Establishment and comparison of two methods to produce a rat model of mammary gland hyperplasia with hyperprolactinemia

Abstract This study aimed to establish and compare models of mammary gland hyperplasia (MGH) with hyperprolactinemia (HPRL) using two different methods. The models provide information on the relationship between mammary gland hyperplasia and associated hormones. Model A was constructed using intramuscular injections of estradiol benzoate injection (EBI), followed by progesterone (P), and then metoclopramide dihydrochloride (MDI). Model B was designed by administering MDI, follow by EBI, and then P intramuscularly. Model B showed higher MGH progression compared with model A. Notably, increase in estradiol (E2) was negatively correlated with prolactin (PRL) secretion. However, PRL levels in model B were significantly higher compared with the levels in model A. Estrogen (ER), prolactin receptor (PRLR), and progesterone receptor (PR) mRNA and protein expression levels in model B rats were positively correlated with changes in the corresponding hormone levels. However, E2, P, and PRL levels in model A showed no direct relationship with levels of the mRNAs of related hormones and protein expression levels. Our results suggest that model B is an appropriate model of MGH with HPRL that can be used to perform further studies about the interactions of the E2, P, and PRL hormones in this disorder.

Three-dimensional microanatomy of human nipple visualized by X-ray dark-field computed tomography.

PURPOSE: The three-dimensional (3D) structure of the human nipple has not been fully clarified. However, its importance has increased in recent years because it has become common practice to preoperatively explore the spread of breast cancer to the nipple with needle biopsy, ductoscopy, and/or ductal lavage for nipple-sparing mastectomy. Here, we demonstrated that X-ray dark-field computed tomography (XDFI-CT) is a powerful tool for reconstructing the 3D distribution pattern of human lactiferous ducts non-destructively, without contrast agent, and with high tissue contrast. METHODS: Nipples amputated from mastectomy specimens of 51 patients with breast cancer were visualized three-dimensionally by XDFI-CT. First, CT images and conventionally stained tissue sections were compared to demonstrate that XDFI-CT provides 3D anatomical information. Next, the number of ducts in the nipple and the number of ducts sharing an ostium near the tip of the nipple were measured from the volume set of XDFI-CT. Finally, the 3D distribution pattern of the ducts was determined. RESULTS: XDFI-CT can provide images almost equivalent to those of low-magnification light microscopy of conventional hematoxylin-eosin-stained histological sections. The mean number of ducts in all cases was 28.0. The total number of ducts sharing an ostium near the tip of the nipple was 525 of 1428. The 3D distribution patterns of the ducts were classified into three types that we defined as convergent (22%), straight (39%), or divergent (39%). CONCLUSIONS: XDFI-CT is useful for exploring the microanatomy of the human nipple and might be used for non-invasive nipple diagnosis in the future.

Assessment of breast self- examination practice and associated factors among female workers in Debre Tabor Town public health facilities, North West Ethiopia, 2018: Cross- sectional study.

BACKGROUND: Although breast Self-Examination is no longer tenable as a standard method to detect early breast cancer, world health organization recommends breast self -examination for raising awareness of women about breast cancer. Secondary prevention through monthly breast self-examination is the best option to tackle the rising incidence of breast cancer. Therefore, the aim of this study was to assess breast self -examination practice and associated factors. METHODS: This cross-sectional study was conducted from April 23 to May 23, 2018. A total of 421 female workers in Debre Tabor Town public health facilities were included. The study participants were selected using simple random sampling technique from the study population. The collected data were checked for completeness. The data were entered and cleaned using EpiData version 3.1 then exported to SPSS version 20 for analysis. Crude odd ratio and probability value were identified for each independent variable and all independent variables with probability value of less than 0.2 were entered into multivariables logistic regression. Statistically significant associated factors were identified based on probability value (p-value) less than 0.05 and adjusted odd ratio with 95% confidence interval. RESULT: The mean age of participants was 25.2 (S.D = 4.12) and 137 (32.5%) of the participants had practiced breast self -examination and 64 (15.2%) of them performed it monthly. Family history of breast cancer (adjusted OR = 6.5, CI = 1.54-21.4), Knowledge about breast -self examination (adjusted OR = 5.74, CI = 2.3-14.4) and self- efficacy in practicing breast self -examination (adjusted OR = 4.7, CI = 1.84-12.11) were significantly associated with breast self -examination practice. CONCLUSIONS: The study showed that the prevalence of breast self-examination was low. Family history of breast cancer, knowledge about breast self -examination and self- efficacy in practicing breast self- examination did have statistically significant association with breast self-examination practice.

