Effect of surgery on tumour-directed leucocyte responses.

Leucocytes from 22 out of 26 patients with mammary carcinoma were significantly cytotoxic in vitro for cells cultured from mammary tumours though only two out of 17 of these preparations were cytotoxic for cells cultured from tumours arising at other sites. In the immediate postoperative period reactivity of patients'' leucocytes with mammary tumour cells was undetectable but returned within one week of surgery. Leucocyte cytotoxicity may therefore offer a model in which the mechanism of postoperative depression of immunological competence may be investigated.     

Heterogeneity of DNA ploidy in gastric cancer.

Heterogeneity of DNA content was analyzed in 389 samples from 65 resected gastric cancers. Analysis of the samples revealed that there were 14 homogeneously diploid tumours. Six tumours were uniformly DNA aneuploid, each tissue block containing the same DNA index. The other 45 tumours (69%) varied in DNA content heterogeneity. In 39 of 45 tumours, there was a mixture of diploid and aneuploid samples, and 25 of the 39 tumours had a single aneuploid stemline. In 14 out of 39 tumours, there was also a mixture of diploid and aneuploid samples having two or more DNA aneuploid stemlines. In the remaining six tumours, different DNA aneuploid stemlines were contained in different samples without evidence of diploidy. When four or fewer samples were analyzed, only 50% of the tumours were diagnosed as having DNA content heterogeneity. On the other hand, 78% of the tumours showed DNA heterogeneity when 5 or more samples were analyzed. If the tumours had not been widely sampled, about a quarter of the tumours would have been mislabeled as diploid. The patients with tumours showing homogeneous diploidy survived longer than those with tumours showing a mixture of diploid and aneuploid stemlines. The survival rate was lowest for the patients with tumours having a mixture of diploid and multiple aneuploid stemlines, compared with those showing homogeneous diploid or a mixture of diploid and single aneuploid stemlines. The data from the current study clearly demonstrate the importance of adequate sampling in assessing the ploidy status of gastric cancers to identify groups of patients running different clinical course and prognosis.     

Use of tumour marker immunoreactivity to identify primary site of metastatic cancer.

OBJECTIVES--To determine whether variations in the expression of tumour related antigens can predict the origin of tumours. DESIGN--Immunohistological study of tumour marker expression in primary adenocarcinomas and respective metastatic deposits. Antibodies to the following tumour markers were used: polymorphic epithelial mucin (NCRC-11 and SM3), carcinoembryonic antigen, carcinoembryonic antigen with non-specific antigen co-specificity, CA125, CA19.9, prostate specific antigens, and thyroglobulin. SETTING--Histopathology department of teaching hospital. SUBJECTS--100 pathology sections of metastatic adenocarcinoma and their related primary tumours. MAIN OUTCOME MEASURES--Concordance of reactivity between primary and metastatic tumours. Reactivity profiles of tumour sites. RESULTS--The correct primary site of origin was predicted in 70% (33/47) of tumours in men and 54% (27/43) tumours in women with antibodies SM3, 288, CA19.9, CA125, and PSA (men only). Specificities ranged from 68% for breast tumour to 98% for prostate tumour. CONCLUSION--Use of tumour markers in patients presenting with metastatic adenocarcinoma of unknown origin can help localise the probable primary sites and reduce the need for extensive and expensive imaging techniques.     

Localisation of malignant germ-cell tumours by external scanning after injection of radiolabelled anti-alpha-fetoprotein.

Sheep IgG antibody to alpha-fetoprotein was labelled with 131I and used to identify human germ-cell tumours by emission scanning. Eleven patients were studied after resection of their primary tumours. Ten had malignant teratoma and one an endodermal sinus tumour. All eight patients with raised serum alpha-fetoprotein concentrations had metastases apparent in the antibody scans. Of the remaining three patients with normal serum alpha-fetoprotein concentrations, two had positive scans. Three of the patients with positive results were scanned twice; the second scans were negative after treatment, when the alpha-fetoprotein concentrations had returned to normal. These result suggest that antibody scans are useful in the clinical management of patients with germ-cell tumours.     

