Long-term effects of adjuvant tamoxifen and/or radiotherapy. The South Sweden Breast Cancer Trial.

In a multicenter trial of adjuvant therapy in stage II breast cancer, 719 postmenopausal patients were randomized to one of three treatment regimens: radiotherapy only or in combination with adjuvant tamoxifen for one year, or adjuvant tamoxifen without radiotherapy. At twelve years of follow-up (median 9 years), no statistically significant differences in survival or recurrence-free survival were observed. However, the rate of loco-regional recurrency was lower among patients treated with both radiotherapy and tamoxifen. The rate of bilateral breast cancer was reduced in tamoxifen-treated patients whereas the rate of new primary malignancies other than breast cancer was somewhat higher in tamoxifen-treated patients. Adjuvant therapy in breast cancer may influence not only breast cancer recurrences and mortality but also later disease patterns and cause-specific mortality.     

Whole‐exome sequencing defines the mutational landscape of pheochromocytoma and identifies KMT2D as a recurrently mutated gene

As subsets of pheochromocytomas (PCCs) lack a defined molecular etiology, we sought to characterize the mutational landscape of PCCs to identify novel gene candidates involved in disease development. A discovery cohort of 15 PCCs wild type for mutations in PCC susceptibility genes underwent whole‐exome sequencing, and an additional 83 PCCs served as a verification cohort for targeted sequencing of candidate mutations. A low rate of nonsilent single nucleotide variants (SNVs) was detected (6.1/sample). Somatic HRAS and EPAS1 mutations were observed in one case each, whereas the remaining 13 cases did not exhibit variants in established PCC genes. SNVs aggregated in apoptosis‐related pathways, and mutations in COSMIC genes not previously reported in PCCs included ZAN, MITF, WDTC1, and CAMTA1. Two somatic mutations and one constitutional variant in the well‐established cancer gene lysine (K)‐specific methyltransferase 2D (KMT2D, MLL2) were discovered in one sample each, prompting KMT2D screening using focused exome‐sequencing in the verification cohort. An additional 11 PCCs displayed KMT2D variants, of which two were recurrent. In total, missense KMT2D variants were found in 14 (11 somatic, two constitutional, one undetermined) of 99 PCCs (14%). Five cases displayed somatic mutations in the functional FYR/SET domains of KMT2D, constituting 36% of all KMT2D‐mutated PCCs. KMT2D expression was upregulated in PCCs compared to normal adrenals, and KMT2D overexpression positively affected cell migration in a PCC cell line. We conclude that KMT2D represents a recurrently mutated gene with potential implication for PCC development. © 2015 The Authors. Genes, Chromosomes & Cancer Published by Wiley Periodicals, Inc.     

A phase I/II evaluation of metoclopramide as a radiosensitiser in patients with inoperable squamous cell carcinoma of the lung

The feasibility of administering metoclopramide (MCA) as a radiosensitizer has been evaluated in 23 patients with a pathological or cytological diagnosis of a squamous cell carcinoma of the lung, clinically evaluated as inoperable. All patients received 40–60 Gy radiotherapy fractionated into 1.8 Gy fractions 5 times per week (Monday–Friday). Two MCA treatment regimens were used: (i) MCA at 2 mg/kg administered by intravenous infusion 1–2 h prior to radiotherapy 3 times per week (Monday, Wednesday, Friday); and (ii) MCA at 1 mg/kg administered by intravenous infusion 1–2 h prior to radiotherapy 5 times per week (Monday–Friday). 11 of the 23 patients treated with radiotherapy and MCA had none to mild pneumonitis or fibrosis and another 8 of the 23 had moderate levels. No patient had their therapy interrupted due to radiation-related side-effects. The MCA-related side-effects were as expected, i.e. 78% of the patients experienced sedation/tiredness and 48% expressed restlessness/anxiety symptoms. Both the total dose and serum levels of MCA were significantly associated to the MCA side-effect profile. Tumour response, duration of tumour response and survival were significantly positively correlated to the total and weekly doses of MCA administered to the patients during their radiotherapy treatment. These favourable phase II data have justified the initiation of a phase II/III randomised multicentred trial being carried out in Europe to evaluate MCA as a radiosensitiser.     

