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Background Glioblastoma (GBM) is an aggressive brain tumor with a median survival of 14.6 months. We have no standard treatment for relapse and current treatment options have limited effect. Novel treatments are necessary to improve survival and quality of life. Material and Methods We present data from; a phase II open label, two-armed clinical trial studying nivolumab and bevacizumab in treatment of recurrent GBM, with progression after Stupp’s regime. Patients were included in two arms depending on the possibility of salvage neurosurgical resection. All patients had biopsies for genome sequencing at primary tumor and recurrence. Both arms received nivolumab and bevacizumab administrated every second week and the surgical arm also received neoadjuvant nivolumab 7 days prior to surgery. Fresh tissue samples were collected for tumor digest, TILs (tumor infiltrating lymphocytes) for phenotype exploration and intracellular staining to test reactivity. Patients were treated until progression, death, or intolerable side effects. Toxicity screens were reported, and follow-up ended in Marts 2022. Results Forty-four patients were included from November 2018 to January 2022; 20 in each arm (four screen-failures). Treatment was overall well tolerated. Median (m) age at inclusion was 57,5 years (arm A) and 50,5 years (arm B), and the groups had an even distribution. The surgical and non-surgical arm had an mPFS of 5.95 and 3.83 months respectively, while the mOS was 13.96 months and 6.77 months, respectively. Multivariate analysis was performed by variables such us steroid, MGMT, gender, age at diagnosis, resection extent and arm. Steroid at inclusion was a significant negative predictor of outcome (p = 0.0378). Controls from our GBM registry (N=140), which were treated with neurosurgical resection and then bevazicumab and irinotecan in recurrent setting had an mOS of 8.64 months (log-rank p=0.0181).Furthermore, reactive tumor infiltrating lymphocytes (TIL) were detected in four of the patients who presented with a longer mOS and mPFS of 16.75 months and 9.18 months, while the 16 patients without TIL reactivity had mOS and mPFS of 12.63 months and 5.13 months, respectively (not significant). Conclusion We found an increased mOS in patients treated with nivolumab and bevacizumab at recurrence, compared to our controls: 13.96 months and 8.64 months, respectively. Four patients with T-cell reactivity towards tumor cells showed an even longer mPFS and mOS. Though not significant, these results warrant further research evaluation in larger patient cohorts. We are currently investigating proteomics and sequencing data to identify predictive biomarkers.

We sought to assess the influence of the clinical introduction of new radiotherapy technologies on glioblastoma patients' outcomes.Newly diagnosed glioblastoma patients treated with 60 Gy and temozolomide (2005-2014) were analyzed. The patients' GTV and CTV were defined based on MR (n = 521) or FET-PET/MR (n = 190), and were treated using conformal radiotherapy (CRT, n = 159) or image-guided volumetric modulated arc therapy with hippocampal sparing (IG-VMAT, n = 362). Progression-free survival (PFS) was assessed using the McDonald criteria. Associations between clinical data, dosimetry data, treatment technology, for PFS and overall survival (OS) were explored.The PFS (7 months) and OS (15 months) were unaffected by CRT, IG-VMAT and FET-PET technology. Mean brain dose was correlated with tumor volume, and was lower for IG-VMAT vs. CRT (p 0.001). Larger mean brain dose was associated with inferior PFS (univariate/multivariate Cox models, p 0.001) and OS (univariate, p 0.001). Multivariate Cox models revealed association of larger mean brainstem dose (p 0.001), BTV (p = 0.045), steroid use at baseline (p = 0.003), age (p = 0.019) and MGMT status (p = 0.022) with lower OS.Introduction of hippocampal-sparing IG-VMAT technology appeared to be safe, and may have reduced toxicity and cognitive impairment. Larger mean brain dose was strongly associated with inferior PFS and OS.

