Computerized Chronothermodynamic Breast Examinations Under Ambulatory Conditions

A new approach to the assessment of breast health was developed based upon continuous ambulatory recording of breast temperature for 24 hrs. A new data acquisition system consisting of a portable, multi-channel, microprocessor-based recorder worn on a belt was developed to record temperatures. Dramatic thermal fluctuations were noted during the recordings in response to multiple stimuli (physiological, behavioral, environmental). Specific algorithms and software programs were developed to process the thermobiologic and chronobiologic information. Data processing was fully automatic and included an examination report indicating a score and a chronothermodynamic class for each of the five areas examined on each breast. The preliminary clinical studies have concerned more than 450 patients with healthy or diseased breasts.     

Development and characterization of breast carcinoma cell lines as in vitro and in vivo models for breast cancer diagnosis and therapy

Summary To establish a model system for preclinical radioimmunotherapy studies, attempts were made to graft 16 different human breast carcinoma cell lines into BALB/c nu/nu (nude) mice. Nine produced serially transplantable tumors growing at a variable rate, whereas seven failed to do so. Conversely, three new cell lines were established in monolayer culture from transplantable human breast tumors in nude mice. Twelve selected tumors and their corresponding cell lines were characterized for DNA ploidy, % S-phase, and breast epithelial mucin expression by immunohistochemistry and flow cytometry. A wide diversity of these cellular characteristics were found in that each tumor was unique and distinct from the others. DNA ploidy differed among the tumors but was not affected by switching between in vitro to in vivo growth. Some tumors expressed similar levels of the breast mucin both in vitro and in vivo, whereas most expressed lower levels as transplantable tumors. There was a good correlation between immunohistochemical and flow cytometric determination of surface and cytoplasmic mucin expression, and with both techniques estrogen and progesterone receptor positive tumors had significantly higher levels of mucin expression than receptor negative tumors. These 12 transplantable breast tumors, with their corresponding cell lines, provide an excellent model system for testing radioimmunotherapy and other therapeutic reagents because they exhibit diverse phenotypic characteristics that represented a mini-population of breast cancer patients’ tumors, allowing assessment of the effect of therapy when confronted with different breast tumors’ genotype and phenotype.     

Development and characterization of new immortalized human breast cell lines

New human breast cell lines were developed from metastatic breast cancer tissues and normal breast tissues. Primary cultures were initiated from cellular outgrowths of explanted tissues or from mechanically isolated cells in two serum-free media. Cell cultures derived from both cancer and normal tissues were immortalized with pRSV-T plasmid to generate permanent breast cell lines that exhibited an epithelial morphology. Cell lines generated in this study were characterized with respect to morphology, growth rate, karyotype, presence of specific genes, and the expression of epithelial and breast markers. The cell lines expressed the epithelial cell markers, cytokeratins 8 and 18, and retained the capacity to produce human milk fat globulin. They also express the BRCA-1, erbB2, and EGF receptor genes and possess the H-ras, K-ras, and p53 genes. Preliminary data showed that one of the new cancer cell lines was highly sensitive to the cytotoxic action of taxol. It is envisioned that the new breast cell lines will be useful as targets for identification of therapeutic agents against breast cancer and as models for carcinogenesis studies. This revised version was published online in June 2006 with corrections to the Cover Date.     

Comparison of dose distributions and organs at risk (OAR) doses in conventional tangential technique (CTT) and IMRT plans with different numbers of beam in left-sided breast cancer

Aim

Our aim was to improve dose distribution to the left breast and to determine the dose received by the ipsilateral lung, heart, contralateral lung and contralateral breast during primary left-sided breast irradiation by using intensity modulated radiotherapy (IMRT) techniques compared to conventional tangential techniques (CTT). At the same time, different beams of IMRT plans were compared to each other in respect to CI, HI and organs at risk (OAR) dose.

Background

Conventional early breast cancer treatment consists of lumpectomy followed by whole breast radiation therapy. CTT is a traditional method used for whole breast radiotherapy and includes standard wedged tangents (two opposed wedged tangential photon beams). The IMRT technique has been widely used for many treatment sites, allowing both improved sparing of normal tissues and more conformal dose distributions. IMRT is a new technique for whole breast radiotherapy. IMRT is used to improve conformity and homogeneity and used to reduce OAR doses.

