Whole genome expression analysis for biologic rational pathway modeling: application in cancer prognosis and therapy prediction.

Using semi-quantitative microarray technology, almost every one of the approximately 30 000 human genes can be analyzed simultaneously with a low rate of false-positives, a high specificity, and a high quantification accuracy. This is supported by data from comparative studies of microarrays and reverse-transcription PCR for established cancer genes including those for epidermal growth factor receptor (EGFR), human epidermal growth factor receptor-2 (HER2/ERBB2), estrogen receptor (ESR1), progesterone receptor (PGR), urokinase-type plasminogen activator (PLAU), and plasminogen activator inhibitor-1 (SERPINE1). As such, semi-quantitative expression data provide an almost completely comprehensive background of biological knowledge that can be applied to cancer diagnostics. In clinical terms, expression profiling may be able to provide significant information regarding (i) the identification of high-risk patients requiring aggressive chemotherapy; (ii) the pathway control of therapy predictive parameters (e.g. ESR1 and HER2); (iii) the discovery of targets for biologically rational therapeutics (e.g. capecitabine and trastuzumab); (iv) additional support for decisions about switching therapy; (v) target discovery; and (vi) the prediction of the course of new therapies in clinical trials. In conclusion, whole genome expression analysis might be able to determine important genes related to cancer progression and adjuvant chemotherapy resistance, especially in the context of new approaches involving primary systemic chemotherapy. In this review, we will survey the current progress in whole genome expression analyses for cancer prognosis and prediction. Special emphasis is given to the approach of combining biostatistical analysis of expression data with knowledge of biochemical and genetic pathways.

Video-thoracoscopy and staged management of preoperative empyema in lung cancer.

The incidence of pleural empyema as a primary finding in lung cancer patients is low (0.1 to 0.3 %) and the management of those patients deemed operable consists of either infection control prior to resection, or thoracotomy and resection before infection control. We successfully resolved (in one patient) and alleviated (in a second patient) empyema-related systemic infection by video-assisted thoracic surgery (VATS), and were thus able to resect their lung tumours 14 and 21 days later, respectively.

Effects of a bradycardic agent on postischemic cardiac recovery in rabbits.

Decreasing heart rate might be beneficial for improvement of myocardial energetics and could reduce the severity of myocardial ischemia. We examined the contribution of heart rate reduction by cilobradine (DK-AH 269), a direct sinus node inhibitor, on left ventricular function and peripheral vasomotion in anesthetized rabbits with experimental myocardial infarction. The rabbits were randomized to receive either placebo (n=10) or cilobradine (n=7). Cilobradine decreased significantly heart rate from 163 +/- 33 to 131 +/- 13 bpm, p< 0.05, without any inotopic or vascular effects. After 60 min coronary occlusion and 30 min reperfusion, both systolic and diastolic ventricular function were more reduced in the cilobradine group; i.e. maximal left ventricular pressure significantly decreased to 62 +/- 11 mmHg, p < 0.05 (placebo: 77 +/- 9 mmHg); dP/dt(min) significantly decreased to -904 +/- 247 mmHg, p < 0.05 (placebo: -1106 +/- 242 mmHg). However, infarct size in the cilobradine group was significantly smaller compared with the placebo group. In conclusion, cilobradine reduced heart rate without any negative inotropic effect and reduced infarct size. On that account, this bradycardic agent might open a promising therapeutical avenue to treat postischemic dysfunction.

[Remission rate of mediastinal lymph nodes after multimodal therapy of lung cancer--is it a prognostic factor?]. / Remission mediastinaler Lymphknoten nach multimodaler Therapie des Bronchialkarzinoms--ein Prognosefaktor?