BACKGROUND: Discrimination between recurrent glioblastoma and therapy-related changes with conventional magnetic resonance imaging (MRI) is challenging. There is increasing evidence for the clinical value of positron emission tomography (PET) with O-(2-18F-fluoroethyl)-L-tyrosine (FET) in addition to MRI in cases of ambiguous findings in the setting of posttreatment care. The diagnostic accuracy, however, varies among the published studies and is influenced by the composition of the population with different glioma subtypes/grades, lack of histological confirmation, and differences in data processing. Herein, we evaluated the diagnostic accuracy of FET PET scans in glioblastoma patients. MATERIAL AND METHODS: One hundred thirty-seven consecutive patients who had undergone 150 FET PET were reviewed retrospectively. Inclusion criteria were 1) histologically-proven glioblastoma; 2) previous surgery followed by oncological treatment consisting of standard radiochemotherapy or - if at second recurrence - chemotherapy; 3) unexplained constant or increasing contrast-enhancing (CE) lesions on T1, or non-CE lesions on T2 FLAIR MRI later than 6 months after radiotherapy, where the differentiation between disease recurrence or therapy-related changes was uncertain; 4) FET PET for supplementary evaluation; 5) a histological evaluation following surgery less than 3 months after FET PET, or MRI follow-up. FET PET scans were performed as 20-minute static PET/CT acquisitions and evaluated co-registered to T1 post-contrast MRI with measurement of maximum and mean tumor-to-brain ratios (TBR(max,) TBR(mean)). Receiver operating characteristics (ROC) analysis was used to determine the optimal threshold of FET parameters. The prognostic influence of FET parameters on overall survival (OS) was investigated with the Cox proportional hazards model. RESULTS: The median time interval from radiotherapy until the radiological progression was 13 months. One hundred twenty-six PET scans demonstrated FET uptake of varying intensity. Surgical interventions were performed following 88 PET scans, while 62 PET scans were evaluated by clinical or MRI follow-up, resulting in 131 glioblastoma recurrences and 19 therapy-related changes. ROC analysis yielded the thresholds of 2.0 for TBR(max) and 1.8 for TBR(mean) for differentiation between recurrent disease and therapy-related changes with the best performance of TBR(max) with a sensitivity of 96% and a specificity of 100% (p < 0.0001). Using this threshold, 145 of 150 PET scans of glioblastoma recurrence or therapy-related changes were accurately classified. Multivariate survival analysis showed that TBR(max) > 2.0 predicted independently a shorter OS in patients, who did not undergo subsequent therapy following PET (p = 0.002). CONCLUSION: FET PET is a powerful noninvasive tool to distinguish recurrent glioblastoma from therapy-related changes.

The correlation between glucose-6-phosphate dehydrogenase activity and different tumour characteristics was investigated in human breast cancer tissue. The enzyme activity was measured by a histochemical method and the oestrogen receptor content by a dextran-coated charcoal assay. The proliferative activity of the tumours correlated positively with the glucose-6-phosphate dehydrogenase activity. No correlation was found between enzyme activity and tumour type and menopausal status, age, TNM-class nor oestrogen receptor content. Based on this investigation it is concluded that the activity of glucose-6-phosphate dehydrogenase reflects the proliferative status of the tumour but not the hormone dependency of the tumour.

Objective: To evaluate a possible association of glioma or meningioma with use of cellular telephones, using a nationwide population-based case-control study of incident cases of meningioma and glioma. Methods: The authors ascertained all incident cases of glioma and meningioma diagnosed in Denmark between September 1, 2000, and August 31, 2002. They enrolled 252 persons with glioma and 175 persons with meningioma aged 20 to 69. The authors also enrolled 822 randomly sampled, population-based controls matched for age and sex. Information was obtained from personal interviews, medical records containing diagnoses, and the results of radiologic examinations. For a small number of cases and controls, the authors obtained the numbers of incoming and outgoing calls. They evaluated the memory of the respondents with the Mini-Mental State Examination and obtained data on socioeconomic factors from Statistics Denmark. Results: There were no material socioeconomic differences between cases and controls or participants and non-participants. Use of cellular telephone was associated with a low risk for high-grade glioma (OR, 0.58; 95% CI, 0.37 to 0.90). The risk estimates were closer to unity for low-grade glioma (1.08; 0.58 to 2.00) and meningioma (1.00; 0.54 to 1.28). Conclusion: The results do not support an association between use of cellular telephones and risk for glioma or meningioma.