Oncoplastic breast surgery and radiotherapy-Adverse aesthetic outcomes, proposed classification of aesthetic components, and causality attribution.

The aim of this paper is to discuss and understand the interplay of surgical and radiotherapy (RT) techniques that potentially affect the aesthetic outcomes following breast conservation treatment. It reviews the reported cosmetic outcomes within the practice-changing RT trials incorporating varied fractionation regimes as well as sequence (before, during, or after surgery) over the past two decades. The paper importantly highlights an overall improvement in breast surgical techniques particularly oncoplastic breast conservation techniques due to dedicated and streamlined training of late. Hence, erstwhile complications or sequel of breast surgery and associated post-RT aesthetic concerns may not be of the same variety or magnitude that were noted in the past. Paper thus concludes by highlighting the paradigm shift in the breast surgical practice that will have a bearing on the accuracy of RT outcomes assessed (within or out with clinical trials) with implications as a potential research and outcomes assessment gap.

Ultrasound Characteristics of the Hair Follicles and Tracts, Sebaceous Glands, Montgomery Glands, Apocrine Glands, and Arrector Pili Muscles.

OBJECTIVES: To explore the capability of very high-frequency ultrasound (US; 50-71 MHz) to detect the normal morphologic characteristics of the hair follicles and tracts, sebaceous glands, Montgomery glands, apocrine glands, and arrector pili muscles. METHODS: A retrospective study, approved by the Institutional Review Board, evaluated the normal US morphologic characteristics of the hair and adnexal structures in a database of very high-frequency US images extracted from the perilesional or contralateral healthy skin of 1117 consecutive patients who underwent US examinations for localized lesions of the skin and 10 healthy individuals from December 2017 to June 2018. These images were matched with their counterparts from the database of normal histologic images according to the corporal region. The Cohen concordance test and regional mean diameters of the hair follicles and adnexal structures were analyzed. RESULTS: The normal hair follicles and tracts, sebaceous glands, Montgomery glands, apocrine glands, and arrector pili muscles were observed on US images and matched their histological counterparts in all the corporal regions. There was significant US concordance (κ = 0.82; P = .0001) among observers. Regional mean diameters (millimeters) of the hair follicles, sebaceous glands, and apocrine glands are provided. CONCLUSIONS: The hair follicles and tracts, sebaceous glands, Montgomery glands, apocrine glands, and arrector pili muscles are detectable with very high-frequency US, including some regional and anatomic variants. Knowledge of their normal US appearances is a requisite for detecting subclinical changes, understanding the physiopathologic characteristics, and supporting the early diagnosis and management of common dermatologic diseases.

PVCP-based anthropomorphic breast phantoms containing structures similar to lactiferous ducts for ultrasound imaging: A comparison with human breasts.

