Commentary on a combined approach to the problem of developing biomarkers for the prediction of spontaneous preterm labor that leads to preterm birth.

INTRODUCTION: Globally, preterm birth has replaced congenital malformation as the major cause of perinatal mortality and morbidity. The reduced rate of congenital malformation was not achieved through a single biophysical or biochemical marker at a specific gestational age, but rather through a combination of clinical, biophysical and biochemical markers at different gestational ages. Since the aetiology of spontaneous preterm birth is also multifactorial, it is unlikely that a single biomarker test, at a specific gestational age will emerge as the definitive predictive test. METHODS: The Biomarkers Group of PREBIC, comprising clinicians, basic scientists and other experts in the field, with a particular interest in preterm birth have produced this commentary with short, medium and long-term aims: i) to alert clinicians to the advances that are being made in the prediction of spontaneous preterm birth; ii) to encourage clinicians and scientists to continue their efforts in this field, and not to be disheartened or nihilistic because of a perceived lack of progress and iii) to enable development of novel interventions that can reduce the mortality and morbidity associated with preterm birth. RESULTS: Using language that we hope is clear to practising clinicians, we have identified 11 Sections in which there exists the potential, feasibility and capability of technologies for candidate biomarkers in the prediction of spontaneous preterm birth and how current limitations to this research might be circumvented. DISCUSSION: The combination of biophysical, biochemical, immunological, microbiological, fetal cell, exosomal, or cell free RNA at different gestational ages, integrated as part of a multivariable predictor model may be necessary to advance our attempts to predict sPTL and PTB. This will require systems biological data using "omics" data and artificial intelligence/machine learning to manage the data appropriately. The ultimate goal is to reduce the mortality and morbidity associated with preterm birth.

Tetranectin positive expression in tumour tissue leads to longer survival in Danish women with ovarian cancer. Results from the 'Malova' ovarian cancer study.

The primary objective of this study was to analyse Tetranectin (TN) expression in tumour tissues and TN serum concentration in 758 women with epithelial ovarian tumours. The second was to evaluate, whether TN tissue expression levels correlate with clinico-pathological parameters and prognosis of the disease. Using tissue arrays we analysed the expression levels in tissues from 166 women with borderline ovarian tumours (BOTs) and 592 women with ovarian cancer (OC). A panel of three antibodies was used for immunohistochemistry: a polyclonal and two monoclonal antibodies. Serum TN was measured using the polyclonal antibody A-371. Univariate survival analyses stratified for chemotherapy showed that positive tissue TN as demonstrated by the polyclonal antibody indicated a significantly longer overall survival (OS) (p = 0.0001) as well as cancer specific survival (CSS) (p < 0.0001). High serum TN was likewise found to imply longer OS (p < 0.0001) and CSS (p < 0.0001), whereas tissue staining with the two monoclonal antibodies failed to demonstrate any significant correlation with either survival type. Univariate Kaplan-Meier survival analysis performed on all OC cases showed a significantly longer OS (p = 0.0009) and CSS (p = 0.0006) for women with TN positive tumour tissue and in women with high serum TN levels (p < 0.0001 for both). However, in the multivariate Cox regression analysis, only serum TN was found to be an independent prognostic factor for OS (p = 0.01) and not for CSS (p = 0.08). In conclusion, our results predict that a positive TN expression of both tumour tissue and serum points to a more favourable outcome for OC patients.

Prdm5 suppresses Apc(Min)-driven intestinal adenomas and regulates monoacylglycerol lipase expression.

PRDM proteins are tissue-specific transcription factors often deregulated in diseases, particularly in cancer where different members have been found to act as oncogenes or tumor suppressors. PRDM5 is a poorly characterized member of the PRDM family for which several studies have reported a high frequency of promoter hypermethylation in cancer types of gastrointestinal origin. We report here the characterization of Prdm5 knockout mice in the context of intestinal carcinogenesis. We demonstrate that loss of Prdm5 increases the number of adenomas throughout the murine small intestine on an Apc(Min) background. By using the genome-wide ChIP-seq (chromatin immunoprecipitation (ChIP) followed by DNA sequencing) and transcriptome analyses we identify loci encoding proteins involved in metabolic processes as prominent PRDM5 targets and characterize monoacylglycerol lipase (Mgll) as a direct PRDM5 target in human colon cancer cells and in Prdm5 mutant mouse intestines. Moreover, we report the downregulation of PRDM5 protein expression in human colon neoplastic lesions. In summary, our data provide the first causal link between Prdm5 loss and intestinal carcinogenesis, and uncover an extensive and novel PRDM5 target repertoire likely facilitating the tumor-suppressive functions of PRDM5.

Risk of biochemical recurrence and positive surgical margins in patients with pT2 prostate cancer undergoing radical prostatectomy.

BACKGROUND AND OBJECTIVE: To investigate risk factors associated with positive surgical margins (PSM) and biochemical recurrence (BR) in organ confined tumors (pT2) after radical prostatectomy (RP) for localized prostate cancer (PCa). METHODS: Between 1995 and 2011, 1,649 patients underwent RP at our institution. The study includes the 1,133 consecutive patients with pT2 tumors at final histopathology. Logistic regression analysis was used for risk of PSM. Risk of BR, defined as the first PSA ≥ 0.2 ng/ml, was analyzed with Kaplan-Meier and Cox regression analysis. RESULTS: Median follow-up was 3.6 years (range: 0.5-15.5 years). In logistic regression, NS surgery was independently associated with an increased risk of pT2 PSM (OR = 1.68, 95% CI: 1.3-2.0, P = 0.01) relative to non-NS surgery. NS surgery was not independently associated with BR but the interaction of PSM and NS surgery trended (P = 0.08) to increase the risk of BR compared to PSM and non-NS surgery. CONCLUSION: Several factors influence the risk of pT2 PSMs in radical prostatectomy. In our cohort pT2 PSM is associated with NS surgery and trend to increase risk of BR compared to non-NS surgery. The optimal selection of candidates for NS surgery is still not clear.

