Efficacy and patient satisfaction of breast conserving therapy for central breast cancer by the B technique.

PURPOSE: To evaluate the oncologic safety and cosmetic results after breast cancer surgery for central breast cancer by the B technique. METHODS: Seventy women with operable breast cancer located in the central portion of the breast that had received resection surgery with the B technique were recruited. The primary outcome was the oncological safety, quantified as rate of positive resection margins and the cosmetic outcome evaluated by postsurgical self-assessment of the cosmetic outcome via questionnaire. The median follow-up period was 61.4 months (range 7.9-142.6 months). RESULTS: With one exception all patients had T1-2 tumors less than 5 cm in diameter. Most patients had invasive ductal breast cancers (57.1 %), followed by ductal carcinoma-in situ (27.1 %) and invasive lobular breast cancers (8.6 %). The incidence of positive resection margins was 17.1 %. No local tumor recurrence occurred during follow-up; one patient had distant metastases. In total, 80 % of the patients reported that the cosmetic results met or exceeded their expectations. CONCLUSIONS: The B technique is a safe breast conservation surgery for the excision of tumors located in the central portion of the breast and yields a high rate of satisfactory cosmetic results.

Accidental intoxication with Veratrum album.

A 49-year-old man consumed two glasses (approximately 2 x 20 mL) of a beverage containing yellow gentian (Gentiana lutea). Shortly after ingestion, he developed nausea, vomiting, and oral paraesthesia. On admission to the hospital he suffered from severe bradycardia (35 beats/min) and hypotension (50/30 mm Hg), and he was treated with activated charcoal, antiemetics (metoclopramide, ondansetron), atropine, and intravenous electrolytic solution. The initial suspicion of Veratrum poisoning could be confirmed by identifying protoveratrines A (ProA) and protoveratrine B (ProB) in a sample from the beverage as well as in the patients serum by liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS-MS). The yellow-colored beverage contained 25% ethanol (by headspace gas chromatography), 20.4 mg/L ProA, and 13.7 mg/L ProB. The serum concentration of ProA was 1162 ng/L and ProB was 402 ng/L. Veratridine, cevadine, and jervine were not detected, neither in the beverage nor in the serum sample. The lower limits of quantitation for all compounds is 10 microg/L (S/N > 10, beverage) and 100 ng/L (S/N > 10, serum). After treatment, the patient completely recovered from the symptoms within 24 h and was discharged from the hospital. The analytical method described was developed for the simultaneous identification and quantitation of five Veratrum alkaloids. The method is based on a liquid-liquid extraction followed by LC-MS-MS analysis. The time needed for analysis was 6 min.

The clinical value of preoperative wire localization of breast lesions by magnetic resonance imaging--a multicenter study.

To evaluate the clinical value of MRI guided preoperative wire localization of clinically and mammographically occult lesions of the breast. In a multicenter study, we evaluated 132 preoperative MRI guided localizations. Median lesion size evaluated by MRI prior to wire localization was 9mm. MRI guided localization was successfully performed in 96.2% of cases. Median wire deviation from the lesion was 0 (0-10) mm. Moderate bleeding with no further treatment required occurred in three patients. We conclude that MRI guided preoperative wire localization is a safe and accurate procedure in cases of clinically and mammographically occult lesions of the breast.

[Preoperative chemotherapy in primary operable breast cancer with a dose-dense combination of doxorubicin and docetaxel (ADoc) - Experience of the GEPARDO-GABG study group]. / Präoperative Chemotherapie operabler primärer Mammakarzinome mit einer dosisdichten Kombination von Adriamycin und Docetaxel (ADoc) - Die Erfahrungen der GEPARDO-GABG-Studiengruppe -