Materials and methods

Thirty patients with left-sided breast carcinoma were treated between 2005 and 2008 using 6, 18 or mixed 6/18 MV photons for primary breast irradiation following breast conserving surgery (BCS). The clinical target volume [CTV] was contoured as a target volume and the contralateral breast, ipsilateral lung, contralateral lung and heart tissues as organs at risk (OAR). IMRT with seven beams (IMRT7), nine beams (IMRT9) and 11 beams (IMRT11) plans were developed and compared with CTT and among each other. The conformity index (CI), homogeneity index (HI), and doses to OAR were compared to each other.

Results

All of IMRT plans significantly improved CI (CTT: 0.76; IMRT7: 0.84; IMRT9: 0.84; IMRT11: 0.85), HI (CTT: 1.16; IMRT7: 1.12; IMRT9: 1.11; IMRT11: 1.11), volume of the ipsilateral lung receiving more than 20 Gy (>V_20 Gy) (CTT: 14.6; IMRT7: 9.08; IMRT9: 8.10; IMRT11: 8.60), and volume of the heart receiving more than 30 Gy (>V_30 Gy) (CTT: 6.7; IMRT7: 4.04; IMRT9: 2.80; IMRT11: 2.98) compared to CTT. All IMRT plans were found to significantly decrease >V_20 Gy and >V_30 Gy volumes compared to conformal plans. But IMRT plans increased the volume of OAR receiving low dose radiotherapy: volume of contralateral lung receiving 5 and 10 Gy (CTT: 0.0–0.0; IMRT7: 19.0–0.7; IMRT9: 17.2–0.66; IMRT11: 18.7–0.58, respectively) and volume of contralateral breast receiving 10 Gy (CTT: 0.03; IMRT7: 0.38; IMRT9: 0.60; IMRT11: 0.68). The differences among IMRT plans with increased number of beams were not statistically significant.

Conclusion

IMRT significantly improved conformity and homogeneity index for plans. Heart and lung volumes receiving high doses were decreased, but OAR receiving low doses was increased.     

Validation of an optimized method for the determination of iodine in human breast milk by inductively coupled plasma mass spectrometry (ICPMS) after tetramethylammonium hydroxide extraction

In this study a novel method to determine iodine concentrations in human breast milk was developed and validated. The iodine was analyzed by inductively coupled plasma mass spectrometry (ICPMS) following tetramethylammonium hydroxide (TMAH) extraction at 90 °C in disposable polypropylene tubes. While similar approaches have been used previously, this method adopted a shorter extraction time (1 h vs. 3 h) and used antimony (Sb) as the internal standard, which exhibited greater stability in breast milk and milk powder matrices compared to tellurium (Te). Method validation included: defining iodine linearity up to 200 μg L⁻¹; confirming recovery of iodine from NIST 1549 milk powder. A recovery of 94–98% was also achieved for the NIST 1549 milk powder and human breast milk samples spiked with sodium iodide and thyroxine (T4) solutions. The method quantitation limit (MQL) for human breast milk was 1.6 μg L⁻¹. The intra-assay and inter-assay coefficient of variation for the breast milk samples and NIST powder were <1% and <3.5%, respectively. NIST 1549 milk powder, human breast milk samples and calibration standards spiked with the internal standard were all stable for at least 2.5 months after extraction. The results of the validation process confirmed that this newly developed method provides greater accuracy and precision in the assessment of iodine concentrations in human breast milk than previous methods and therefore offers a more reliable approach for assessing iodine concentrations in human breast milk.     

Mapping quantitative trait loci affecting fatness and breast muscle weight in meat-type chicken lines divergently selected on abdominal fatness

Quantitative trait loci (QTL) for abdominal fatness and breast muscle weight were investigated in a three-generation design performed by inter-crossing two experimental meat-type chicken lines that were divergently selected on abdominal fatness. A total of 585 F₂ male offspring from 5 F₁ sires and 38 F₁ dams were recorded at 8 weeks of age for live body, abdominal fat and breast muscle weights. One hundred-twenty nine microsatellite markers, evenly located throughout the genome and heterozygous for most of the F₁ sires, were used for genotyping the F₂ birds. In each sire family, those offspring exhibiting the most extreme values for each trait were genotyped. Multipoint QTL analyses using maximum likelihood methods were performed for abdominal fat and breast muscle weights, which were corrected for the effects of 8-week body weight, dam and hatching group. Isolated markers were assessed by analyses of variance. Two significant QTL were identified on chromosomes 1 and 5 with effects of about one within-family residual standard deviation. One breast muscle QTL was identified on GGA1 with an effect of 2.0 within-family residual standard deviation.     