OBJECTIVE: General acceptance in the oncologic community has been gained for combined modality treatment of non-small cell lung (NSCLC) cancer in locally advanced stage IIIA and IIIB disease. However, no optimal regimen has been established. This study (chemotherapy and radiochemotherapy followed by operation) assesses feasibility, response, resectability, and survival in patients with stage IIIA and IIIB lung cancer. Currently, only little data is available about the prognostic significance of tumor clearance of mediastinal nodes. Thus, an important aim of our study was to evaluate the prognostic significance of the extent of tumor reduction in mediastinal nodes by a neoadjuvant multimodality protocol. PATIENTS: In a phase II protocol, 26 patients underwent neoadjuvant radiochemotherapy. Subsequently, a radical lymphadenectomy was performed during surgery. The extent of tumor regression was determined according to the methodology initially described by Salzer-Kuntschik for osteosarcoma: Grade I: no vital tumor cells, grade II: some tumor cells, grade III: less than 10 % vital tumor cells, grade IV: 10-50 % vital tumor cells, grade V: more than 50 % vital tumor cells, grade VI: no effect of chemotherapy. RESULTS: Complete pathologic response was seen in 30.7 % of primary tumors, in 38.5 % of mediastinal lymph nodes, and in 23 % of corresponding specimens simultaneously. Median survival was 34.7 months for those patients with grade I, 12.6 months with grade II, and 8.9 months for patients showing a grade III/IV regression in mediastinal nodes. Response rate to neoadjuvant chemotherapy in mediastinal nodes proved to be the only statistically significant parameter for long-term survival: In cases with no vital tumor cells in the operation specimen, median survival was 34.7 months in comparison to those with vital cells showing a median survival of only 11.4 months (P = 0.01). CONCLUSION: Patients with locally advanced NSCLCs can enjoy long-term survival after multimodal therapy. However, the complications related to therapy are considerably. Especially, clearance of tumor cells from mediastinal lymph nodes is an important independent prognostic factor.

p53 Gene status in relation to ex vivo chemosensitivity of non-small cell lung cancer.

PURPOSE: About 40% of non-small cell lung cancer (NSCLC) patients are candidates for systemic chemotherapy, despite the fact that at diagnosis most NSCLC are usually chemoresistant both in vivo and ex vivo. It is important to develop sufficient methods of prediction of the response to chemotherapy and to find molecular markers that may prognose this response. Therefore, a study on the relationship of p53gene status to the ex vivo chemosensitivity of primary human NSCLC was performed. METHODS: Three drug combinations (carboplatin/etoposide, cyclophosphamide/etoposide/epirubicin, and paclitaxel/carboplatin) were tested in a modified ATP cell viability assay. A group of 28 cases of primary human NSCLC was assessed. RESULTS: Ex vivo chemosensitivity testing showed that tumors with p53 mutations were significantly more resistant to the cyclophosphamide/etoposide/epirubicin regimen than with normal p53 gene ( P = 0.012). However, no correlation was observed for two other treatment regimens. CONCLUSION: Mutations in the p53gene can lead to enhanced chemoresistance, confirming the hypothesis that the p53 gene may serve as a marker of tumor response to treatment in NSCLC. However, the data also illustrate that some additional factors might contribute to drug resistance of the examined tumors.

Early and late results after thymectomy in myasthenia gravis: a retrospective study [correction of analysis].

BACKGROUND: This study aims to evaluate the early and late outcome of patients treated by surgery for myasthenia gravis and the diagnostic value of the Besinger Score, which is based on a correlation of severity of symptoms with specific antibodies to acetylcholine receptors, in the follow-up investigation after surgical therapy. METHODS: Between June 1984 and April 1992 thoracotomy was performed in 51 myasthenia gravis cases at our department. The retrospective analysis considered patients with (n = 13) or without thymoma (n = 38). The Besinger score was used to describe the severity of disease preoperatively and up to 5 years postoperatively. RESULTS: The Besinger score fell continually post surgery. Changes in relative serum concentrations of antibodies were similar to the Besinger score. Five years after thymectomy complete remission was diagnosed in 40% of the patients. The required dosage of pyridostigmine had fallen by two thirds after 5 years. Patients with follicular hyperplasia had significantly higher remission rates than those with thymoma. CONCLUSIONS: Surgery for myasthenia gravis is successful. The Besinger score well quantifies the severity of the disease.

Pharmacologic heart rate reduction: effect of a novel, specific bradycardic agent on the heart.

