can significantly improve the diagnostic efficiency of BC, which is beneficial to early diagnosis and early clinical intervention.

Chinese massage, Tui Na, combined with herbs improves clinical symptoms and regulates sex hormones in patients with mammary gland hyperplasia.

To study the effects of Tui Na therapy on patients with mammary gland hyperplasia.A total of 68 female patients with mammary gland hyperplasia were included in this retrospective study from May 2016 to May 2017 and assigned into control group (N = 34) treated with Rupixiao only (a proprietary Chinese medicine) or Tui Na group (N = 34) treated with Tui Na (Chinese massage) combined with Rupixiao. The pain intensity (visual analogous scale, VAS) and serum levels of luteinizing hormone (LH), estradiol (E2), prolactin (PRL), and progesterone (P) were examined before and after the treatment.The efficacies were 94.1% (32/34) in the Tui Na group and 76.5% (26/34) in the control group (P = .04). After treatment, VAS in Tui Na groups was significantly lower than that in control group (2.1 ±â€Š1.1 vs 3.1 ±â€Š1.1, P < .05). After follow-up for five months, the recurrence rates were 12.5% (4/32) in the Tui Na group and 23.1% (6/26) in the control group (P = .01). The levels of all 4 hormones in the Tui Na group increased significantly after treatment. In control group, only LH and E2 levels were significantly increased after treatment.In patients with mammary gland hyperplasia, Tui Na combined with Rupixiao could improve clinical symptoms, regulate sex hormone levels, and decrease the recurrence rate than Rupixiao alone. Our finding suggests that Tui Na can be potentially used for the treatment of mammary gland hyperplasia.

Correlation of nipple eczema in pregnancy with atopic dermatitis in Northern India: a study of 100 cases.

BACKGROUND: Nipple eczema is a less common presentation of atopic dermatitis. No studies in the literature have correlated nipple eczema in pregnancy as a manifestation of atopic dermatitis. OBJECTIVE: To evaluate whether nipple eczema presenting in pregnancy is a manifestation of atopic dermatitis. METHODS: This was a prospective observational study including 100 women who presented with nipple eczema for the first time during pregnancy. The exclusion criteria were any patient with previous history of nipple eczema, those already on oral or topical treatment for atopic dermatitis or nipple eczema, and other disorders mimicking eczema. Patients were divided into two groups ‒ nipple eczema with atopic dermatitis and without atopic dermatitis. Demographic data, clinical features, total leukocyte count, differential leukocyte count, absolute eosinophil counts, and serum IgE levels were compared between the two groups to detect association between nipple eczema in pregnancy and atopic dermatitis. RESULTS: Out of 100 patients, 39 were diagnosed with atopic dermatitis, whereas 61 were ruled out to have any features suggestive of atopic dermatitis. There were no statistically significant differences in mean age, mean duration of symptoms, and serum IgE levels. In patients with atopic dermatitis, bilateral symptoms were noted more commonly than in patients without the disease, but this was statistically insignificant. STUDY LIMITATIONS: Lack of long term follow-up and no large studies in literature to compare results. CONCLUSION: Nipple eczema in pregnancy follows a similar pattern as in other age groups. The clinical profile of patients is similar in cases with and without atopic dermatitis.

Correlation of nipple eczema in pregnancy with atopic dermatitis in Northern India: a study of 100 cases

Abstract Background Nipple eczema is a less common presentation of atopic dermatitis. No studies in the literature have correlated nipple eczema in pregnancy as a manifestation of atopic dermatitis. Objective To evaluate whether nipple eczema presenting in pregnancy is a manifestation of atopic dermatitis. Methods This was a prospective observational study including 100 women who presented with nipple eczema for the first time during pregnancy. The exclusion criteria were any patient with previous history of nipple eczema, those already on oral or topical treatment for atopic dermatitis or nipple eczema, and other disorders mimicking eczema. Patients were divided into two groups ‒ nipple eczema with atopic dermatitis and without atopic dermatitis. Demographic data, clinical features, total leukocyte count, differential leukocyte count, absolute eosinophil counts, and serum IgE levels were compared between the two groups to detect association between nipple eczema in pregnancy and atopic dermatitis. Results Out of 100 patients, 39 were diagnosed with atopic dermatitis, whereas 61 were ruled out to have any features suggestive of atopic dermatitis. There were no statistically significant differences in mean age, mean duration of symptoms, and serum IgE levels. In patients with atopic dermatitis, bilateral symptoms were noted more commonly than in patients without the disease, but this was statistically insignificant. Study limitations Lack of long term follow-up and no large studies in literature to compare results. Conclusion Nipple eczema in pregnancy follows a similar pattern as in other age groups. The clinical profile of patients is similar in cases with and without atopic dermatitis.