OBJECTIVE: The German Adjuvant Breast Cancer Study Group (GABG) conducts trials of preoperative chemotherapy in patients with primary breast cancer using a combination of doxorubicin and docetaxel (ADoc). - PATIENTS AND METHODS: We conducted a parallel-grouped phase IIa-study with 42 patients with a conventionally dosed and a dose-dense ADoc-schedule (4 cycles of Doxorubicin 50 mg/m(2), Docetaxel 75 mg/m(2) i. v. day 1, q day 15 or 22; G-CSF day 3-15 only for the dose-dense schedule) and a randomized phase IIb-study (GEPARDO-Study) with 250 patients with ADoc +/- Tamoxifen. Biological factors were determined immunohistochemically on 197 core biopsies before treatment. A comparison to a sequential AC-Doc regimen including 913 patients has been completed recently. - RESULTS: ADoc can be applicated on schedule in 93 % of all patients. The dose-dense regimen shows a tendency to more toxicity but also to more efficacy. The rate of complete pathological remissions (pCR) was 9.7 %. No difference was found between chemo- and chemoendocrine treatment. Clinically negative lymphnodes and a negative estrogen receptor status is predictive for a higher pCR-rate. To date no differences in toxicity could be found between ADoc and AC-Doc. - CONCLUSIONS: The dose-dense ADoc regimen is well tolerated and highly effective as preoperative therapy of breast cancer.

Prognostic significance of local recurrence in breast cancer after postmastectomy radiotherapy.

PURPOSE: We have retrospectively analyzed the impact of local recurrence in patients with adjuvant radiation therapy after mastectomy for breast cancer. PATIENTS AND METHODS: From January 1985 through December 1993, 959 patients were irradiated after mastectomy for breast cancer. The age ranged from 34 to 79 years, the median follow-up was 3.1 years (range: 0.3-12.2 years). 368 (38%) were pre- and 591 (62%) postmenopausal. 35% had T3-4 tumors, 62% had axillary lymph node involvement, and 66% received additional systemic hormonal and/or cytotoxic therapy. Postmastectomy radiotherapy was administered in case of positive axillary nodes and in high-risk pN0-patients. The chest wall and lymphatics (axilla, parasternal and supraclavicular nodes) were irradiated with an anterior photon field with 50 Gy and the chest wall with an electron field with 44 Gy in 2-Gy fractions. RESULTS: The overall survival was 70.5% after 5 and 59.8% after 10 years. 53 patients (5.5%) developed a locoregional recurrence 2-96 months after treatment (median 26 months). The local control rate was 92.7% after 5 and 86.4% after 10 years. Axillary lymph node involvement was the most important and (in a multivariate analysis the only) risk factor for local recurrence (p = 0.0001). Patients with local control had a significantly better 10-year distant-disease-free survival and overall survival as compared to patients with local recurrence (44.5% vs 15.4%, p = 0.002 and 62.1% vs 34.8%, p = 0.004). Local recurrence increased the risk of death by a factor of 1.7 and in a Cox regression model, axillary lymph node status, T-category and local recurrence were significant prognostic factors for overall survival. In patients with local recurrence, the initial axillary lymph node status was the most important prognostic factor for survival after local recurrence. The 3-year survival after local relapse was 86% for patients with pN0 status vs 27% in with positive axillary nodes (p = 0.025). CONCLUSIONS: Local recurrence after treatment of breast cancer with mastectomy + radiotherapy +/- systemic therapy is associated with a significantly higher risk of distant metastases and death. In this analysis, local recurrence was a strong and, besides lymph node status and T category, an independent risk factor for survival. Minimizing the risk of local recurrence is therefore an essential goal of a curative treatment concept.

[New knowledge regarding tumor cell inactivation and histological evaluation after radiofrequency therapy. Single case observation and in vitro proof of a new hypothesis]. / Neue Erkenntnisse über Tumorzellinaktivierung und histologische Beurteilbarkeit nach Radiofrequenztherapie. Einzelfallbeobachtung und In-vitro-Nachweis einer neuen Hypothese.