Because heart rate (HR) is a major determinant of myocardial oxygen consumption (MVO2) a decrease in HR could prevent ischemia or reduce its consequences. We examined the effects of a novel bradycardic agent of the benzazepinone type, DK-AH 269 (DK), on ventricular function and perfusion in 12 isolated, blood-perfused rabbit hearts. HR was significantly reduced by 1mumol/L DK (160 +/- 28 vs. 124 +/- 23 min-1); diastole lengthened from 235 +/- 69 to 334 +/- 85 ms. Aortic flow tended to fall after DK (50.0 +/- 29.6 vs. 35.6 +/- 21.5 ml/min), but stroke volume remained unchanged (0.29 +/- 0.16 vs. 0.28 +/- 0.17 ml) following DK. Peak left-ventricular pressure (LVPmax) (106 +/- 29 vs. 92 +/- 35 mmHg) and dp/dtmax (1482 +/- 582 vs. 1247 +/- 644 mmHg/s) were decreased. dp/dtmin, as a measure of early relaxation, was also decreased (-1361 +/- 362 vs, -1125 +/- 488 mmHg/s), whereas the end-diastolic pressure (LVPed) was increased (20 +/- 12 vs. 25 +/- 17 mmHg). Coronary blood flow (CBF) per beat was not affected by DK: 0.07 +/- 0.02 vs. 0.07 +/- 0.02 ml. However, the coronary resistance increased with DK from 0.76 +/- 0.29 to 1.13 +/- 0.66 mmHg/(ml/min/100 g). The MVO2 was decreased (6.8 +/- 3.4 vs. 5.9 +/- 2.8 ml/min/100 g). The relation between subendocardial and subepicardial flow (colored microspheres) was unchanged after DK (1.12 +/- 0.22 vs. 1.13 +/- 0.16). Using electrical pacing to restore the control HR, dp/dtmax, LVPed, and MVO2 were nearly restored to predrug levels. In contrast, stroke volume, LVPmax, dp/dtmin and CBF per beat were less than control. In summary: DK effectively reduces heart rate and increases diastole. In parallel with the moderately reduced contractile function, MVO2 is reduced whereas CBF per beat is preserved. These results suggest that this novel bradycardic agent could be useful in treating unwanted tachycardia in the experimental setting, postoperative tachycardia in patients with heart disease or be useful even in treating coronary heart disease.

[Effect of a new bradycardic substance on the isolated rabbit heart]. / Effekt einer neuen bradykarden Substanz auf das isolierte Kaninchenherz.

Beside wall tension and contractility, heart rate is a major determinant of myocardial oxygen consumption. Therefore, a decrease in heart rate could prevent ischemia or reduce its consequences. We examined the effect of a new bradycardic agent of the benzazepinone-type (DK-AH 269) on eight isolated, saline-perfused rabbit hearts, bradycardia resulted from a specific blockade of i(f)-channels in sinus node cells. After control measurements (C), the substance was added in three increasing concentrations (D1: 10(-8) M, D2: 10(-7) M, D3: 10(-6) M). We observed a dose-dependent reduction in heart rate (C: 206 +/- 25, D1: 195 +/- 30, D2: 77 +/- 41, D3: 154 +/- 48/min). In the highest dosage, the duration of diastole was increased by 100%. To characterize systolic function, we measured stroke volume (SV), peak left ventricular pressure (LVPmax) and its first derivative (dP/dtmax). Aortic flow was slightly decreased whereas SV increased to 108% of control after initial reduction at the two lower dosages. LVPmax remained unchanged, and dP/dtmax was dose-dependently reduced to 91, 81, and 70% of control (C: 1885 +/- 376, D1: 1721 +/- 525, D2: 1526 +/- 504, D3: 1327 +/- 337 mm Hg/s); dP/dtmin as a measure of early relaxation was also reduced. The coronary flow per beat did not change compared with control in the presence of the two lower doses of DK-AH 269, but was significantly increased with the highest dose (C: 0.29 +/- 0.06, D1: 0.28 +/- 0.07, D2: 0.29 +/- 0.09, D3: 0.34 +/- 0.11 ml). The myocardial oxygen demand was dose-dependently decreased (C: 10.4 +/- 2.5, D1: 9.6 +/- 2.5, D2: 8.8 +/- 2.6, D3: 7.9 +/- 2.4 ml/min/100 g). The relation between subendocardial and subepicardial flow, assessed with colored microspheres, exhibited no changes in the presence of the highest dose of DK-AH 269 (C: 1.28 +/- 0.09, D3: 1.27 +/- 0.08). DK-AH 269 reduced heart rate in isolated rabbit hearts and increased the duration of diastole. Whereas systolic function was primarily left unchanged, coronary flow per beat and oxygen consumption were decreased. According to our results, this new bradycardic agent could be useful in treating coronary heart disease.