Comparative cytogenetic and dna flow cytometric analysis of 150 bone and soft-tissue tumors

Samples from 48 benign and 102 malignant bone and soft-tissue tumors were analyzed cytogenetically and by DNA flow cytometry. Clonal chromosome abnormalities were found in 82 tumors and normal karyotypes in 68; 61 tumors were DNA-non-diploid and 89 were diploid. The cytogenetically abnormal tumors were used for comparison between the 2 types of investigation; 45 of these tumors were DNA-diploid and 37 were DNA-non-diploid. There was, with few exceptions, good correspondence between the quantitative estimates of genomic changes by the 2 methods, indicating that the cells cytogenetically analyzed from short-term cultures are representative of the in vivo cell populations. Discrepancies were primarily found in cases with indexes above 1.5, in which the DNA index was higher than the chromosome index. The chromosome analysis suggested that skewed stemline (G₀/G₁) peaks in the diploid region in DNA histograms indicate the presence of cell populations with small net quantitative genomic changes, although not all such populations were detected by DNA flow cytometric analysis. The view that one of the peaks in bimodal stemline DNA histograms with narrow peaks represents a non-diploid cell population was also corroborated. On average, the cell populations giving rise to double stemlines in DNA histograms showed quantitatively larger genomic changes than those that gave rise to broad or skewed diploid G₀/G₁ peaks. The findings indicate that these histogram profiles are not artifactual but reflect chromosomal changes in the tumor parenchyma.     

Increase in intracellular immunoglobulins in the majority of splenic lymphoid cells after primary immunization

The amount of immunoglobulin (Ig) in individual mouse spleen lymphocytes was determined by quantitative microspectrofluorimetry after “staining” with fluorescein-labeled anti-Ig antibodies. During a primary immune response against sheep erythrocytes, maximally 0.015 % of the cells produced direct hemolytic plaques, but close to 100 % of lymphoid cells displayed an increase in their intracellular Ig on day 3 to 4 after immunization. This increase consisted of IgM, later to be followed in a large fraction (up to 80 %) of the cells by IgGl. A positive correlation was found between rate of protein synthesis and fluorescence intensity in the single cell. Preliminary data suggested that activated thymus-derived cells displayed an increase in an intracellular material, cross-reactive with serum IgM. Attempts to demonstrate that the intracellular increase was due to endocytosis of Ig in various forms failed in several model systems. The reported phenomenon might demonstrate the production of a substance, capable of alerting the majority of all lymphoid cells during a primary immune response, into increased Ig synthesis.     

Humoral immunity targeting site I of antigenic domain 2 of glycoprotein B upon immunization with different cytomegalovirus candidate vaccines

Glycoprotein B (gB) is a major component in several vaccines that are under development for prevention of disease by cytomegalovirus. It contains multiple determinants that are targets for neutralizing antibodies. One of them is site I of antigenic domain 2 (AD-2). The epitope, defined by short peptides, is quite conserved between different isolates. However, it is poorly immunogenic in natural infection. In this study we investigated the extent to which different vaccines, attenuated live Towne vaccine with or without priming with a canarypox virus coding for gB, or a recombinant gB vaccine adjuvanted with MF59, induced antibodies to this epitope. As in natural infection only a fraction of all subjects developed antibody responses against site I of AD-2 following vaccination. We suggest that strategies that enhance immunogenicity of this epitope will improve vaccine efficacy.     

Prognostic potential of flow cytometric S-phase and ploidy prospectively determined in primary breast carcinomas