Obesity and adipocytokines in breast cancer and benign breast disease / Obesidad y adipocitocinas en cáncer de mama y enfermedad mamaria benigna

Background: There is a relationship between obesity and the development of breast cancer; adipocytokines are among the mechanisms related. Objective: To describe the impact of obesity in leptin, adiponectin, resistin and tumor necrosis factor-alpha (TNF-alpha) serum levels in women with breast cancer (BC) and benign breast disease (BBD). Methods: A cross-sectional study was carried out with a sample of 54 individuals divided into two groups: BC (n = 27) and BBD (n = 27). Serum levels of leptin, adiponectin, resistin and TNF-alpha were determined. Body mass index (BMI) was calculated. Statistical analysis included mean, standard deviation, median and interquartile range; the differences between groups were determined by Student´s t test, Mann-Whitney U and Kruskall Wallis test to identify differences between the groups. Results: We didn't find any significant differences related to anthropometric characteristics between BC and BBD groups, or to leptin, adiponectin, resistin and TNF-alpha serum levels (p = NS). We found higher leptin serum levels in obese women with BBD compared with non-obese women with BC (med 22.26 versus 4.34 ng/mL; p = 0.028). Adiponectin serum levels in non-obese women from the BC group were higher than serum levels found in non-obese women with BBD (med 7.10 versus 2.22 ng/mL; p = 0.038) Conclusion: We found higher leptin serum levels and lower adiponectin serum levels in BBD women than BC women in this population with high frequency of obesity.

Introducción: existe relación entre la obesidad y el desarrollo de cáncer de mama (CaM); entre los mecanismos implicados están las adipocitocinas. Objetivo: establecer el efecto de la obesidad sobre los valores séricos de leptina, adiponectina, resistina y el factor de necrosis tumoral-alfa (TNF-alfa) en mujeres con CaM y enfermedad mamaria benigna (EMB). Métodos: estudio transversal analítico. Se determinaron los niveles séricos de leptina, adiponectina, resistina y TNF-alfa, así como el índice de masa corporal (IMC) de 27 mujeres con CaM incidente y 27 con EMB. Se utilizaron promedios, desviaciones estándar, medianas (Me) y rangos intercuartílicos, así como las pruebas t de Student, U de Mann-Whitney y Kruskall-Wallis para identificar diferencias entre los grupos. Resultados: no se encontraron diferencias antropométricas, ni en niveles séricos de TNF-alfa, leptina y resistina entre los grupos (p = NS). Los niveles de leptina de las mujeres con EMB y obesidad fueron significativamente superiores que en las mujeres con CaM no obesas (Me 22.26 frente a 4.34 ng/mL; p = 0.028). Los niveles de adiponectina en mujeres con CaM con IMC sin obesidad fueron mayores que los encontrados en pacientes con EMB con IMC y sin obesidad (Me 7.10 frente a 2.22 ng/mL; p = 0.038). Conclusión: en esta población con elevada frecuencia de obesidad, se encontraron niveles mayores de leptina en mujeres con EMB con respecto a las mujeres con CaM; asimismo, se encontró un patrón inverso en adiponectina.

Serum microRNA as potential biomarker to detect breast atypical hyperplasia and early-stage breast cancer.

AIM: The present study aimed to identify microRNA (miRNA) that can be used for not only detecting early-stage breast cancer (BC) but also diagnosing atypical hyperplasia (AH). MATERIALS & METHODS: RT-qPCR detected the expression levels of miRNAs and receiver operating characteristic curves were constructed to evaluate sensitivity and specificity of the assay. RESULTS: miR-24 and miR-103a were expressed in an upward trend in serum of benign proliferative tumor subjects, while they were downregulated significantly in serum of AH (p < 0.005) and early-stage BC subjects (p < 0.005) with high sensitivity and specificity as compared with controls. Bioinformatics analysis also revealed the potential molecular mechanism through which miR-24 and miR-103a regulate tumorigenesis in BC. CONCLUSION: miR-24 and miR-103a were valuable biomarkers for distinguishing AH and early-stage BC from healthy individuals/benign proliferative tumor patients.