In a study, a radiofrequency (RF) treatment was performed on a patient with a small breast cancer after vacuum biopsy. As usual in cases with a malignant diagnosis, surgical excision and axillary dissection followed. Histopathology revealed some residual tumor in the margin of the cavity. It could not be distinguished from vital tumor on the hematotoxylin eosin (HE) stain. Based on the correlation of MRI and histopathology after subsequent surgical excision, we did, however, presume that the residual was contained within the zone of inactivation. Thus the hypothesis arose that, if too high temperatures can be avoided, it might be possible to inactivate tumor cells without significantly impairing histopathologic assessment. This hypothesis was supported by the following in vitro experiment performed on a fresh specimen: An RF treatment was performed using temperatures up to 70 degrees C only. Half of the specimen underwent HE-staining, the other half vitality testing. The results indicate that if a given temperature range is strictly observed it appears possible to inactivate tissue before tissue sampling, since histopathologic diagnosis will not be impaired. Further technologic improvements may eventually allow to develop a pre-treatment method which might permit to avoid potential hematogenous tumor spread during subsequent biopsy.

Loss of transposase-DNA interaction may underlie the divergence of mariner family transposable elements and the ability of more than one mariner to occupy the same genome.

Mariners are a large family of eukaryotic DNA-mediated transposable elements that move via a cut-and-paste mechanism. Several features of the evolutionary history of mariners are unusual. First, they appear to undergo horizontal transfer commonly between species on an evolutionary timescale. They can do this because they are able to transpose using only their own self-encoded transposase and not host-specific factors. One consequence of this phenomenon is that more than one kind of mariner can be present in the same genome. We hypothesized that two mariners occupying the same genome would not interact. We tested the limits of mariner interactions using an in vitro transposition system, purified mariner transposases, and DNAse I footprinting. Only mariner elements that were very closely related to each other (ca. 84% identity) cross-mobilized, and then inefficiently. Because of the dramatic suppression of transposition between closely related elements, we propose that to isolate elements functionally, only minor changes might be necessary between elements, in both inverted terminal repeat and amino acid sequence. We further propose a mechanism to explain mariner diversification based on this phenomenon.

Gene discovery and gene function assignment in filamentous fungi.

Filamentous fungi are a large group of diverse and economically important microorganisms. Large-scale gene disruption strategies developed in budding yeast are not applicable to these organisms because of their larger genomes and lower rate of targeted integration (TI) during transformation. We developed transposon-arrayed gene knockouts (TAGKO) to discover genes and simultaneously create gene disruption cassettes for subsequent transformation and mutant analysis. Transposons carrying a bacterial and fungal drug resistance marker are used to mutagenize individual cosmids or entire libraries in vitro. Cosmids are annotated by DNA sequence analysis at the transposon insertion sites, and cosmid inserts are liberated to direct insertional mutagenesis events in the genome. Based on saturation analysis of a cosmid insert and insertions in a fungal cosmid library, we show that TAGKO can be used to rapidly identify and mutate genes. We further show that insertions can create alterations in gene expression, and we have used this approach to investigate an amino acid oxidation pathway in two important fungal phytopathogens.

Mutagenesis of Neisseria meningitidis by in vitro transposition of Himar1 mariner.

Now that the meningococcal genome sequence has been completed, the lack of a suitable method for saturation mutagenesis remains a major obstacle to the unraveling of the pathogenic propensity of Neisseria meningitidis. Here, we demonstrate that in vitro Himar1 mariner transposition on chromosomal or PCR-amplified meningococcal DNA, which is subsequently reintroduced into N. meningitidis by natural transformation, is an extremely efficient mutagenesis method. Southern blot analysis, sequencing the Himar1 insertion point in numerous transposition mutants, and a limited screening of the mutant libraries for clones impaired in maltose catabolism confirmed that Himar1 transposed randomly in N. meningitidis. Taken together, these data demonstrate that Himar1 in vitro transposition can lead to the exhaustive mutagenesis of N. meningitidis, allowing for the first time a genomic-scale mutational analysis of this important human pathogen.