Formation of metastasis is a multistep process involving attachment to the basement membrane, local proteolysis and migration into surrounding tissues, lymph or bloodstream. In the present study, we have analysed the correlation between in vitro invasion and presence of the epidermal growth factor receptor (EGFR) in a panel of 21 small-cell lung cancer (SCLC) cell lines. We have previously reported that ten of these cell lines expressed EGFR protein detected by radioreceptor and affinity labelling assays. In 11 small-cell lung cancer (SCLC) cell lines, EGFR mRNA was detected by Northern blot analysis. In vitro invasion in a Boyden chamber assay was found in all EGFR-positive cell lines, whereas no invasion was detected in the EGFR-negative cell lines. Quantification of the in vitro invasion in 12 selected SCLC cell lines demonstrated that, in the EGFR-positive cell lines, between 5% and 16% of the cells added to the upper chamber were able to traverse the Matrigel membrane. Expression of several matrix metalloproteases (MMP), of tissue inhibitor of MMP (TIMP) and of cathepsin B was evaluated by immunoprecipitation, Western blot analysis and reverse transcriptase polymerase chain reaction (RT-PCR). However, in vitro invasive SCLC cell lines could not be distinguished from non-invasive cell lines based on the expression pattern of these molecules. In six SCLC cell lines, in vitro invasion was also determined in the presence of the EGFR-neutralizing monoclonal antibody mAb528. The addition of this antibody resulted in a significant reduction of the in vitro invasion in three selected EGFR-positive cell lines. Our results show that only EGFR-positive SCLC cell lines had the in vitro invasive phenotype, and it is therefore suggested that the EGFR might play an important role for the invasion potential of SCLC cell lines. Images Figure 1 Figure 3 Figure 4

A panel of 21 small cell lung cancer cell (SCLC) lines were examined for the presence of Transforming growth factor beta receptors (TGF beta-r) and the expression of TGF beta mRNAs. By the radioreceptor assay we found high affinity receptors to be expressed in six cell lines. scatchard analysis of the binding data demonstrated that the cells bound between 4.5 and 27.5 fmol mg-1 protein with a KD ranging from 16 to 40 pM. TGF beta 1 binding to the receptors was confirmed by cross-linking TGF beta 1 to the TGF beta-r. Three classes of TGF beta-r were demonstrated, type I and type II receptors with M(r) = 65,000 and 90,000 and the betaglycan (type III) with M(r) = 280,000. Northern blotting showed expression of TGF beta 1 mRNA in ten, TGF beta 2 mRNA in two and TGF beta 3 mRNA in seven cell lines. Our results provide, for the first time, evidence that a large proportion of a broad panel of SCLC cell lines express TGF beta-receptors and also produce TGF beta mRNAs.

Summary The Mr 24, 000 and Mr 52, 000 estrogen-regulated cytosol proteins, and the breast cancer-associated antigen DF3 have been studied in an immunocytochemical assay. Primary tumor specimens from 119 patients with advanced breast cancer who received endocrine therapy have been studied. Monoclonal antibodies were used for the detection of the proteins in formalin-fixed paraffin-embedded blocks. No correlation between Mr 52, 000-positive specimens and the presence of estrogen receptor (ER) could be established (p = 0. 87, chi-square test) whereas a statistically significant association between Mr 24, 000 (p = 0. 0002), DF3 antigen (p = 0. 044) and ER was demonstrated. No intercorrelation was found between Mr 24, 000 and Mr 52, 000 or DF3 (p = 0. 63, 0. 98 and 0. 12 respectively). Clinical response was evaluated for immunocytochemical findings, Mr 24, 000 (p = 0. 37), Mr 52, 000 (p = 0. 61) and DF3 (p = 0. 68) showed no association whereas ER was statistically correlated (p = 0. 00005). Neither overall survival nor disease-free survival correlated to Mr 24, 000 (p = 0. 18 and 0. 75 respectively, logrank test), Mr 52, 000 (p = 0. 095 and 0. 38), or DF3 (p = 0. 22 and 0. 13) staining, whereas ER-positive tumors did (p = 0. 00005). Discrimination between ER-positive re-sponders and ER-positive non-responders was not possible using either Mr 52, 000, Mr 24, 000 or DF3 staining. Based on our findings we conclude that immunocytochemical staining for Mr 52, 000, Mr 24, 000 or DF3 cannot be used as a marker to predict response to endocrine therapy in patients with advanced or recurrent breast cancer.