Quantitation of human mammary epithelial antigens in cells cultured from normal and cancerous breast tissues

Summary A sensitive radioimmunoassay technique was developed to quantitatite the level of human breast celltype specific antigens on cells from normal breast and from various established cell lines of breast and nonbreast origins. Polyacrylamide gel electrophoresis revealed four major proteinaceous components (150,000; 75,000; 60,000; and 48,000) in human milk fat globule membranes that were used to immunize rabbits in order to elicit antimammary epithelial cell antibody. Antisera obtained were rendered specific by abosorptions and were able to recognize three specific mammary epithelial components of the breast epithelial cell. Human mammary epithelial (HME) antigen expression was highest (1290 ng/10⁶ cells) in normal breast epithelial cells from primary cultures of normal breasts. Lower levels (range: 955 to 330 ng/10⁶ cells) were found in breast epithelial cells from cell lines established from cancerous breast tissue. Cells of nonbreast origins as well as fibroblasts from breast gave much lower values (less than 30 ng/10⁶ cells). On treatment, with trypsin, of two breast epithelial cell lines (MDA-MB-157 and MCF-7) 80 to 85% of their HME antigen expression was lost, suggesting that a majority of these breast antigens reside on the cell surface. This work was Supported by Grant PTD-99 from the American Cancer Society, Grant CA19455 and CA20286 from the National Cancer Institute, and Biomedical Research Support Grant RR05467 from the National Institutes of Health. Most cells used in the present study were produced with support from National Cancer Institute Contract Y01-CP8-0500, Biological Carcinogenesis Branch, Division of Cancer Cause and Prevention, under the auspices of the Office of Naval Research and the Regents of the University of California.     

保乳手术治疗92例早期乳腺癌的临床疗效观察

目的：观察保留乳房手术治疗早期乳腺癌的临床治疗效果及其安全性。方法：回顾性分析我院2008年5月-2010年5月间收治的92例行保乳手术的早期乳腺癌患者的,临床资料,观察实施保乳手术并综合后的治疗疗效和患者生活质量变化。结果：全部患者均顺利接受手术,近期乳房外形保持较好,美容效果好,手术并发症少。术后随访20-44个月,1例局部复发,1例肝转移,无死亡病例。结论：保乳手术治疗早期乳腺癌的美容效果好,并发症少,疗效显著;术前严格掌握手术适应症,术后规范的综合治疗,是取得良好效果的基础。     

Bystander killing of breast cancer MCF-7 cells by MDA-MB-231 cells exposed to 5-fluorouracil is mediated via Fas

The major drawback with cancer therapy is the development of resistant cells within tumors due to their heterogeneous nature and due to inadequate drug delivery during chemotherapy. Therefore, the propagation of injury ("bystander effect" (BE)) from directly damaged cells to other cells may have great implications in cancer chemotherapy. The general advantage of the bystander cell killing phenomenon is the large therapeutic index that can be achieved. Experiments suggest that this phenomenon is detected in radiation therapy as well as in gene therapy in conjunction with chemotherapy. In the present study, we developed an original in vitro model dedicated to the exploration of bystander cytotoxicity induced during breast carcinoma chemotherapy. In brief, we investigated this perpetuation of injury on untreated bystander MCF-7 breast cancer cells which were coplated with 5-fluorouracil (5-FU)-treated MDA-MB-231 breast cancer cells. To achieve this goal, a specific in vitro coculture model which involved mixing of aggressive MDA-MB-231 breast cancer cells with enhanced green fluorescent protein (EGFP) expressing stable clone of non-metastatic MCF-7 breast cancer cells (MCF-EGFP), was used. A bystander killing effect was observed in MCF-EGFP cells cocultured with MDA-MB-231 cells pretreated with 5-FU. The striking decrease in MCF-EGFP cells, as detected by assaying for total GFP intensity, is mediated by activation of Fas/FasL system. The implication of Fas in MCF-EGFP cell death was confirmed by using antagonistic anti-FasL antibody that reverses bystander cell death by blocking FasL on MDA-MB-231 cells. In addition, inhibition of CD95/Fas receptor on the cell surface of MCF-EGFP cells by treatment with Pifithrin-alpha, a p53 specific transactivation inhibitor, partially abrogated the sensitivity of bystander MCF-EGFP cells. Our data, therefore, demonstrates that the Fas/FasL system could be considered as a new determinant for chemotherapy-induced bystander cell death in breast cancers.     