In vivo transposon mutagenesis of the methanogenic archaeon Methanosarcina acetivorans C2A using a modified version of the insect mariner-family transposable element Himar1.

We present here a method for in vivo transposon mutagenesis of a methanogenic archaeon, Methanosarcina acetivorans C2A, which because of its independence from host-specific factors may have broad application among many microorganisms. Because there are no known Methanosarcina transposons we modified the mariner transposable element Himar1, originally found in the insect Hematobia irritans, to allow its use in this organism. This element was chosen because, like other mariner elements, its transposition is independent of host factors, requiring only its cognate transposase. Modified mini-Himar1 elements were constructed that carry selectable markers that are functional in Methanosarcina species and that express the Himar1 transposase from known Methanosarcina promoters. These mini-mariner elements transpose at high frequency in M. acetivorans to random sites in the genome. The presence of an Escherichia coli selectable marker and plasmid origin of replication within the mini-mariner elements allows facile cloning of these transposon insertions to identify the mutated gene. In preliminary experiments, we have isolated numerous mini-mariner-induced M. acetivorans mutants, including ones with insertions that confer resistance to toxic analogs and in genes that encode proteins involved in heat shock, nitrogen fixation, and cell-wall structures.

Direct measurement of sound velocity in various specimens of breast tissue.

RATIONALE AND OBJECTIVES: Previous studies using ultrasound CT or clinical amplitude/velocity reconstruction imaging ultrasound may indicate that cancers differ from normal breast tissue by increased sound velocity. However, only limited experience with direct measurements of sound velocity exists. This study aimed to investigate sound velocity measured directly in a variety of breast specimens. METHODS: Sound velocity was measured directly by forceps in fresh breast specimens chosen to contain one type of tissue only. Eighty specimens (31 cancers, 18 benign changes of glandular/fibrous tissue, 22 fatty tissues, 5 fibroadenomas, 2 compound tissues, 1 phylloides tumor, and 1 inflammation) were analyzed. RESULTS: Ultrasound velocities in carcinoma, benign changes, fibroadenoma, inflammation, and the phylloides tumor were very similar, with almost complete overlap. In contrary, the ultrasound velocity of fatty tissue was significantly lower. Compound tissues containing fat had an intermediate sound velocity. CONCLUSIONS: Sound velocity may add complementary information to echogenicity (B-scan). Because fat lobules exhibit low ultrasound velocity and carcinomas do not, a locally exact combination of ultrasound velocity information and reflexivity information should allow improved breast cancer detection by ultrasound.

In situ and minimally invasive breast cancer: morphologic and kinetic features on contrast-enhanced MR imaging.

PURPOSE: This retrospective study was undertaken to investigate the morphologic and dynamic features of in situ and minimally invasive breast cancer on contrast-enhanced (c.-e.) MR imaging and to examine possible associations to pathology features. MATERIAL AND METHODS: A total of 71 patients underwent MR imaging. T1-weighted FLASH-3D images were obtained before and after intravenous administration of Gd-DTPA. Histopathologic analysis of 78 lesions revealed ductal carcinoma in situ (DCIS) n = 50 and DCIS with microinvasion n = 28. MR features were correlated with histopathologic findings. RESULTS: Enhancement in DCIS was focal (73%), diffuse (10%) or ductal (17%). No enhancement occurred in two cases (4%). In 65% enhancement speed was classified as delayed. There was a tendency toward a more ill-defined (83 vs. 43%) enhancement pattern in high grade DCIS and a more ductal (29 vs. 12%) and faster (50 vs. 29%) enhancement in comedo type DCIS. However, significant differences in the enhancement behaviour could neither be demonstrated between high grade and non high grade DCIS nor between comedo and non comedo type DCIS. No significant differences were noted between pure and microinvasive DCIS. CONCLUSION: In this retrospective analysis the majority (96%) of DCIS lesions show contrast enhancement. However, in only about 50% of DCIS the criteria of a so-called 'typical' enhancement behaviour was fulfilled, that means strong, early, focal ill-circumscribed or ductal. Enhancement that follows a duct is often associated with malignancy, however this feature was only present in 17% of the cases. c.-e. MR imaging allowed the detection of 25 additional foci of DCIS. Therefore malignant in situ lesions can be present with atypical enhancement, and should be taken into consideration in high-risk patients in particular.