The Antisecretory Factor in Plasma and Breast Milk in Breastfeeding Mothers-A Prospective Cohort Study in Sweden.

Inflammation and infection postpartum threaten the mother and her infant. Human milk provides a defense for the infant, but inflammatory complications like mastitis may lead to the cessation of breastfeeding. Antisecretory factor (AF) has a role in the regulation of secretory processes and inflammation. The objective of the study was to describe AF-levels in plasma and breast milk, and in relation to breast complications. Breastfeeding mothers (n = 95) were consecutively recruited at a Well Baby Clinic in Umeå, Sweden. At inclusion four weeks postpartum, samples of venous blood (10 mL) and breast milk (10 mL) were collected. Active AF was analyzed with ELISA using a monoclonal antibody mAb43, and was detected in all samples of plasma and breast milk with a positive correlation (Spearman coefficient = 0.40, p < 0.001; Pearson correlation = 0.34, p < 0.01). High AF-levels in plasma correlated with high AF-levels in breast milk. The results suggest a co-regulation between active AF in plasma and breastmilk, and/or a local regulation of AF in the breast. Further studies are needed to determine the pathways for the activation of AF-levels in breast milk and plasma.

Circulating Cancer-Associated Macrophage-Like Cells Differentiate Malignant Breast Cancer and Benign Breast Conditions.

BACKGROUND: Blood-based testing can be used as a noninvasive method to recover and analyze circulating tumor-derived cells for clinical use. Circulating cancer-associated macrophage-like cells (CAML) are specialized myeloid cells found in peripheral blood and associated with the presence of solid malignancies. We measured CAMLs prospectively in peripheral blood to ascertain their prevalence, specificity, and sensitivity in relation to breast disease status at clinical presentation. METHODS: We report on two related but separate studies: 1) CellSieve microfilters were used to isolate CAMLs from blood samples of patients with known malignant disease (n = 41). Prevalence and specificity was compared against healthy volunteers (n = 16). 2) A follow-up double-blind pilot study was conducted on women (n = 41) undergoing core-needle biopsy to diagnose suspicious breast masses. RESULTS: CAMLs were found in 93% of known malignant patients (n = 38/41), averaging 19.4 cells per sample, but none in the healthy controls. In subjects undergoing core biopsy for initial diagnosis, CAMLs were found in 88% of subjects with invasive carcinoma (n = 15/17) and 26% with benign breast conditions (n = 5/19). CONCLUSION: These preliminary pilot studies suggest that the presence of CAMLs may differentiate patients with malignant disease, benign breast conditions, and healthy individuals. IMPACT: We supply evidence that this previously unidentified circulating stromal cell may have utility as a screening tool to detect breast cancer in various malignancies, irrespective of disease stage. Cancer Epidemiol Biomarkers Prev; 25(7); 1037-42. ©2016 AACR.

Plasma lipidomics profiling identified lipid biomarkers in distinguishing early-stage breast cancer from benign lesions.

BACKGROUND: Breast cancer is very common and highly fatal in women. Current non-invasive detection methods like mammograms are unsatisfactory. Lipidomics, a promising detection method, may serve as a novel prognostic approach for breast cancer in high-risk patients. RESULTS: According the predictive model, the combination of 15 lipid species had high diagnostic value. In the training set, sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of the combination of these 15 lipid species were 83.3%, 92.7%, 89.7%, and 87.9%, respectively. The AUC in the training set was 0.926 (95% CI 0.869-0.982). Similar results were found in the validation set, with the sensitivity, specificity, PPV and NPV at 81.0%, 94.5%, 91.9%, and 86.7%, respectively. The AUC was 0.938 (95% CI 0.889-0.986) in the validation set. METHODS: Using triple quadrupole liquid chromatography electrospray ionization tandem mass spectrometry, this study was to detect global lipid profiling of a total of 194 plasma samples from 84 patients with early-stage breast cancer (stage 0-II) and 110 patients with benign breast disease included in a training set and a validation set. A binary logistic regression was used to build a predictive model for evaluating the lipid species as potential biomarkers in the diagnosis of breast cancer. CONCLUSIONS: The combination of these 15 lipid species as a panel could be used as plasma biomarkers for the diagnosis of breast cancer.