Epidermal growth factor receptors in breast cancer: association with early relapse and death, poor response to hormones and interactions with neu

Epidermal growth factor receptors (EGFRs) were measured in 221 primary breast cancers by ligand binding with 125I-labelled EGF, and high-affinity sites were quantitated. There was a highly significant inverse relationship between oestrogen receptor (ER) and EGFR (15 EGFR-positive [EGFR+]ER+ and 92 EGFR-negative [EGFR-]ER+: 54 EGFR- ER- and 60 EGFR+ ER-). The relapse-free survival and overall survival were significantly shorter for EGFR+ vs EGFR- tumours (P less than 0.001) by about 2 yr in the case of relapse-free survival. When ER- tumours were substratified by EGFR status, the EGFR- ER- tumours had a prognosis almost as good as the ER+ tumours. In 31 of 184 cases, high expression of neu, correlating with amplification, was found. Expression of neu conferred similar poor prognosis to EGFR expression in all prognostic subgroups. Coexpression of neu and EGFR had an additive adverse effect. Epidermal growth factor receptors (EGFR) and oestrogen receptors (ER) were analysed in 221 patients with primary operable breast cancer by means of radioligand assays. After median follow-up of 24 months (range 3-60 months), there had been recurrences in 99 patients, of whom 72 (median age 56 yr, range 32-77 yr) received tamoxifen alone as first-line treatment for recurrence. 14 patients (19%) showed a response to this therapy and 58 (81%) did not. Of 32 ER+ tumours, 12 (37.5%) showed an objective response to tamoxifen compared with only 2 of 40 (5%) ER- tumours (P less than 0.005). Of 35 EGFR+ tumours, 3 (8.5%) achieved an objective response compared with 11 of 36 (30%) EGFR tumours (P less than 0.05). Only 1 of 28 EGFR+, ER- tumours achieved an objective response. Including patients whose disease remained stable for more than 6 months with the responders, however, EGFR status was a better predictor of response to tamoxifen; 15 of 37 EGFR- patients and 5 of 35 EGFR+ patients responded (P less than 0.01).     

Treatment of Advanced Tumours of Head and Neck with Fast Neutrons

Fast neutrons interact with matter in a different way from x and gamma rays. They have been used at Hammersmith Hospital for the past four years in the treatment of advanced tumours in several sites of the body, and the results of this treatment in the first 100 cases of tumours of the head and neck are described here. Altogether 62 patients who had been referred for fast neutron therapy because it was thought that no other treatment would be effective experienced complete regression of the tumours, and only two recurred. Tumours of the buccal cavity and salivary glands responded particularly well and the relief of pain and ulceration was striking. Side effects were not serious and did not differ from those seen with supervoltage radiation, apart from the reaction of the skin. Follow-up was short, however, owing to deaths from metastases, and out of 76 patients treated more than one year previously only 30 survived. Cases which have not metastasized must therefore be treated so that the effects on tumours and adjacent normal tissues can be observed for several years after treatment. The results obtained so far indicate that it is now justifiable to use neutrons in such cases.     

Trial of thyroid autotransplantation in patients with Graves' disease whose remnant thyroid has unintentionally been made too small at subtotal thyroidectomy

Autotransplantation of thyroid tissue was carried out in 5 patients with Graves' disease in order to prevent postoperative hypothyroidism, because the amount of remnant thyroid tissue was estimated to be too small, i.e. from 3 to 5 g. Approximately 0.5 to 2 g of thyroid tissue was cut into small pieces and transplanted into the sternocleidomastoid muscles or the strap muscles. Although the postoperative serum TSH levels were normal or slightly elevated, the serum concentrations of triiodothyronine were within the normal range in these 5 patients at a follow-up study carried out 2 to 7 years after surgery. Thyroid scanning with I-123 or 99mTc-pertechnetate (Tc-99m) revealed radioisotope uptake at the sites of transplantation in 4 of the 5 patients. These findings verify that the implanted thyroid tissues were alive and functioning and that autotransplantation may be a way of preventing postoperative hypothyroidism in patients whose remnant thyroid tissue has unintentionally become too small.     