The purpose of this work was to obtain an anthropomorphic phantom with acoustic properties similar to those of breast tissue, possessing lactiferous duct-like structures, which would be a first for this type of phantom. Breast lesions usually grow in glandular tissues or lactiferous ducts. Shape variations in these structures are detectable by using ultrasound imaging. To increase early diagnosis, it is important to develop computer-aided diagnosis (CAD) systems and improve medical training. Using tissue-like materials that mimic known internal structures can help achieve both of these goals. However, most breast ultrasound phantoms described in the literature emulate only fat tissues and lesion-like masses. In addition, commercially available phantoms claim to be realistic, but do not contain lactiferous duct structures. In this work, we collected reference images from both breasts of ten healthy female volunteers aged between 20 and 30 years using a 10 MHz linear transducer of a B-mode medical ultrasound system. Histograms of the grey scale distribution of each tissue component of interest, the grey level means, and standard deviations of the regions of interest were obtained. Phantoms were produced using polyvinyl chloride plastisol (PVCP) suspensions. The lactiferous duct-like structures were prepared using pure PVCP. Solid scatterers, such as alumina (mesh #100) and graphite powders (mesh #140) were added to the phantom matrix to mimic glandular and fat tissue, respectively. The phantom duct-like structure diameters observed on B-mode images (1.92 mm ±â€¯0.44) were similar to real measures obtained with a micrometer (2.08 mm ±â€¯0.23). The phantom ducts are easy to produce and are largely stable for at least one year. This phantom allows the researchers to elaborate the structure at their will and may be used in training and as a reference for development of CAD systems.

Environmental selection during the last ice age on the mother-to-infant transmission of vitamin D and fatty acids through breast milk.

Because of the ubiquitous adaptability of our material culture, some human populations have occupied extreme environments that intensified selection on existing genomic variation. By 32,000 years ago, people were living in Arctic Beringia, and during the Last Glacial Maximum (LGM; 28,000-18,000 y ago), they likely persisted in the Beringian refugium. Such high latitudes provide only very low levels of UV radiation, and can thereby lead to dangerously low levels of biosynthesized vitamin D. The physiological effects of vitamin D deficiency range from reduced dietary absorption of calcium to a compromised immune system and modified adipose tissue function. The ectodysplasin A receptor (EDAR) gene has a range of pleiotropic effects, including sweat gland density, incisor shoveling, and mammary gland ductal branching. The frequency of the human-specific EDAR V370A allele appears to be uniquely elevated in North and East Asian and New World populations due to a bout of positive selection likely to have occurred circa 20,000 y ago. The dental pleiotropic effects of this allele suggest an even higher occurrence among indigenous people in the Western Hemisphere before European colonization. We hypothesize that selection on EDAR V370A occurred in the Beringian refugium because it increases mammary ductal branching, and thereby may amplify the transfer of critical nutrients in vitamin D-deficient conditions to infants via mothers' milk. This hypothesized selective context for EDAR V370A was likely intertwined with selection on the fatty acid desaturase (FADS) gene cluster because it is known to modulate lipid profiles transmitted to milk from a vitamin D-rich diet high in omega-3 fatty acids.

Time to redefine the intramammary lymph node as a separate entity?

The lymphatic drainage for the majority of primary breast tumors is to the axillary lymph nodes (ALNs). Some, however, drain to the so-called extra-axillary basins, namely the internal mammary, supra- and infraclavicular regions. Another potential drainage route includes the intramammary lymph nodes (IMLNs). Current guidance suggests IMLNs should be considered as part of the axillary group, potentially affecting axillary management. However, due to evolution in imaging and advancement in technology, IMLNs may now be distinguished more accurately pre-operatively. There are currently no published guidelines for the management of IMLNs in the United Kingdom. The authors suggest that it is time to reclassify IMLNs as a separate focus of cancer and treat it as a separate entity. Clin. Anat. 31:684-687, 2018. © 2018 Wiley Periodicals, Inc.

Breast education for schoolgirls; why, what, when, and how?

Schools are fundamental settings for health education and adolescent females are an important group for promoting positive breast habits. We surveyed 2089 schoolgirls (11-18 years) to provide evidence for, and guidance on, breast education for schoolgirls. 26% reported negative feelings about their breasts and 87% reported ≥ one breast concern. 72% wanted to know more about breast cancer (69% rating this extremely important). >50% wanted to know more about breast sag and breast pain. Preferred delivery format was age eleven (50%), girls only taught sessions (41%) with female teachers (43%). A need for breast education and delivery preferences was identified.