保乳手术治疗92 例早期乳腺癌的临床疗效观察

目的：观察保留乳房手术治疗早期乳腺癌的临床治疗效果及其安全性。方法：回顾性分析我院2008 年5 月—2010 年5 月间 收治的92 例行保乳手术的早期乳腺癌患者的临床资料,观察实施保乳手术并综合后的治疗疗效和患者生活质量变化。结果：全 部患者均顺利接受手术,近期乳房外形保持较好,美容效果好,手术并发症少。术后随访20～44 个月,1 例局部复发,1 例肝转移, 无死亡病例。结论：保乳手术治疗早期乳腺癌的美容效果好,并发症少,疗效显著;术前严格掌握手术适应症,术后规范的综合治 疗,是取得良好效果的基础。     

Characterization of the imaging settings in screening mammography using a tracking and reporting system: A multi-center and multi-vendor analysis

A tracking and reporting system was developed to monitor radiation dose in X-ray breast imaging. We used our tracking system to characterize and compare the mammographic practices of five breast imaging centers located in the United States and Brazil. Clinical data were acquired using eight mammography systems comprising three modalities: computed radiography (CR), full-field digital mammography (FFDM), and digital breast tomosynthesis (DBT). Our database consists of metadata extracted from 334,234 images. We analyzed distributions and correlations of compressed breast thickness (CBT), compression force, target-filter combinations, X-ray tube voltage, and average glandular dose (AGD). AGD reference curves were calculated based on AGD distributions as a function of CBT. These curves represent an AGD reference for a particular population and system. Differences in AGD and imaging settings were attributed to a combination of factors, such as improvements in technology, imaging protocol, and patient demographics. The tracking system allows the comparison of various imaging settings used in screening mammography, as well as the tracking of patient- and population-specific breast data collected from different populations.     

Extramammary metastatic neoplasms in the breast: a cytomorphological study of 11 cases

The occurrence of metastatic tumours in the breast is uncommon and it is crucial for cytologists to be aware and distinguish them cytologically from primary breast tumours in fine needle aspirates. In the present retrospective study of 11 cases, over a 20-year period, we discuss the cytological features of extramammary metastatic tumours in the breast. A brief attempt has been made to discuss the past literature. The 11 metastatic tumours included four haematolymphoid neoplasms, two melanomas, two metastatic sarcomas and three metastatic carcinomas. A prior clinical diagnosis of the primary tumour was obtained in seven cases. Immunohistochemistry or histology following a cytological diagnosis confirmed all the cases. The main objective of this study was to highlight the use of cytology and at the same time caution the cytologist to be aware of the clinical/imaging findings and if necessary to utilize immunohistochemical facilities to consider/rule out the possibility of metastatic tumour in the breast.     

利用δ声波场和近红外光实现乳腺肿瘤的精确定位

本文提出一种利用δ声波场和近红外光漫射理论实现球形乳腺肿瘤精确定位的新颖思路。通过构建一个δ波形的声波场,作用到乳腺组织中从而改变组织内某一点的光学特性参数,这种改变对组织表面光分布的影响可以视为微扰,通过控制其作用点在深度上扫描,测得一系列乳腺组织表面一级微扰光分布,从中提取肿瘤与正常组织的差异特性,实现乳腺肿瘤的精确定位和大小测定。该方法具有广泛的临床医学应用前景,为乳腺癌的早期检测提供一种全新思路。     

基于电化学发光检测技术的乳腺癌生物标记物EZH2表达定量分析

Zeste同源染色体2增强子基因(EZH2)在人类乳腺癌中过度表达,它可以被视为一个检测肿瘤的发展和转移的生物标记物。传统技术检测或定量特异性基因表达存在一些缺点,因此,本研究拟开发电致化学发光(ECL)技术来检测和量化EZH2 mRNA的表达量。在本研究中,用生物素和三(2,2-联吡啶)钌(II)(TBR)分别标记在PCR引物的5’末端上,用作扩增靶基因,扩增产物用ECL系统进行检测。我们用癌细胞作为模型分析了该方法的有效性和灵敏度,并且将其应用于25例乳腺癌的临床样本中EZH2基因表达量的检测。检测结果表明,EZH2基因在肿瘤细胞系中过量表达,而在正常血细胞中则低表达。最重要的是,在25例临床乳腺癌样品中发现10例样品(40%)的EZH2 mRNA过度表达。此方法提供了一种新的工具来评估EZH2基因在乳腺癌中的表达水平,且有可能成为一种快速、简便和灵敏的乳腺癌检测和诊断方法。     