The prevalence of benign breast diseases in patients with nodular goiter and Hashimoto's thyroiditis.

PURPOSE: The evidence regarding association between thyroid disorders and benign and malignant breast diseases is increasing. The studies exploring the relationship between thyroid and breast diseases usually have focused on thyroid autoimmunity and functions in patients with breast pathologies. In this study, we aimed to reveal the frequency of benign breast conditions in patients with nodular thyroid disease (NTD) and Hashimoto's thyroiditis (HT). METHODS: Seventy-one women with NTD, ninety-five women with HT and seventy-two healthy people as a control group were included in the study. Serum thyroid-stimulating hormone, free triiodothyronine, free thyroxine, and thyroid peroxidase autoantibody levels were measured, and thyroid and breast ultrasonography were performed by a single operator in order to determine thyroid and breast pathology. RESULTS: Benign breast diseases were detected in 54.9% of patients with NTD, in 47.4% of those with HT, and 29.2% of control group in the study (p < 0.01 for nodular and Hashimoto groups versus control group). Simple cyst was found to be the most frequent pathology among benign breast diseases; fibrocystic changes, mixed lesions, benign solid mass, ductal ectasia, and complex cyst followed it. No significant relation was found regarding thyroid function and autoimmunity in patients diagnosed with benign breast disease (p > 0.05). CONCLUSIONS: The results of our study support the association between benign breast diseases and thyroid diseases. An important implication of this finding may be demonstration of the necessity of scanning for potential breast pathology in women with nodular or autoimmune thyroid disease in clinical practice.

Correlation of serum toll like receptor 9 and trace elements with lipid peroxidation in the patients of breast diseases.

Toll-like receptors are recognized as redox sensitive receptor proteins and have been implicated in cellular response to oxidative stress. Altered pro-oxidant-antioxidant balance leads to an increased oxidative damage and consequently play an important role in breast diseases. The study was designed to access the oxidative stress status by quantification of byproducts generated during lipid peroxidation and inadequate trace elements during oxidative damage and its effects on the toll like receptor (TLR) activity in patients of breast diseases. Decreased levels of selenium, copper, zinc, magnesium and iron with elevated levels of malondialdehyde (marker of lipid peroxidation) were accompanied by decreased TLR activity in patients of benign breast diseases as well as breast carcinoma. A similar pattern was observed with the advancement of disease and its subsequent progression in breast carcinoma patients. Results of multinomial regression analysis suggest benign breast disease patients are at higher risk of developing breast cancer with high odds ratio of lipid damage.

Increased serum levels of circulating exosomal microRNA-373 in receptor-negative breast cancer patients.

In this study, we compared the blood serum levels of circulating cell-free and exosomal microRNAs, and their involvement in the molecular subtypes of breast cancer patients. Our analyses on cell-free miR-101, miR-372 and miR-373 were performed in preoperative blood serum of 168 patients with invasive breast cancer, 19 patients with benign breast diseases and 28 healthy women. MicroRNAs were additionally quantified in exosomes of 50 cancer patients and 12 healthy women from the same cohort. Relative concentrations were measured by quantitative TaqMan MicroRNA assays and correlated to clinicopathological risk factors. The concentrations of cell-free miR-101 (p=0.013) and miR-373 (p=0.024) were significantly different between patients with breast cancer and benign tumors. A prevalence of miR-101, miR-372 and miR-373 were found in exosomes. The levels of circulating exosomal (but not cell-free) miR-373 were higher in triple negative than luminal carcinomas (p=0.027). Also, estrogen-negative (p=0.021) and progesterone-negative (p=0.01) tumors displayed higher concentrations of exosomal miR-373 than patients with hormone-receptor positive tumors. Overexpression of miR-373 by transfection of MCF-7 cells showed downregulated protein expression of the estrogen receptor, and inhibition of apoptosis induced by camptothecin. Our data indicate that serum levels of exosomal miR-373 are linked to triple negative and more aggressive breast carcinomas.

Diagnostic and prognostic importance of the neutrophil lymphocyte ratio in breast cancer.