Hepatic estrogen receptor concentrations were measured in liver biopsy specimens from 102 patients (58 women and 44 men) with liver disease and correlated to several clinical, biochemical, and histologic background variables by means of multiple regression analysis. Half of the tissue was processed for histologic evaluation with a semiquantitative registration of histologic characteristics, whereas the other part was used for receptor analysis by enzyme immunoassay. Fifteen patients with no or minimal histologic changes served as controls. The analysis shows that the reduced estrogen receptor concentrations observed in patients with chronic liver diseases reflect the degree of liver dysfunction and not the specific type of liver disease. Serum albumin, log serum bilirubin, log serum alkaline phosphatases, and the degree of parenchymal fibrosis were significantly related to hepatic estrogen receptor level in the final regression model, but only part of the variation can be explained by these variables, and other factors must be of importance.

We have investigated the expression of cyclins, cyclin dependent kinases (CDK), and CDK inhibitors (CKI) at the mRNA level in a panel of small-cell lung cancer (SCLC) cell lines in vitro and in vivo as xenografts in nude mice. The results showed that the cell lines expressed varying amounts of most cyclin and CDK's but only a few of the cell lines expressed cyclin D1 and/or D2 and some lacked expression of CDK6. Most cell lines expressed mRNA for the CKI's but two cell lines lacked expression of P15INK4B and p16INK4A. The mRNA expression differed for a few of the cell lines regarding cyclin D2 and CDK6 when in vitro and in vivo data were compared. Two of the cell lines that express the retinoblastoma (Rb) protein had no sign of a deregulated Rb pathway but further studies at the protein level are necessary to demonstrate whether these two cell lines should have a normal Rb pathway or whether they will join the majority of cell lines with deregulated Rb pathway.

Transforming growth factor β (TGF-β) exerts a growth inhibitory effect on many cell types through binding to two types of receptors, the type I and II receptors. Resistance to TGF-β due to lack of type II receptor (RII) has been described in some cancer types including small cell lung cancer (SCLC). The purpose of this study was to examine the cause of absent RII expression in SCLC cell lines. Northern blot analysis showed that RII RNA expression was very weak in 16 of 21 cell lines. To investigate if the absence of RII transcript was due to mutations, we screened the poly-A tract for mutations, but no mutations were detected. Additional screening for mutations of the RII gene revealed a GG to TT base substitution in one cell line, which did not express RII. This mutation generates a stop codon resulting in predicted synthesis of a truncated RII of 219 amino acids. The nature of the mutation, which has not previously been observed in RII, has been linked to exposure to benzo[a]-pyrene, a component of cigarette smoke. Since RII has been mapped to chromosome 3p22 and nearby loci are often hypermethylated in SCLC, it was examined whether the lack of RII expression was due to hypermethylation. Southern blot analysis of the RII promoter did not show altered methylation patterns. The restriction endonuclease pattern of the RII gene was altered in two SCLC cell lines when digested with Sma 1. However, treatment with 5-aza-2′-deoxycytidine did not induce expression of RII mRNA. Our results indicate that in SCLC lack of RII mRNA is not commonly due to mutations and inactivation of RII transcription was not due to hypermethylation of the RII promoter or gene. Thus, these data show that in most cases of the SCLC cell lines, the RII gene and promoter is intact in spite of absent RII expression. However, the nature of the mutation found could suggest that it was caused by cigarette smoking. © 1999 Cancer Research Campaign

Our panel of SCLC cell lines have previously been examined for their radiobiological characteristics and sensitivity to treatment with TGF beta 1. In this study we examined the possible correlations between radiobiological parameters and the expression of the TGF beta type II receptor (TGF beta-rII). We have, in other studies, shown that the presence of TGF beta-rII was mandatory for transmitting the growth inhibitory effect of TGF beta. The results showed a statistically significant difference in Dq, i.e. the shoulder width of the survival curve, between cell lines expressing TGF beta-rII and cell lines which did not express the receptor (P = 0.01). Cell lines expressing TGF beta-rII had a high Dq-value. TGF beta-rII expression did not correlate with any other radiobiological parameters. We suggest that an intact growth inhibitory pathway mediated by the TGF beta-rII may have a significant role for the repair of radiation induced DNA damage in SCLC.