Cytological diagnosis of soft tissue tumours

Cytological diagnosis of soft tissue tumours The aims of this study were to determine the patterns of soft tissue tumours and also to try to assess the utility of fine needle aspiration cytology (FNAC) in diagnosing soft tissue tumours. Of 15 361 patients who visited the cytology diagnostic service of the Pathology Department, Medical Faculty, Addis Ababa University, 623 (4.1%) cases with a diagnosis of soft tissue tumours were retrieved from the department's records for the years 1991-96. Fifty-three soft tissue tumours (25 benign and 28 malignant tumours) with combined FNAC and surgical biopsy results were traced for cyto-histological correlations. Twenty-two out of 25 benign soft tissue tumours were correctly diagnosed, with three false cytologic diagnoses where one mesenchmal neoplasm, one haemangioma, and one haemorragic lesion were identified; and out of 28 malignant soft tissue, 23 were correctly diagnosed however, the five false cytological diagnoses were one soft tissue sarcoma, one dermatofibrosarcoma, one malignant mesenchymal neoplasm, one spindle cell neoplasm and one menechymal neoplasm. Thus, in this study a sensitivity and specificity of 88.5% and 81.5% respectively for the diagnosis of soft tissue tumours were reported. In conclusion, FNAC of soft tissue tumours is a fast, effective and reliable diagnostic tool that may help in categorizing soft tissue tumours into benign and malignant groups for clinical management.     

Importance of FNAC in the detection of tumours within multinodular goitre of the thyroid

The primary challenge in the management of a multinodular thyroid gland is to rule out malignancy. The present study was undertaken to assess the value of preoperative ultrasound-guided fine needle aspiration cytology (FNAC) in diagnosing tumours of the thyroid gland. Of the 80 patients operated for multinodular lesions, malignant tumours were found in 29 and benign tumours in 36 patients (81%) and non-tumorous lesions in 15 (19%) patients. Compared with the histopathological postoperative diagnosis, the overall sensitivity of FNAC was 85% and specificity 88%. Current morphological diagnosis of the nodules in multinodular goitre requires thorough preoperative examination, including ultrasound-guided FNAC in order to establish the appropriate management.     

Problems in assessing multiple cutaneous melanoma. A review on the accuracy of a population based cancer registry

Databases with information on malignant tumors are of great value for epidemiologic studies. From the Regional South Swedish Tumour Registry, which is of documented high quality, 24 patients out of 8008 with reported melanoma diagnosis 1973–2003 were reported as having multiple (≥3) primary, invasive cutaneous malignant melanomas (CMM). Of the 76 tumours identified in these patients, 7 (9%) were found not to be invasive melanomas. Additional cases could be put into question since the lesions could be interpreted as epidermotropic metastases, a diagnosis which can be difficult to establish reliably by microscopic examination. Among the 24 patients we could also identify 8 (10%) additional lesions representing invasive CMM, not included in the Tumour Registry database. Thorough information concerning an earlier melanoma diagnosis and its site of presentation is needed from the clinician and the pathologist for optimal assessment of the histology and the prognostication of the patient, as well as proper reporting to a tumour registry. Identifying multiple primary malignant melanomas is also of special importance for counselling patients belonging to families with hereditary disease. In this study it is shown that diagnosing and reporting multiple malignant melanomas can be problematic due to insufficient communication and to the rare and deceptive capability of cutaneous metastases to imitate primary tumours.     

Radiological and Pathological Features Associated with IDH1-R132H Mutation Status and Early Mortality in Newly Diagnosed Anaplastic Astrocytic Tumours

Background

Glioblastoma can occur either de novo or by the transformation of a low grade tumour; the majority of which harbor a mutation in isocitrate dehydrogenase (IDH1). Anaplastic tumours are high-grade gliomas that may represent the final step in the evolution of a secondary glioblastoma or the initial presentation of an early primary glioblastoma. We sought to determine whether pathological and/or radiological variables exist that can reliably distinguish IDH1-R132H-positive from IDH1-R132H-negative tumours and to identify variables associated with early mortality.

Methods

Patients diagnosed with anaplastic astrocytic tumours were included. Magnetic resonance imaging was performed and immunohistochemistry was used to identify tumours with the IDH1-R132H mutation. Survival was assessed 12 months after diagnosis. Variables associated with IDH1-R132H status were identified by univariate and ROC analysis.