BACKGROUND: The aim of this study was to determine diagnostic and prognostic roles of the neutrophil to lymphocyte ratio (NLR) in breast cancer patients. To date, data are limited on associations of primary breast carcinoma (PBC) and benign proliferative breast disease (BPBD) with preoperative NLR values. MATERIALS AND METHODS: Our study covered of 120 female patients with PBC and 50 with BPBD. Diagnostic values of NLR were estimated using sensitivity, specificity and areas under receiver operating characteristic curves (AUC). RESULTS: NLR values were significantly higher in the PBC patients than in those with BPBD, with an AUC of 0.668 in the PBC case. The optimal cut-off for NLR was 2.96 and this was validated in the testing set, giving a sensitivity and a specificity of 79.7% and 76.2%, respectively, in PBC patients. CONCLUSIONS: Preoperative high NLR is a significant diagnostic predictor of distinction of breast cancer from BPBD and elevated NLR is also an important prognostic marker for primary invasive breast cancer.

A comparison of hormonal profiles between breast cancer and benign breast disease: a case-control study.

BACKGROUND: Benign breast disease (BBD), particularly proliferative BBD, is an established breast cancer risk factor. However, there has been no systematic attempt to compare the hormonal profiles of the two conditions. In a case-control investigation in Athens, Greece, we compared levels of estrogens, testosterone and insulin-like growth factor-1 (IGF-1), as well as their principal binding proteins, between breast cancer patients, women with BBD by histological type (proliferative and nonproliferative) and women with no breast pathology. PATIENTS AND METHODS: We studied 466 women with incident breast cancer, 704 women with BBD and 244 healthy women. We used multiple regression to compare log-transformed serum hormone levels of breast cancer patients with those of healthy women and women with BBD by histological type (proliferative and nonproliferative BBD). RESULTS: The hormonal profile of breast cancer in our study was in line with the generally accepted hormonal profile of this disease, as reported from large cohort studies. Compared with healthy women, breast cancer patients tended to have higher levels of steroid hormones. The evidence was strong for estrone (difference 21.5%, P < 0.001), weaker for testosterone (difference 15.8%, P = 0.07) and weaker still for estradiol (difference 12.0%, P = 0.18). Also compared with healthy women, breast cancer patients had barely higher levels of IGF-1 (difference 2.0%, P = 0.51), but had significantly lower levels of IGF binding protein 3 (IGFBP-3) (difference -6.7%, P = 0.001). Compared with women with BBD, breast cancer patients had nonstatistically significantly lower levels of steroid hormones, but they had higher levels of IGF-1 [difference 5.5%, 95% confidence interval (CI) 0.7% to 10.6%] and lower levels of IGFBP-3 (difference -3.7%, 95% CI -6.7% to -0.7%). Differences were more pronounced when breast cancer patients were contrasted to women with proliferative BBD. CONCLUSIONS: Our findings suggest that high levels of IGF-1 may be an important factor toward the evolution of BBD to breast cancer.

Detection of promoter methylation of tumor suppressor genes in serum DNA of breast cancer cases and benign breast disease controls.

Tumors are capable of shedding DNA into the blood stream. This shed DNA may be recovered from serum or plasma. The objective of this study was to evaluate whether pyrosequencing promoter DNA in a panel of 12 breast cancer-related genes (APC, BRCA1, CCND2, CDH1, ESR1, GSTP1, HIN1, P16, RARß, RASSF1, SFRP1 and TWIST) to measure the degree of methylation would lead to a useful serum-based marker of breast cancer. Serum was obtained from women who were about to undergo a breast biopsy or mastectomy at three hospitals from 1977 to 1987 in Grand Rapids, MI USA. We compared the methylation status of 12 genes in serum DNA obtained from three groups of postmenopausal women (mean age at blood collection: 63.0 y; SD 9.9; range 35-91): breast cancer cases with lymph node-positive disease (n = 241); breast cancer cases with lymph node-negative disease (n = 63); and benign breast disease control subjects (n = 234). Overall, median levels of promoter methylation were low, typically below 5%, for all genes in all study groups. For all genes, median levels of methylation were higher (by 3.3 to 47.6%) in lymph node-positive breast cancer cases than in the controls. Comparing mean methylation level between lymph-node positive cases and controls, the most statistically significant findings, after adjustment of the false-positive rate (q-value), were for TWIST (p = 0.04), SFRP1 (p = 0.16), ESR1 (p = 0.17), P16 (p = 0.19) and APC (p = 0.19). For two of these four genes (TWIST, P16), the median methylation level was also highest in lymph-node positive cases, intermediate in lymph node-negative cases and lowest in the controls. The percent of study subjects with mean methylation scores ≥ 5% was higher among lymph node-positive cases than controls for ten genes, and significantly higher for HIN1 and TWIST (22.0 vs. 12.2%, p = 0.04 and 37.9 vs. 24.5%, p = 0.004, respectively). Despite relatively consistent variation in methylation patterns among groups, these modest differences did not provide sufficient ability to distinguish between cases and controls in a clinical setting.