Tumor growth is critically dependent on angiogenesis, which is sprouting of new vessels from pre-existing vasculature. This process is regulated by inducers and inhibitors released from tumor cells, endothelial cells. and macrophages. Brain tumors, especially glioblastoma multiforme, have significant angiogenic activity primarily by the expression of the angiogenic factor VEGF. Anti-angiogenic therapy represents a new promising therapeutic modality in solid tumors. Several agents are currently under evaluation in clinical trials. The present review describes the principal inducers and inhibitors of angiogenesis in tumors and summarizes what is known about their mechanisms of action in relation to CNS tumors. Potential areas for clinical use are also discussed.

44 INVOLVEMENT OF THE EPIDERMAL GROWTH FACTOR RECEPTOR IN LARYNGEAL CANCER PATIENTS TREATED WITH HYPOXIA MODIFICATION AS AN ADDITIVE TO ACCELERATED RADIOTHERAPY. M.M. Nijkamp, P.N. Span, I.J. Hoogsteen, C.H.J. Terhaard, P.A.H. Doornaert, J.A. Langendijk, P.L.A. van den Ende, A. Chin, A.J. van der Kogel, J. Bussink, J.H.A.M. Kaanders Department of Radiation Oncology, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands Department of Radiation Oncology, University Medical Centre Utrecht, The Netherlands Department of Radiation Oncology, VU Medical Centre, Amsterdam, The Netherlands Department of Radiation Oncology, University Medical Centre Groningen, The Netherlands Department of Radiation Oncology, Maastro Clinic, Maastricht, The Netherlands Department of Clinical Oncology, Leiden University Medical Centre, Leiden, The Netherlands

The present paper is a comprehensive review of available data concerning the role of radiotherapy as an intended curative treatment in patients with non-small cell lung cancer (NSCL). The following issues are reviewed (1) optimal dose, (2) optimal fractionation, (3) optimal treatment planning, (4) clinical results in terms of single treatment and combined treatment with either surgery or chemotherapy. In resectable NSCLC high dose radiotherapy to small localized tumours gives a 5-year survival rate of 7-38%. It is concluded that this treatment modality is appropriate for certain selected patients who refuse to have surgery, who have medical contradications for surgery, or who are of old age. It is discussed whether the treatment should be split course, continuous, hypo-og hyperfraction. A total dose of 55 Gy must be given. CT scanning should be mandatory for optimal planning and therapy. The literature does not give a conclusive answer to whether preoperative or postoperative radiotherapy is indicated. The data indicate that patients with Stage III NSCLC will benefit from a combined treatment modality in terms of chemotherapy based on high dose cisplatinum and radiotherapy. The main conclusion of the review is that many areas with randomized controlled trials are needed in order to answer the critical issue of the role of radiotherapy in the treatment of NSCLS.

Nine human small-cell lung cancer cell lines were treated with transforming growth factor beta 1 (TGF-beta 1). Seven of the cell lines expressed receptors for transforming growth factor beta (TGF-beta-r) in different combinations between the three human subtypes I, II and III, and two were receptor negative. Growth suppression was induced by TGF-beta 1 exclusively in the five cell lines expressing the type II receptor. For the first time growth suppression by TGF-beta 1 of a cell line expressing the type II receptor without coexpression of the type I receptor is reported. No effect on growth was observed in two cell lines expressing only type III receptor and in TGF-beta-r negative cell lines. In two cell lines expressing all three receptor types, growth suppression was accompanied by morphological changes. To evaluate the possible involvement of the retinoblastoma protein (pRb) in mediating the growth-suppressive effect of TGF-beta 1, the expression of functional pRb, as characterised by nuclear localisation, was examined by immunocytochemistry. Nuclear association of pRb was only seen in two of the five TGF-beta 1-responsive cell lines. These results indicate that in SCLC pRb is not required for mediation of TGF-beta 1-induced growth suppression. Images Figure 2 Figure 3

2052 Background: Concomitant temozolomide (T) and radiotherapy (RT) is standard care in patients (pts) with newly diagnosed glioblastoma (GBM). Bevacizumab (B) and irinotecan (I) produces impressiv...