MR-guided percutaneous excisional and incisional biopsy of breast lesions.

The aim of this study was the realisation and clinical application of MR-guided vacuum biopsy for percutaneous excisional and incisional biopsy of enhancing breast lesions. A breast biopsy system and procedure have been developed which allow precise and safe access to breast lesions in any location and use of vacuum biopsy (VB) under MR guidance. Fifty-one patients with 55 MR-detected lesions were examined. Verification of these diagnoses included re-excision histology of all 14 malignancies and for benign lesions retrospective correlation of histology and imaging, assessment of complete or partial removal of the enhancing area directly after VB (40 of 40 lesions) and follow-up MRI (33 of 40 lesions), which in contrast to conventional needle biopsy can be used as proof of representative removal. Fifty-four of 55 procedures (including 15 lesions

Expression of lacunin, a large multidomain extracellular matrix protein, accompanies morphogenesis of epithelial monolayers in Manduca sexta.

Morphogenesis is a complex process operating at several levels of organization--organism, tissues, cells, and molecules. Complex interactions occur between and within these levels. Many of the molecules that mediate these interactions are predictably turning out to be large multidomain proteins. Here we describe one such novel protein associated with remodeling of epithelial monolayers in embryos and developing wings of the moth Manduca sexta. On the basis of its sequence and its expression pattern along lacunae of developing wings, we propose the name lacunin for this extracellular matrix protein that contains nine different types of domains, most of which are present in multiple copies. These include domains of various types: Kunitz proteinase inhibitors, thrombospondin type I, immunoglobulin-like, and several newly defined domains of unknown function (PAL, PLAC, and lagrin domains). This rich patchwork of distinct domains probably exerts multiple effects on a variety of cell behaviors associated with the complex phenomenon of epithelial morphogenesis.

Hyperactive transposase mutants of the Himar1 mariner transposon.

Mariner-family transposable elements are active in a wide variety of organisms and are becoming increasingly important genetic tools in species lacking sophisticated genetics. The Himar1 element, isolated from the horn fly, Haematobia irritans, is active in Escherichia coli when expressed appropriately. We used this fact to devise a genetic screen for hyperactive mutants of Himar1 transposase that enhance overall transposition from approximately 4- to 50-fold as measured in an E. coli assay. Purified mutant transposases retain their hyperactivity, although to a lesser degree, in an in vitro transposition assay. Mutants like those described herein should enable sophisticated analysis of the biochemistry of mariner transposition and should improve the use of these elements as genetic tools, both in vivo and in vitro.

In vivo transposition of mariner-based elements in enteric bacteria and mycobacteria.

mariner family transposons are widespread among eukaryotic organisms. These transposons are apparently horizontally transmitted among diverse eukaryotes and can also transpose in vitro in the absence of added cofactors. Here we show that transposons derived from the mariner element Himar1 can efficiently transpose in bacteria in vivo. We have developed simple transposition systems by using minitransposons, made up of short inverted repeats flanking antibiotic resistance markers. These elements can efficiently transpose after expression of transposase from an appropriate bacterial promoter. We found that transposition of mariner-based elements in Escherichia coli produces diverse insertion mutations in either a targeted plasmid or a chromosomal gene. With Himar1-derived transposons we were able to isolate phage-resistant mutants of both E. coli and Mycobacterium smegmatis. mariner-based transposons will provide valuable tools for mutagenesis and genetic manipulation of bacteria that currently lack well developed genetic systems.