Image analysis of immune cell patterns in the human mammary gland during the menstrual cycle refines lymphocytic lobulitis.

PURPOSE: To improve microscopic evaluation of immune cells relevant in breast cancer oncoimmunology, we aim at distinguishing normal infiltration patterns from lymphocytic lobulitis by advanced image analysis. We consider potential immune cell variations due to the menstrual cycle and oral contraceptives in non-neoplastic mammary gland tissue. METHODS: Lymphocyte and macrophage distributions were analyzed in the anatomical context of the resting mammary gland in immunohistochemically stained digital whole slide images obtained from 53 reduction mammoplasty specimens. Our image analysis workflow included automated regions of interest detection, immune cell recognition, and co-registration of regions of interest. RESULTS: In normal lobular epithelium, seven CD8[Formula: see text] lymphocytes per 100 epithelial cells were present on average and about 70% of this T-lymphocyte population was lined up along the basal cell layer in close proximity to the epithelium. The density of CD8[Formula: see text] T-cell was 1.6 fold higher in the luteal than in the follicular phase in spontaneous menstrual cycles and 1.4 fold increased under the influence of oral contraceptives, and not co-localized with epithelial proliferation. CD4[Formula: see text] T-cells were infrequent. Abundant CD163[Formula: see text] macrophages were widely spread, including the interstitial compartment, with minor variation during the menstrual cycle. CONCLUSIONS: Spatial patterns of different immune cell subtypes determine the range of normal, as opposed to inflammatory conditions of the breast tissue microenvironment. Advanced image analysis enables quantification of hormonal effects, refines lymphocytic lobulitis, and shows potential for comprehensive biopsy evaluation in oncoimmunology.

Melanin of the nipple areola complex.

Biological pigments or biochromes are ubiquitous in animals, plants, and simpler organisms such as fungi and bacteria. They serve a wide spectrum of functions from photosynthesis, camouflage, mimicry, photo protection from the environment to attracting mates. The human female nipple areola complex (NAC) is a highly-pigmented area. Currently, the prevailing theory as to the evolution of the pigmented human NAC is based on infant recognition of breast feeding latching zone; however, due to the protruding shape of the nipple and surrounding breast, the authors of this letter believe that the evolutionary advantage of the pigmented NAC has a direct physiological function, namely the initiation of involution at the end of the infant lactation period.

The need for supplemental breast cancer screening modalities: a perspective of population-based breast cancer screening programs in Japan.

PURPOSE: This article discusses possible supplemental breast cancer screening modalities for younger women with dense breasts from a perspective of population-based breast cancer screening program in Japan. CONCLUSION: Supplemental breast cancer screening modalities have been proposed to increase the sensitivity and detection rates of early stage breast cancer in women with dense breasts; however, there are no global guidelines that recommend the use of supplemental breast cancer screening modalities in such women. Also, no criterion standard exists for breast density assessment. Based on the current situation of breast imaging in Japan, the possible supplemental breast cancer screening modalities are ultrasonography, digital breast tomosynthesis, and breast magnetic resonance imaging. An appropriate population-based breast cancer screening program based on the balance between cost and benefit should be a high priority. Further research based on evidence-based medicine is encouraged. It is very important that the ethnicity, workforce, workflow, and resources for breast cancer screening in each country should be considered when considering supplemental breast cancer screening modalities for women with dense breasts.

Morphological and neuroanatomical study of the mammary gland in the immature and mature European beaver (Castor fiber).