Recent research shows that concentrations of cytoplasmic estrogen receptors in a breast cancer not simply its presence or absence is correlated with the likelihood of response to endocrine manipulation. This study examines the feasibility of determining estrogen-binding sites in fine-needle aspirations from patients with breast cancer to determine the likelihood of a response to endocrine manipulation. Fine-needle aspiration biopsies were obtained from 60 consecutive females with malignant and benign breast tumors. The fine-needle aspiration technique is described. The following parameters were compared to evaluate the quality of information obtained from fine-needle and biopsy materials from the same tumors: 1) biopsy histology vs. cytology of aspirates; 2) presence/absence of estrogen receptors; 3) Kd and binding capacities in receptor-positive tumors; and 4) estrogen content in relation to protein content. In cytosols with 1 mg/ml or more protein content no significant difference in the number of receptor (+) tumors was observed in fine-needle aspirations as compared to receptor determination of tumor biopsies. Both the fine-needle and corresponding biopsies had comparable binding capacities (fmole bound estrogen/mg protein) but the fine-needle aspirations had significantly lower Kd values. In this study the fine-needle method appears to be inferior to receptor determinations based on tumor biopsy speciments mainly because of lack of available material and the fact that some aspirations are not cytologically representative of the total tumor.

In 46 cases of invasive ductal mammary carcinoma the relationship between estrogen receptor (ER) content and ploidy was investigated. Seventy-one percent of the diploid tumors were ER+ against 40% ER+ tumors in the nondiploid group. Forty percent of the ER+ cases had nondiploid tumors and in several of these more than one tumor cell population could be demonstrated. The results support the hypothesis that multiclonal or mosaic composition of the tumor may explain why 30-45% of the patients with apparently ER+ tumors do not respond to endocrine therapy.

To study the relationship previously noted between bone mass and estrogen bone mineral content (BMC) was determined in the forearm by photon absorptiometry and the amount of estrogen receptors in breast tumor tissues by dextran/charcola estrogen receptor (ER) assay. In all 24 untreated breast cancer patients were measured. BMC and BMC (gm. of ash/cm sq.) expressed as percentage of normal values did not differ from normals neither in ER positivity nor in negativity compared with age- and sex-matched controls. Both BMC (r=.71 P < .001) and BMC (r= -.82 P < .001) decreased significantly with age whereas no significant relation was seen between bone width (BW) and age (r=.06). The amount of ER increased significantly with age (r-.49 P < .02). In ER-positive patients the BMC percentage was significantly elevated compared with receptor-negative patients (P < .01) and a significant positive correlation was found between BMC percentage and the amount of ER in breast cancer tissue (r-.58 P < .01) whereas no significant correlation was found between BMC percentage and the amount of ER. Similarly a significant relationship was found between BMC percentage and the dissociation constant (Kd.P < .01) whereas no significant relation was seen between the BMC percentage and Kd. No relation was found between BW and amount of ER. The BMC and BMC percentages were unrelated to the clinical state of the patients. In sum patients with detectable levels of estrogen binding/mg of tissue protein had significantly higher BMC than did patients with undetectable estrogen binding.

This review will focus on the following topics. (1) Growth factors related to lung cancer and how these relate to other malignancies. (2) Possible interactions between chemotherapy and growth factor action. (3) Future directions

A study was made on the effect of ovariectomy, 17 beta-oestradiol, and tamoxifen on the oestrogen and progesterone receptor-positive T61 human breast carcinoma grown in nude mice. The effect of the treatment was evaluated by the specific growth delay calculated on the basis of Gompertz growth curves, and by the changes in the cell cycle distribution monitored by flow cytometric DNA analysis. The results demonstrated that both oestradiol and tamoxifen induced a temporary growth delay, whereas ovariectomy of the host had no effect on the growth of the tumour. The oestradiol-induced tumour growth delay was accompanied by a decrease in the G1 fraction, an accumulation of cells in the S-phase, and polyploidy, whereas neither treatment with tamoxifen nor ovariectomy influenced cell cycle distribution. The results indicate that oestradiol and tamoxifen have different mechanisms of action. In addition, they were interpreted as indicating different mechanisms regulating ovarian-dependent tumour growth, on the one hand, and oestrogen and tamoxifen-induced tumour growth inhibition, on the other. The results support the view that the presence of receptors may be of importance but is not a sufficiently clear marker to allow prediction of the endocrine sensitivity of individual breast tumours.