Results

37 gliomas were studied; 18 were positive for the IDH1-R132H mutation. No tumours demonstrated a combined loss of chromosomes 1p/19q. Patients with IDH1-R132H-positive tumours were less likely to die within 12 months of diagnosis (17% vs. 47%; p=0.046), more likely to have tumours located in the frontal lobe (55% vs. 16%; p=0.015), and have a higher minimum apparent diffusion coefficient (1.115 x 10^-3 mm²/sec vs. 0.838 x 10^-3 mm²/sec; p=0.016), however, these variables demonstrated only moderate strength for predicting the IDH1-R132H mutation status (AUC=0.735 and 0.711, respectively). The Ki-67 index was significantly lower in IDH1-R132H-positive tumours (0.13 vs. 0.21; p=0.034). An increased risk of death was associated with contrast-enhancement ≥ 5 cm³ in patients with IDH1-R132H-positive tumours while edema ≥ 1 cm beyond the tumour margin and < 5 mitoses/mm² were associated with an increased risk of death in patients with IDH1-R132H-negative tumours.

Conclusions

IDH1-R132H-positive and -negative anaplastic tumours demonstrate unique features. Factors associated with early mortality are also dependent on IDH1-R132H status and can be used to identify patients at high risk for death.     

A combination of neutral loss and targeted product ion scanning with two enzymatic digestions facilitates the comprehensive mapping of phosphorylation sites

We propose here a new strategy for the exhaustive mapping of phosphorylation sites in the Xenopus laevis Cdc25 phosphatase, which regulates cell cycle progression in eukaryotic cells. Two different MS analyses in a linear IT were used to identify the phosphorylated residues. First, a data-dependent neutral loss (DDNL) analysis triggered the fragmentation of peptides that show enhanced neutral loss of phosphoric acid. Second, a targeted product ion scanning (TPIS) mass analysis was carried out in which MS2 events are triggered for specific m/z values. Full coverage of the protein sequence was obtained by combining the two analyses with two enzymatic digestions, trypsin and chymotrypsin, yielding a comprehensive map of the phosphorylation sites. Previous reports have shown Cdc25C to be phosphorylated by Cdc2-cyclin B at four residues (Thr48, Thr67, Thr138 and Ser205). By using this combination of scan modes, we have identified four additional phosphorylation sites (Thr86, Ser99, Thr112 and Ser163) in a recombinant Cdc25C protein containing 198 residues of the NH2-terminal noncatalytic domain. The sensitivity of this combined approach makes it extremely useful for the comprehensive characterization of phosphorylation sites, virtually permitting complete coverage of the protein sequence with peptides within the mass detection range of the linear IT.     

Identification of a human ribosomal protein mRNA with increased expression in colorectal tumours

A human ribosomal protein cDNA was selected from a normal colon cDNA library on the basis of overexpression in familial adenomatous polyposis. Nucleotide sequence analysis was used to identify this cDNA as corresponding to the human equivalent of the rat ribosomal protein L31 (HL31). We have quantified the expression of HL31 mRNA in colorectal tumours and found overexpression in 23 out of 23 cases. Our results indicate that HL31 is associated with a malfunction of normal growth regulatory mechanisms in these tumours, and suggest a role for HL31 in proliferation and neoplasia.     

Radiolabeled polyvinyl alcohol particles: A potential agent to monitor embolization procedures

Polyvinyl alcohol sponge (PVA) is a widely used angiographic embolic agent. The radiolabeling of PVA can accurately identify particle localization and may decrease the possibility of patient morbidity from embolization to distal sites. We incorporated ^99mTc sulfur colloid (SC) into PVA by heating. Animal experiments demonstrated the in vivo stability of the ^99mTc SC-PVA complex and the efficacy of external imaging. ^99mTc SC-PVA biodistribution data and external NaI(Tl) scintillation probe counts were performed, to assess anatomic localization. Embolization with this complex was performed in a patient.     

First results of a randomized clinical trial of fast neutrons compared with X or gamma rays in treatment of advanced tumours of the head and neck. Report to the Medical Research Council.