This study investigated general morphology and immunohistochemical properties of nerve fibres supplying the mammary gland (MG) in the European beaver. The microscopic analysis of the beaver mammary gland revealed the presence of morphological structures which are characteristic for mammals. There were no distinct differences in the morphological features of the mammary gland between the juvenile and non-pregnant mature beaver. The nerve fibres were visualized using antibodies against protein gene product 9.5 (PGP) and biologically active substances including ß-hydroxylase tyrosine (DßH), neuropeptide Y (NPY), calcitonine gene-related peptide (CGRP) and substance P (SP). The study has revealed that the MG in the juvenile and mature beaver is richly supplied with PGP-immunoreactive (PGP-IR) nerve fibres. The most abundant innervation was observed in the nipple and less numerous nerve terminals supplied the glandular tissue. Double-labelling immunohistochemistry disclosed that the majority of PGP-IR nerve fibres associated with blood vessels and smooth muscle cells in both the nipple and glandular tissue were also DßH-IR. However, these nerve terminals were less numerous in the glandular tissue than in the nipple. Most of the DßH-IR axons associated with arteries and smooth muscle cells in the entire gland also stained for NPY. Small number of DßH/NPY-IR fibres supplied veins. CGRP-IR fibres were more abundant than those expressing SP. No distinct differences in the distribution and immunohistochemical characteristic of nerve fibres were observed between the juvenile and adult animals. The distribution and immunohistochemical properties of nerve fibres supplying the gland in the beaver remind those previously described in other mammalian species.

Ages at menarche- and menopause-related genetic variants in relation to terminal duct lobular unit involution in normal breast tissue.

Reduced levels of terminal duct lobular unit (TDLU) involution, as reflected by higher numbers of TDLUs and acini per TDLU, have been associated with higher breast cancer risk. Younger age at menarche and older age at menopause have been previously related to lower levels of TDLU involution. To determine a possible genetic link, we examined whether single-nucleotide polymorphisms (SNPs) previously established in genome-wide association studies (GWAS) for ages at menarche and menopause are associated with TDLU involution. We conducted a pooled analysis of 862 women from two studies. H&E tissue sections were assessed for numbers of TDLUs and acini/TDLU. Poisson regression models were used to estimate associations of 36 menarche- and 21 menopause-SNPs with TDLU counts, acini counts/TDLU, and the product of these two measures, adjusting for age and study site. Fourteen percent of evaluated SNPs (eight SNPs) were associated with TDLU counts at p < 0.05, suggesting an enrichment of associations with TDLU counts. However, only menopause-SNPs had >50 % that were either significantly or nonsignificantly associated with TDLU measures in the directions consistent with their relationships shown in GWAS. Among ten SNPs that were statistically significantly associated with at least one TDLU involution measure (p < 0.05), seven SNPs (rs466639: RXRG; rs2243803: SLC14A2; rs2292573: GAB2; rs6438424: 3q13.32; rs7606918: METAP1D; rs11668344: TMEM150B; rs1635501: EXO1) were associated in the consistent directions. Our data suggest that the loci associated with ages at menarche and menopause may influence TDLU involution, suggesting some shared genetic mechanisms. However, larger studies are needed to confirm the results.

[Estimation of the Average Glandular Dose Using the Mammary Gland Image Analysis in Mammography].

Currently, the glandular dose is evaluated quantitatively on the basis of the measured data using phantom, and not in a dose based on the mammary gland structure of an individual patient. However, mammary gland structures of the patients are different from each other and mammary gland dose of an individual patient cannot be obtained by the existing methods. In this study, we present an automated estimation method of mammary gland dose by means of mammary structure which is measured automatically using mammogram. In this method, mammary gland structure is extracted by Gabor filter; mammary region is segmented by the automated thresholding. For the evaluation, mammograms of 100 patients diagnosed with category 1 were collected. Using these mammograms we compared the mammary gland ratio measured by proposed method and visual evaluation. As a result, 78% of the total cases were matched. Furthermore, the mammary gland ratio and average glandular dose among the patients with same breast thickness was matched well. These results show that the proposed method may be useful for the estimation of average glandular dose for the individual patients.

Oncoplastic surgery combining abdominal advancement flaps with volume displacement techniques to breast-conserving surgery for small- to medium-sized breasts.