A prospective study of 292 consecutive patients with 303 tumours was performed to appraise the combination of physical examination, mammography and fine-needle puncture (triple test) in the diagnosis of breast tumours. The diagnostic accuracy of the triple test was 100%. Because of its relative smallness, however, the series does not permit the conclusion that a benign result of triple test makes excisional biopsy unnecessary. It is emphasized that a follow-up regimen with participation of radiologists and cytologists is required. Fine-needle puncture with cytologic examination of the aspirate is recommended as a routine procedure in the diagnosis of breast tumours.

Bone metastases are frequently one of the first signs of disseminated disease in cancer patients. In the majority of patients, the primary tumour originates from the breast, prostate or lungs. Even although the prognosis is serious, a proportion of the patients will survive for several years and will thus require active treatment. More than 25% of the patients have no symptoms whereas pain dominates in the remainder. Frequent complications are pathological fractures, hypercalcaemia and spinal cord compression. Normally, the diagnosis can be established from the clinical picture compared with a series of laboratory analyses, x-ray investigations of the skeleton and bone scintigraphy. As treatment is mainly palliative, the purpose is primarily relief of pain, prevention of fractures and ensuring a reasonable functional level. The therapeutic possibilities comprise local treatment in the form of surgery and irradiation and also systemic treatment in the form of chemotherapy, endocrine therapy and possibly diphosphonates. of these, irradiation still plays the most important role. About 80% of the patients respond to treatment and, after 12 months, 50-70% of the surviving patients will still be free from pain. Only few randomized investigations are found in the literature available and the criteria of response are, in general, difficult to interpret. There is, therefore, a great requirement for more clinically controlled investigations which assess the efficacy of the numerous therapeutic possibilities.

Samples of human breast cancer tissue were analyzed for estrogen receptor content (ER) with a hydroxylapatite method (HAP) as well as a dextran-coated charcoal method (DCC). It was found that both methods revealed artificially low ER concentrations in cytosols with low protein concentration. It was also found that the DCC assay underestimated the ER activity in the tumors compared to the HAP assay. No differences in terms of low affinity binding nor Kd-values of high affinity estrogen binding were observed. A significant higher background radioactivity was obtained in the HAP assay, and in some tumors one assay would classify them as ER-negative and another as ER-positive.

A patient who had two primary tumors in the same breast is described. Both tumors were infiltrating duct carcinomas, but they differed as far as histologic malignancy and estrogen-receptor-positivity are concerned. One tumor was estrogen-receptor-positive the other was estrogen-receptor-negative. The case illustrates the multicentric origin of breast cancer as well as its cellular heterogeneity. Problems of treatment are discussed.

The ploidy level was investigated by flow cytometric analysis in 143 cases of invasive intraductal mammary carcinoma. Aneuploidy was found in 70% of the tumours. Comparison of ploidy level with histopathologic features, hormone receptor status and clinical characteristics indicated that aneuploid tumours were mostly poorly differentiated (grade III) and oestrogen receptor negative (p less than 0.05). However, this applied only to postmenopausal patients. No correlation appeared between the progesterone receptor status and the ploidy level, but this parameter tended to predict the frequency of lymph node metastases. The possible prognostic significance of these findings is discussed.

The investigation was carried out to elucidate a possible relationship between the amount of estrogen receptors and bone mineral content in patients with breast tumors. Bone mineral content (BMC) was measured by photon absorptiometry in the distal forearm of 54 women with untreated breast carcinoma and 19 with benign breast tumor. The concentration of unoccupied high affinity estrogen receptors was measured in breast tumor biopsy specimens. Higher values of BMC were found in the total group of estrogen receptor-positive patients with breast carcinoma compared with estrogen receptor-negative patients, but not after dividing the patients into smaller groups according to age. No significant correlation could be seen between the amount of estrogen receptors and bone mineral content. In conclusion the present study could not support a relationship between the amount of estrogen receptors in breast cancer tissue and the amount of bone mineral and bone mass in women with breast tumors.