Results of the first randomized clinical trial to compare the effects of fast neutrons and those of x or gamma rays (photons) in treating patients with advanced tumours of the head and neck are reported. In 37 out of 52 patients treated with neutrons and 16 out of 50 treated with photons the local tumour completely regressed; the tumour later recurred in nine of the 16 photon patients but in none of the 37 neutron patients. The advantages to the neutron-treated patients were seen in tumours of well and poorly differentiated histology and in each site. Complications after treatment did not differ significantly between the groups. Despite these substantial differences in local control of the tumour there were no significant differences in mortality between the series. A detailed study of the effective doses and the response of tumours and normal tissue in each series indicated that the improved results from neutron therapy were due to differences in the biological quality of the beam and not to the rather higher average effective dose in the neutron series. To assess the long-term effects of neutron treatment patients in earlier stages of disease and with smaller tumours should be included in the next phase of the trial.     

Scintillation Scanning of Lungs in Preoperative Assessment of Carcinoma of Bronchus

Lung scans with the use of macroaggregated human serum albumin labelled with technetium-99m were carried out in 52 patients before thoracotomy.Forty-three patients had carcinoma of the bronchus. Tumours less than 2 cm. in diameter on the chest radiograph were not detected. Larger tumours showed defects in perfusion, ranging in size from the mass seen on the chest radiograph to almost absent perfusion of the entire lung. The extent of the defect in perfusion was closely related to involvement of the pulmonary vessels at the hilum by distortion, compression, or invasion by the tumour. Bronchial obstruction played a less important part in producing the defects.The larger the defect in perfusion the greater was the involvement of the hilar and mediastinal structures and the more extensive was the surgery required. When perfusion of the affected lung was less than one-third of the total the tumour was found to be unresectable.     

A molecular cytogenetic approach to studying platinum resistance

The technique of comparative genomic hybridisation (CGH) has until recently been used to screen for common genomic abnormalities in fresh tumour material; it has identified previously unrecognised regions of amplification associated with poor prognosis subtypes of breast cancer and lymphoma. Our group has applied this technique to resistant cell lines and their sensitive counterparts in order to define chromosomal abnormalities associated with acquired drug resistance. We have demonstrated the applicability of this technique to the study of drug resistance using cell lines with known mechanisms of resistance. The ability to detect novel genomic alterations in cell lines with novel mechanisms of resistance was also demonstrated. We subsequently examined the CGH profiles of seven different cell lines made resistant to three platinum analogues and showed the most consistent abnormalities to involve over-representation of regions 4q and 6q. More recently, we have applied the CGH technique to a series of testicular germ cell tumours (TGCTs) collected as formalin-fixed paraffin-embedded biopsy specimens from patients, both pre- and post-therapy using a platinum-based regimen (POMB/ACE). Previous reports have shown over-representation of X, 7q, 8q and 12p and loss of 13q to occur in 25% of primary TGCTs. Over-representation of 12p was confirmed in the majority of these biopsy samples; deletion of 13q was noted in the initial biopsies of several patients. We also demonstrated alterations of 4p, 4q, 5q and 6q in this series of patients. Newly acquired deletions of 2q and 18q and amplifications of 8q were frequently observed in post-chemotherapy samples from resistant tumours. The CGH studies on these patients with TGCT will not only enable us to correlate our observations on clinical material with those from long-term cell lines, but should also identify sites of key genes involved in clinical platinum resistance.     

Calculation of tumour and normal tissue biological effective dose in 90Y liver radioembolization with different dosimetric methods

PurposeRadioembolization with ⁹⁰Y microspheres is an effective treatment for unresectable liver tumours. Two types of microspheres are available: resin (SIR-Spheres®) and glass (Theraspheres®). The aim of this study is to compare biological effective dose (BED) values obtained with three different dosimetric methods.Methods29 HCC patients were included in this study: 15 were treated with resin(mean injected activity 1.5 GBq, range 0.8–2.7 GBq) and 14 with glass microspheres (2.6 GBq, range 1.3–4.1 GBq). Average doses to tumours and normal liver tissues were calculated with AAPM, multi-compartmental MIRD and Voxel-based methods and consequently the BED values were obtained. Planar images were used for the AAPM method: ^99mTc-MAA SPECT-CT attenuation and scatter corrected images (resin) and ⁹⁹m Tc-MAA SPECT attenuation corrected (glass) were employed for the other two methods.ResultsRegardless of type of microspheres, both for tumours and normal liver tissues, no significant statistical differences were found between MIRD and Voxel for both doses and BED values. Conversely AAPM gave discordant results with respect to the other two methods (Mann-Whitney p-values ⩽ 0.01). For resin spheres the calculated tumour-to-normal tissue ratios on planar images were on average 14 times greater than those obtained on SPECT-CT images, while they were 4 times greater on glass. A linear correlation was observed between MIRD and Voxel BEDs.ConclusionsThe AAPM method appears to be less precise for absorbed dose and BED estimation, while MIRD and voxel based dosimetry are more confident each other.     