Fourier transform infrared spectroscopic signature of blood plasma in the progression of breast cancer with simultaneous metastasis to lungs

Despite advanced diagnostic techniques used for detecting cancer, this disease still remains a leading cause of death in the developed world. What is more, the greatest danger for patients is not related with growing of tumor but rather with metastasis of cancer cells to the distant organs. In this study, Fourier transform infrared (FTIR) spectroscopy was used to track chemical changes in blood plasma to find spectral markers of metastatic breast cancer during the disease progression. Plasma samples were taken 1‐5 weeks after orthotropic inoculation of 4T1 metastatic breast cancer cells to mice. The earliest changes detected by FTIR spectroscopy in plasma were correlated with unsaturation of phospholipids and secondary structures of proteins that appeared 2 and 3 weeks, respectively, after 4T1 cells inoculation (micrometastatic phase). Significant alternations in the content and structure of lipids and carbohydrates were identified in plasma at the later stages (macrometastatic phase). When large primary tumors in breast and macrometastases in lung were developed, all bands in FTIR spectra significantly differed from those at earlier phases of the cancer progression. In conclusion, we showed that each phase of the breast cancer progression and its pulmonary metastasis can be characterized by a specific panel of spectral markers.     

Audit of 6 years' experience of breast fine needle aspiration (FNA) cytology using the cytospin method; improvement through multidisciplinary clinical audit

Audit of 6 years' experience of breast fine needle aspiration (FNA) cytology using the cytospin method; improvement through multidisciplinary clinical audit
A breast FNA cytology service for palpable breast lumps was commenced in 1989 using the cytospin method. Over the following 6 years 2314 aspirates were received. The results were audited in detail in 1990, 1991/1992 and 1994. Multidisciplinary clinical audit meetings followed each audit cycle. Practice change was agreed after each audit. Each audit cycle was followed by demonstrable improvement in the complete sensitivity of the technique, being respectively 79%, 88% and 96%. The cytospin method is a viable alternative to the conventional smear method.     

Effects of lymphocytes and fibroblasts on the growth of human mammary carcinoma cells studied in short-term primary cultures

Summary Breast carcinomas commonly contain varying amounts of fibrous stroma and infiltrates of lymphoid cells. Dickson and Lippman (Endocrine Rev., 8,29, 1987) have proposed a model of growth regulation in breast cancer involving interactions between stroma and carcinoma cells. This model is based on results obtained with established cell lines. In an effort to bring experimentation closer to the clinical situation we have used short-term primary cultures from human breast cancer in co-cultures with lymphocytes and fibroblasts. Cultures were established in a chemically defined serum-free medium (CDM3). Cell types were characterized on the basis of live morphology and expression of vimentin and keratin 18. A semi-quantitative system was developed for measuring growth of epithelial cells, thus defining two indices: maximal growth index (GI-max) and growth rate (GR). Moderate-to-good growth was obtained from 34 out of 46 carcinoma samples (74%) and 30 out of 38 parallel samples of non-cancerous tissue (79%). Success in culture was negatively correlated with the amount of hard stroma but unrelated to age of patient or clinical status. Malignant epithelium was clearly identified in 12 out of 34 (35%) carcinoma samples. For the evaluation of responses of epithelial cells in co-cultures, the cultures from each sample were ranked according to GI-max. From 20 co-culture experiments using carcinoma samples, the following results were obtained: the highest GI-max was found in 11 of the co-cultures with lymphocytes; in six of the co-cultures with fibroblasts; in one case in the control culture without partner cells; and in two experiments there was no difference between controls and co-cultures. The corresponding values for non-cancerous samples were: 5 out of 17, 2/17, 2/17, and 8/17. Control experiments performed without partner cells confirmed that these differences in GI-max between cultures were beyond random variations. Four samples displayed particularly vigorous responses to lymphocytes, and two samples responded extensively to fibroblasts. In four of these six samples cancer cells proliferated. We conclude that it is feasible to use primary cultures of breast carcinomas for experimentation. Fibroblasts did not have very marked effects on epithelial cell growth, but, contrary to expectation, there was a clear tendency for lymphocytes to stimulate growth.