In a prospective study of a consecutive breast cancer series accumulated in the period 1978–82, the S-phase fraction (SPF) and ploidy status were determined by flow cytometry performed on cell nuclei derived from samples of 580 primary tumors. Sixty percent of the tumors were non-diploid. After correction for debris the median SPF values were 7.3% overall, 12% for non-diploid tumors, and 2.9% for diploid tumors (2.6% when nodal subsets N2 and N3 and cases with metastases at presentation were excluded). The SPF values correlated both to tumor size (p=0.008) and to the number of positive axillary lymph nodes (p=0.03).At clinical follow-up in 1986, 467 unilateral breast cancer patients who had undergone radical treatment for cure could be evaluated with respect to the prognostic value of both the SPF value and ploidy status. The median duration of follow-up was then 59 months (range 2–90), and the median time-to-recurrence 24 months (range 2–69, n=137).At follow-up in 1991, 201/467 of the patients had died, the median duration of follow-up being 50 months (range 2–126) for the deceased, and 119 (range 6–148) for the survivors. In multivariate analysis (Cox's proportional hazards models), the strongest independent predictors of distant recurrence-free survival (DRFS) were the number of positive axillary lymph nodes (p<0.0001), the debris-corrected SPF value alone (p=0.003,versus p=0.05 for uncorrected value), and ploidy status combined with the corrected SPF value (p=0.0002). When age was taken into account, both the corrected SPF value and the ploidy-SPF combination were predictors of crude survival (p=0.006 and p=0.002, respectively).In univariate life-table analysis, the 5-year DRFS rate was 93% in node-negative (N0) cases with an SPF<7.3%, as compared to 80% in those with an SPF7.3% (p=0.005). Among node-positive cases, the prognostic value of the SPF was confined to those with 1–3 positive nodes, the 5-year DRFS rate being 68% in cases with an SPF<7.3%, as compared to 40% in cases with an SPF7.3% (p=0.01).Ploidy status and SPF were combined to form four groups: diploid & SPF<2.6% (DL), diploid & SPF2.6% (DH), non-diploid & SPF<12% (NDL), and non-diploid & SPF12% (NDH). Among node-negative patients, the DRFS rate fell from 95% in the DL group to 87% in the NDL group, with the DH group at an intermediate level, as compared with 74% (p=0.03) for the NDH group which accounted for the bulk of the early distant recurrences. Among patients with 1–3 positive lymph nodes, the 5-year DRFS rate was 68% in both the groups with low SPF values (DL and NDL), as compared with 45% in the DH group (p=0.03), and 37% in the NDH group (p=0.006).In this study, the flow cytometry SPF value, alone or in combination with ploidy status, yielded the most profound additional prognostic information, enabling both node-negative patients with a high probability of cure and patients at risk of early relapse to be identified. Among node-positive patients, the prognostic value of the SPF value was confined to those with 1–3 positive axillary lymph nodes (the predominant node-positive subgroup), enabling a high and a low DRFS rate subgroup to be distinguished – a useful distinction where selection for adjuvant drug treatment is concerned. As the predictive strength of the SPF value was enhanced when correction was made for debris, we would recommend that the effect of such factors as debris be minimized as far as possible when flow cytometry-derived SPF values are to be used for prognostic purposes.     

Efficient reduction of loco-regional recurrences but no effect on mortality twenty years after postmastectomy radiation in premenopausal women with stage II breast cancer – A randomized trial from the South Sweden Breast Cancer Group

PurposeTo study long term loco-regional and distant recurrence rate and survival after post-mastectomy radiotherapy in combination with oral cyclophosphamide in premenopausal women with stage II breast cancer.Study designA three-armed randomized multicenter phase III trial comparing 1) Radiotherapy (RT) 2) RT+ oral cyclophosphamide for one year (RT + C) and 3) Oral cyclophosphamide only (C).Radiotherapy was administered, in 20 fractions, to 48 Gy to the axilla and parasternal lymph nodes, 45 Gy to infra- and supraclavicular fossae and 38 Gy to the chest wall. Cyclophosphamide was prescribed as 12 courses of 130 mg/m² od for14 days every 4 weeks.Patients and methods367 patients from 15 surgical departments in Southern Sweden, representing 80% of all eligible patients, were included in the trial between 1978–1983. Median age was 47 years, median tumour size was 25 mm, and 33% of the patients were lymph node negative. Median follow-up time was 24 years.ResultsRT reduced the risk at twenty years for loco-regional recurrence in C-treated patients at twenty years with 75% (13.9% vs. 3.5%). The risk reduction was highly significant in both N0 and N+ patients. No reduction in systemic disease or mortality was observed.ConclusionPost-mastectomy radiotherapy reduced loco-regional recurrences in this premenopausal population, but no effect was seen on mortality with 20 years follow-up.     

Evaluation of cerebrospinal fluid (CSF) C-reactive protein in the diagnosis of suspected meningitis

Cerebrospinal fluid C-reactive protein (CSF-CRP) was studied in 183 consecutive infants and children with suspected meningitis, using a nephelometric technique. Cerebrospinal fluid C-reactive protein was above an empirically chosen level of 1 mg/1 in seven of 19 children with culture-proven bacterial meningitis, in only one of 15 children with viral meningitis, and three of 139 children with no meningitis. All 10 children with partially treated meningitis had CSF-CRP levels below 1 mg/1. There was good correlation between CSF-CRP and total protein levels in children with bacterial meningitis (R value 0.4999 P less than 0.05). The test was not sensitive enough for early differentiation between bacterial and viral meningitis. The test also did not add extra information regarding aetiology in partially treated meningitis.