BACKGROUND: An abdominal advancement flap (AAF) is a flap that pulls the elevated abdominal skin up, creating the shape of the inferior portion of the breast by making a neo-inframammary fold. We used an AAF combined with volume displacement techniques to fill the defect left after breast-conserving surgery (BCS). METHODS: Forty-one small- to medium-sized breast patients whose resection area included the lower portion of the breast underwent this procedure from October 2010 to December 2014. We evaluated efficacy of this procedure. RESULTS: The excision volume ranged from 10 to 35 %. Complications after surgery were observed in two patients (partial necrosis of the nipple-areola complex and partial necrosis of the breast skin in one patient each). There was no fat necrosis of the flap in any of the patients. The cosmetic results were found to be excellent in 7 cases, good in 23, fair in 9 and poor in 2. In 11 cases with an unacceptable outcome, 9 cases were in the inner portion. In patients with the tumor in the inner portion, the proportion of unacceptable cases was 50 %. In the cases other than the inner portion, the proportion of unacceptable cases was 8.7 % (p < 0.01). In the cases with larger breasts, unacceptable cases were more frequently observed (p < 0.05). CONCLUSIONS: We believe that an AAF combined with volume displacement techniques may be useful following BCS in the lower portion of a small- to medium-sized breast, except in cases where the tumors is located in the inner potion.

Suspensory Ligament of the Mammary Gland: A Case Report.

BACKGROUND: The ligamentous system of the breast comprises a deep component, connecting the breast to the chest wall, and a superficial component, attached to the dermis and defining the breast contour. METHODS: A female patient with a family history of breast and ovarian cancer and presenting a bilateral fibrocystic mastopathy was submitted to surgical treatment with bilateral subcutaneous mastectomy and immediate reconstruction with implants. RESULTS: Intra-operatively, a strong ligament between the clavicle and the upper pole of the breast was identified. In addition, there was bilateral absence of the suspensory ligament of the axilla. CONCLUSIONS: Variations on the anatomy of the ligamentous system of the breast can represent alterations during the embryonic development of the breast and may have an important impact in the development of breast ptosis. LEVEL OF EVIDENCE V: This journal requires that the authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266.

The Cleared Mammary Fat Pad Transplantation Assay for Mammary Epithelial Organogenesis.

Cleared mammary fat pad (MFP) transplantation has been a standard technique for studies of mammary development and cancer for several decades. The mammary gland is comprised of several fundamental components: The epithelial compartment contains basal/myoepithelial cells and luminal cells, and the stromal compartment (called the MFP) contains adipocytes, smooth muscle cells, fibroblasts, and immune cells. In 3- to 4-wk-old female mice, the mammary epithelium is concentrated very close to the nipple and has not yet grown beyond the mammary lymph node to penetrate the bulk of the MFP. This developmental feature provides an anatomical fixed point, and enables one to cut away the portion of the MFP from the nipple to the lymph node, leaving behind the majority of the MFP free of epithelium. The "cleared" MFP can serve as a supportive native microenvironment fully sufficient for the organogenesis of injected donor epithelium. Normal mammary epithelial donor cells will produce histologically and functionally normal mammary ductal epithelium several weeks posttransplant, with the exception that the ducts will not be connected to the nipple. The assay described here provides a powerful platform for assessing the developmental and tumorigenic potential of engineered cells of interest.

Why Does the Weeping Willow Weep? Reconceptualizing Oncogenesis in Breast Cancer.

Like other human anatomical systems, the mammary ducts and glands at the time of lactation adhere to a perfect fractal geometry - which is destroyed by breast cancer. Could a loss of fractal geometry of the blood supply to human tissue be an early step in oncogenesis?

Breastmilk Expression and Breast Pump Technology.

More than 80% of mothers express milk at some point during the first 4 months postpartum. Many women rely on breast pumps to establish and maintain lactation particularly when the infant is not able to breastfeed. Proper breast pump fit and use are important to optimize breastmilk supply and prevent injury. As breast pumps continues to evolve and the number of women using them remains a substantial majority, health care professionals need to maintain a thorough understanding of this tool and relevant techniques to help mothers succeed.