Cytokinetic investigations in human breast cancer by flow cytometrically recorded DNA/protein distributions.

This prospective study characterizes T1-T2 breast carcinomas (N = 114) and fibroadenomas (N = 16) by cell kinetic parameters derived from flow cytometrically recorded DNA/protein histograms. Ploidy level, cell cycle distribution and the number of cell subpopulations (SP) characterized by correlating DNA and protein values were assessed. The subpopulations were derived from the three-dimensional plot. The estrogen receptor (ER) status was determined biochemically (N = 61). Within the G1/0 cell peak 1-6 SP were evident in principle. Depending on the number of SP, two subsets were established: subset 1 with < or = 2 SP, subset 2 with > or = 3 SP. They differed significantly in proliferative activity expressed in the percentage of cells in the G2M phase. Subset 2 showed the higher activity. Analysis of subset distributions revealed that subset 1 prevails in favourable prognostic cases as ER positive cases (P < 0.03), lobular carcinomas (P < 0.01) and LN- cases (P < 0.03), whereas subset 2 prevails in the unfavourable counterparts. Analysis of variance showed that the main effect on proliferative activity indicated by G2M% is due to subpopulation composition rather than histologic type, nodal status or ER status (P < 0.01, P < 0.002, P < 0.05), not even due to ploidy level (P < 0.0001). The rationale for subset stratification may be cytogenetic variability connected with protein content heterogeneity accounting for kinetic SP.

[Primary seminoma. A rare mediastinal tumor]. / Das primäre Seminom. Ein seltener mediastinaler Tumor.

This is a report on three patients with primary mediastinal seminoma. Two patients had no symptoms, and one had had thoracic pain for the last few years. The preoperative diagnosis was thymoma in all cases, and in one patient the radiologist had suspected a seminoma. We removed the tumor after performing median sternotomy (n = 2) and anterolateral left thoracotomy (n = 1). The presence of a primary gonadal seminoma was excluded with a urological and ultrasound examination. All patients are still alive following adjuvant chemotherapy (n = 2; 120 and 8 months) and radiotherapy (n = 1; 84 months). Chemotherapy consisted of four cycles of cisplatin, etoposide and ifosfamide or combination therapy with cisplatin, bleomycin and velbe.

Relationship between 43 kDa epidermal growth factor-related clonogenic activity and clinical parameters for breast cancer.

The clonogenic activity of a previously purified 43 kDa EGF-related protein (16) was estimated in the urine of breast cancer patients. Increase of activity was statistically significant in lymph node-positive patients, in a group of patients with larger carcinomas, with accelerated tumor growth, in premenopausal patients and younger age and in estrogen receptor negative patients. In 31 patients the activity was estimated during polychemotherapy before surgery. Differences between the values at the begin and the end of treatment were compared between all groups by the Kruskal-Wallis test (p = 0.02). Patients with progressive disease showed increasing activities (mean values from 315 to 811) while in those with complete remission activity decreased (from 449 to 213). Partial- and non responders showed no change. In a long-term follow-up study with 25 patients the pre- and postoperative activities were estimated. In 17 of 18 patients who had no local recurrence or metastasis the activity declined after surgery, whereas in 6 of 7 patients who died, activity increased 2-5 fold prior to death. A life table analysis with a total of 101 patients revealed a trend to shorter survival in the group with higher activity (p = 0.042). These observations suggest a role for the EGF-like growth factor activity in the expression of the malignant phenotype and may have significance for breast cancer diagnosis and prognosis.

[Surgical and adjuvant therapy of breast cancer]. / Chirurgische und adjuvante Therapie des Mammakarzinoms.