Contrast-enhanced ultrasound examination of the breast: a literature review.

Angiogenesis is the basis for neoplastic growth in human tissues. Nevertheless, neovascularization may be present both in benign and malignant lesions. Although microvascular density assessment is a useful tool for the study of neoplastic vascularization, it cannot be used on a large scale because of the invasiveness of the method. When contrast-enhanced ultrasonography (CEUS) was introduced in clinical practice, the initial results in the field of breast lesions were disappointing because differentiation between benign and malignant masses was not possible. Magnetic resonance imaging (MRI) assessment of the microvascular features was therefore immediately accepted because of the higher diagnostic accuracy. However, in the last decade the sensitivity and specificity of CEUS have greatly improved due to the development of more sophisticated ultrasound (US) equipment, the introduction of second-generation contrast agents and the development of dedicated software able to perform quantitative analysis. This literature review compares the main results reported in the literature regarding the use of CEUS for the characterization of neoplastic lesions of the breast. All the authors agreed that malignant lesions show early wash-in with more intense enhancement and fast wash-out in comparison with benign masses. However, there is still no observer agreement regarding vascularization patterns, and different classifications are proposed. The conclusion of this literature review is therefore that the clinical role of CEUS in the diagnostic process and in follow-up is still to be clearly defined.

Insulin-like growth factor-1, insulin-like growth factor binding protein-3 and lobule type in the Nurses' Health Study II.

INTRODUCTION: Previous research in the Nurses' Health Study (NHS) and the NHSII observed that, among women diagnosed with benign breast disease (BBD), those with predominant type 1/no type 3 lobules (a marker of complete involution) versus other lobule types were at lower risk of subsequent breast cancer. Studies in animal models suggest that insulin-like growth factor-1 (IGF-1) may inhibit involution of lobules in the breast; however, this has not been studied in humans. METHODS: We conducted a cross-sectional study among 472 women in the NHSII who were diagnosed with biopsy-confirmed proliferative BBD between 1991 and 2002 and provided blood samples between 1996 and 1999. A pathologist, blinded to exposure status, classified lobule type in normal adjacent tissue on available biopsy slides according to the number of acini per lobule. For each participant, the pathologist determined the predominant lobule type (that is, type 1, type 2, or type 3) and whether any type 1 or any type 3 lobules were present. Lobule type was then classified as: predominant type 1/no type 3 lobules, which is suggestive of complete involution; or other lobule types. Multivariate logistic models were used to assess the associations between plasma IGF-1, insulin-like growth factor binding protein-3 (IGFBP-3), and the ratio of IGF-1:IGFBP-3 levels with lobule type. RESULTS: In univariate analyses, greater age, higher body mass index, postmenopausal status, nulliparity, and lower IGF-1 levels were associated with predominant type 1/no type 3 lobules (P < 0.05). In multivariate models adjusting for age and assay batch, higher IGF-1 levels were associated with decreased odds of predominant type 1/no type 3 lobules (odds ratio quartile 4 vs. quartile 1 = 0.37, 95% confidence interval = 0.15 to 0.89). Greater ratios of IGF-1:IGFBP-3 levels were also associated with decreased odds of predominant type 1/no type 3 lobules (odds ratio quartile 4 vs. quartile 1 = 0.26, 95% confidence interval = 0.11 to 0.64). These results were slightly attenuated after adjustment for other potential predictors of lobule type. CONCLUSIONS: Higher IGF-1 levels and a greater IGF-1:IGFBP-3 ratio were associated with decreased odds of having predominant type 1 lobules/no type 3 lobules among women with proliferative BBD in the NHSII. This study provides further evidence for the role of insulin-like growth factors in the structure of breast lobules and lobular involution.