(99m)Tc-labeling of ciprofloxacin and nitrofuryl thiosemicarbazone using fac-[(99m)Tc(CO)3(H2O)3] core: evaluation of their efficacy as infection imaging agents

The aim of this study was to radiolabel ciprofloxacin (Cip) and nitrofuryl thiosemicarbazone (NFT) with the fac-[(99m)Tc(CO)(3)(H(2)O)(3)](+) core and to evaluate the ability of the radiopharmaceuticals as tracers in detecting sites of infection. Cip and NFT were radiolabeled with the fac-[(99m)Tc(CO)(3)(H(2)O)(3)](+) core and characterized by RHPLC. The stabilities of the preparations were evaluated in saline and rat serum. In vitro binding studies of the radiopharmaceuticals with S. aureus were performed. Biodistribution studies were conducted at different time points after injecting (i.v.) the radiopharmaceuticals in rats (intramuscularly infected with S. aureus) as well as in rats with sterile inflammation. To assess the infection targeting capacity of (99m)Tc-tricarbonyl ciprofloxacin and nitrofuryl thiosemicarbazone, (99m)Tc(v)O-Cip and (99m)Tc(v)O-NFT were used as control. Scintigraphic imaging studies of tricarbonyl compounds and (99m)Tc(v)O-Cip were performed at 4 h after injection. The radiochemical purities of (99m)Tc(CO)(3)-Cip and (99m)Tc(CO)(3)-NFT were between 97-98% as determined by thin layer chromatography (TLRC) and RHPLC; no further purification is necessary before injection. The radiopharmaceuticals exhibited substantial stability when incubated in isotonic saline and serum up to 24 h. Biodistribution studies showed maximum uptake in the infected rat thigh muscle at 4 h post injection and washing out at slower rate from the infected site than the oxo technetium chelate. The mean ratios of uptake in infected/non-infected thighs were 3.87:1, 3.41:1 and 3.17:1 for (99m)Tc(CO)(3)-Cip, (99m)Tc(CO)(3)-NFT and (99m)Tc(v)O-Cip respectively. During scintigraphic studies, infection sites appeared quite distinctly with (99m)Tc(CO)(3)-Cip and (99m)Tc(CO)(3)-NFT, comparable to the behaviour with (99m)Tc(v)O-Cip. These results encouraged us for further development of infection imaging radiopharmaceuticals based on the (99m)Tc-tricarbonyl core.     

Loss of heterozygosity in hypotriploid cell cultures from testicular tumours

Summary We have established cell lines with a hypotriploid chromosome number from four testicular tumours. Each line had at least one Y chromosome and most of the informative centromere and enzyme markers were heterozygous implying that the tumours originated from germ cells before the first meiotic division. The small metacentric marker chromosome (i12p), specific for testicular tumours, was present in all tumour cell lines and up to three copies were found in some lines. Rearrangements of chromosome 1 and 11 were each found in three out of four tumours. The rearrangements of chromosome 1 all resulted in duplication of 1q and deletion of short-arm material from the same chromosome giving loss of heterozygosity for enzyme markers on 1p. Loss of satellite material from chromosome 13 and the centromere region of chromosome 9 were found in single cases. This study shows that even where the chromosome number of tumour cells is near triploid, regions of the genome can be deleted. The chromosomes most frequently involved in rearrangements, 1, 11, and 12 all contain sites of ras oncogenes and it is suggested that loss of normal alleles could result in homozygosity for mutant oncogenes which may play a part in tumour progression.