This is a report on 512 patients operated on for primary breast cancer in 1988 and 1989 at the Central Institute of Cancer Research in Berlin-Buch. Most patients were postmenopausal (52%), 17.7% were aged 45 to 49 years. The average duration of the symptoms was 6.1 months. 45.4% of all the tumours were located in the upper outer quadrant, 47.3% in the right and 52.7% in the left breast. Clinical evaluation and mammography had a sensitivity of 89%, 93.7% respectively. Invasive ductal carcinomas were histologically proven in 319 cases, less frequently (n = 118) invasive lobular tumours. Predominant surgical procedures were the quadrantectomy with axillary dissection (33%) and the modified radical mastectomy according to Auchincloss-Madden (25.5%). In the case of medially/centrally located tumours and axillary lymph node involvement greater than or equal to 4 nodes independent of the tumour site we performed the IMN-biopsy in 129 patients. The majority of the premenopausal node positive patients received adjuvant chemotherapy (94.4%), postmenopausal node positive patients with positive ER received adjuvant tamoxifen. Postoperative irradiation indicated was applied in 25 patients.

A Mr 43,000 epidermal growth factor-related protein purified from the urine of breast cancer patients.

A tumor-associated epidermal growth factor (EGF)-like activity was detected in the urine of breast cancer patients by means of an EGF radioreceptor assay and an anchorage-independent growth assay. The clonogenic growth factor activity of pooled void volume eluate fractions from a Bio-Gel P-30 column was completely neutralized by an anti-human epidermal growth factor antiserum but not by an anti-transforming growth factor alpha antiserum. This activity was determined in the urine of 71 breast cancer patients. A statistically significant correlation was found between EGF-like clonogenic activity and axillary lymph node status, tumor size, stage of disease, and grade of differentiation of the primary tumor. The Bio-Gel P-30 void volume fraction was used to purify the EGF-related polypeptide to apparent homogeneity by subsequent binding to and elution from A431 cells followed by isoelectric focusing. A polypeptide of a pI of approximately 3.4 was identified to be related to EGF by neutralization and immunoprecipitation experiments with anti-human epidermal growth factor antisera. This polypeptide migrated as a single band of Mr 43,000 in sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

[Breast cancers in pregnancy and lactation]. / Mammakarzinome in Schwangerschaft und Laktationsperiode.

Four cases of rare coincidence of pregnancy and mammary carcinoma are described in this paper. Tumour growth was at an advanced stage and prognosis thus deteriorated due to delayed diagnosis. The therapeutic concept should be in keeping with the stage of tumour growth and should be formulated with due consideration of the patient's individual peculiarities.

Purification of a high molecular weight form of epidermal growth factor from urine of breast cancer patients.

A high molecular weight form of epidermal growth factor (EGF) was detected by means of an EGF radio-receptor assay and an anchorage-independent growth assay in the urine of breast cancer patients. Preliminary data indicate that the activity of this growth factor is associated with lymph node status and tumor size and that the activity becomes reduced after removal of the primary tumor. The EGF-related polypeptide was purified to homogeneity by a combination of Sephadex G-25 and Bio Gel P-30 chromatography followed by binding to, and elution from, EGF receptor rich A431 cells. Final purification was achieved after isoelectric focusing by following the biological activity of eluted polypeptides. A polypeptide of a pI of 3.4 was identified to carry EGF-like activity. This polypeptide migrated as a single band of 43 kDa in SDS-PAGE. Its biological activity was neutralized by a specific anti-hEGF-antibody indicating an immunological relationship with hEGF.

[Immunologic studies using the leukocyte adherence inhibition test in the diagnosis and postoperative follow-up of breast cancer]. / Immunologische Untersuchungen mit dem Leukozytenadhärenzinhibitionstest zur Diagnostik und postoperativen Verlaufsbeurteilung des Mammakarzinoms.

37 women with histologically proved carcinoma of the breast as well as 3 control groups (benign breast diseases, other carcinomas, benign surgical diseases) were investigated by means of the leucocyte adherence inhibition test. In the primary diagnostics of the carcinoma of the breast the specificity was 56%, the sensitivity 70%. Follow-up studies on the conditions of a district hospital over a period of 2 years showed a different reactivity which is suitable for the evaluation of recurrence and immunological tumour-host relations.