[Direct in-vitro measurement of ultrasound velocity in carcinomas, mastopathic tissue, fatty tissue and fibroadenomas of the female breast]. / Direkte In-vitro-Messung der Ultraschallgeschwindigkeit in Karzinomen, mastopathischem Gewebe, Fettgewebe und Fibroadenomen der weiblichen Brustdrüse.

PURPOSE: The study aimed to investigate ultrasound velocity (SV) in carcinomas, fibrocystic changes, fibroadenomas and fatty tissue of the female breast by means of direct in-vitro measurements. We intended to test whether or not differences in SV exist between the various types of tissue and whether the SV is a useful criterion to differentiate the different tissues. METHOD: SV was measured by comparing transmission time of the ultrasound beam through the specimen and through water. Altogether 40 specimens (12 cancer, 14 fibrocystic changes = FCD, 10 fatty tissues, 3 fibroadenomas, and 1 mixed tissue) were analysed. RESULTS: Velocity differed significantly between fat (1478.5 +/- 6.5 m/s) and tumor (1523.1 +/- 5.9 m/s) (p approximately 10(-11)) and between fat and FCD (1526.0 +/- 9.0 m/s) (p approximately 10(-12)). No significant differences and much overlap were seen between the ultrasound velocities of tumors and FCD. Ultrasound velocity in fibroadenomas (1533.2 +/- 3.8 m/s) was comparable with that in carcinomas and FCD. CONCLUSIONS: We conclude that ultrasound velocity may add complementary information to echogenicity (B-scan). Thus, a locally exact correlation of echogenicity and sound velocity might allow for an improved tissue characterization.

The Himar1 mariner transposase cloned in a recombinant adenovirus vector is functional in mammalian cells.

Mariner transposons belong to the mariner /Tc1 superfamily of class II, DNA-mediated elements. One of these transposons, Himar1 , isolated from the horn fly, is independent of host-specific factors that would limit transfer between different species, making it an ideal candidate for gene transfer technology development. To determine the activity of Himar1 transposase in mammalian cells, we introduced the Himar1 transposase gene into an adenovirus (Ad) vector under control of the phage T7 RNA polymerase promoter. Mammalian cells infected with the Ad vector carrying the Himar1 gene efficiently expressed the Himar1 transposase in the presence of T7 polymerase. In in vitro inter-plasmid transposition reactions, Himar1 transposase expressed by the Ad vector mediated precise cut-and-paste transposition and resulted in a characteristic duplication of TA at the integration site of the target plasmid. Further studies showed that this transposase was capable of catalyzing transposition between twoplasmids co-transfected into 293T7pol cells, which express T7 RNA polymerase. Combining the integration capability of mariner transposons with the transduction efficiency of Ad vectors is expected to provide a powerful tool for introducing transgenes into the host chromosome.

[Preoperative axilla sonography in breast tumor suspected of malignancy--a diagnostic advantage?]. / Präoperative Axillasonographie beim malignitätsverdächtigen Mammatumor--ein diagnostischer Gewinn?

PURPOSE: To assess the value of preoperative axillary sonography possibly malignant for breast tumours. METHOD: We performed preoperative axillary sonography on 89 patients with suspicious breast tumours. In 78 cases, among which there were 74 invasive carcinomas, the surgery that followed included an axillary lymph node dissection and a comparison with the histology was possible. RESULTS: The sonographic detection of axillary lymph node metastases has a sensitivity of 90% and a specificity of 91.7% in relation to all tumour stages and a sensitivity of 100% and specificity of 89.6% in relation to T1 tumours. In our analysis, therefore, it is vastly superior to a clinical examination of the axilla. CONCLUSION: Ultrasonography is an accurate imaging method for the diagnosis of axillary lymph node metastases. Broadening this analysis to include more patients will validate this conclusion. The results of our preliminary study suggest that this diagnostic method could help reduce unnecessary radical surgery in the treatment of breast cancer.