In vitro and in vivo evaluation of the indoloquinone EO-9 (NSC 382 459) against human small cell carcinoma of the lung.

As the indoloquinone EO-9 has previously shown activity in several tumor model systems it was evaluated against four human small cell lung cancer cell lines by the clonogenic assay. In two cell lines (Nyh and Tol), exponential dose-response curves were achieved with both 1 h and continuous exposure, whereas no cell kill was obtained in the other two cell lines (69 and 592) when tested with 1 h incubation up to 0.25 microgram/ml. When the cells were exposed to drug in vitro, flow cytometric DNA analysis showed perturbations in the cell cycle distribution of the most sensitive cell line (Tol) at a lower EO-9 concentration than in the less sensitive cel line (592). This in vitro predicted difference in EO-9 sensitivity between two of the cell lines (592 and Tol) was confirmed when the cell lines were heterotransplanted to nude mice.

Interactions between three subpopulations of Ehrlich ascites tumor and a P388 murine leukemia in mixed solid tumors in immune competent mice.

Cellular interactions between three subpopulations of Ehrlich ascites tumor and between these and the P388 murine leukemia were studied during growth of solid tumors obtained by mixtures of the cells in immune competent N/D mice. An immunogenic Ehrlich cell line (E1.15) induced an immunologically based growth inhibition of the two other Ehrlich cell lines (E1.80 and E1.95) which themselves were non-immunogenic. E1.15 was, however, unable to induce an immunological response against the P388 cell line. It is therefore suggested that when in close contact, immunologically induced cellular responses imposed by an immunogenic cell line on other cell lines require genetic and thereby close immunogenic resemblance between the cell lines. Another type of interaction was found between the E1.95 cell line and the P388 line which showed nearly identical growth characteristics as determined by tumor weight day 14, tumor growth curves, cell cycle times (per cent labelled mitoses) and cell cycle distributions (flow cytometric DNA analysis). After 2 weeks of growth of mixed P388/E1.95 tumors, flow cytometric DNA analysis on fine-needle tumor aspirates showed nearly total dominance of P388. This type of interaction required close cellular contact of viable cells, and no cellular immune response was elicited by the host animals. A third finding was that a faster growing Ehrlich cell line E1.95 dominated the tumors when inoculated in mixture with a slower growing subpopulation E1.80. This could be explained on the basis of the cell kinetic differences between these two cell lines.

Inter-experiment variation and dependence on culture conditions in assaying the chemosensitivity of human small cell lung cancer cell lines.

Sensitivity of five human small cell lung cancer cell lines to doxorubicin was assessed by a double layer agar technique using two different bottom-layers. Neither of the bottom-layers provided proportionality between numbers of cells plated and numbers of colonies, but they were correlated by a logarithmic function. Even after correction for lack of proportionality the two assay systems provided significantly different dose-response curves. The stability of the chemosensitivity was tested after 25-30 weeks continuous in vitro culture or prolonged storage in liquid nitrogen. One cell line underwent significant changes after continuous in vitro culture whereas the cell lines tested after prolonged storage in liquid nitrogen showed only minor changes. It is concluded that instead of considering the concentration necessary to achieve a certain degree of cell kill (e.g. ID50) in one experiment on one cell line, dose-response curves obtained on several cell lines in different assay systems should be used in the evaluation of new drugs.

Prognostic factors in primary mucinous breast carcinoma.

Two hundred seven mucinous breast carcinomas were morphologically classified into two different groups: pure mucinous carcinomas consisting only of areas with small epithelial islands of solid tumor floating in abundant extracellular mucin, and mixed carcinomas where the tumor contains large areas of mucin, as well as areas with infiltrating carcinoma devoid of extracellular mucin. Of the 207 tumors, 95 were of the pure type and 112 of the mixed type. The pure and mixed carcinomas differed significantly with respect to a number of prognostic factors, the most important of which was lymph node status. Patients with pure mucinous carcinomas had significantly fewer lymph node metastases at the time of primary operation (P = 0.0001) and a significantly longer recurrence-free survival (P = 0.03) than patients with mixed carcinomas. Patients with mixed carcinomas closely resembled patients with infiltrating ductal carcinomas not otherwise specified (NOS) with respect to both lymph node status and recurrence-free survival. In multivariate analysis, the single most important factor for predicting recurrence-free survival among the 207 patients was lymph node status. However, of 94 pure carcinomas, only 6 had lymph node metastases at the time of mastectomy. A detailed morphologic analysis demonstrated that two of these six cases were incorrectly diagnosed as being pure mucinous carcinomas--they were actually of the mixed type. In another two of these cases, the metastases originated from a co-existent infiltrating ductal carcinoma. Thus, metastases to the regional lymph nodes were observed in only two cases of pure mucinous carcinomas. It is concluded that the biologic behavior of the pure mucinous carcinomas differs from that of both the mixed carcinomas and infiltrating ductal carcinomas NOS. Thus, a new definition is suggested, in which a mucinous carcinoma is classified as such only if it is a pure carcinoma, and mixed mucinous carcinomas are classified according to the component without extracellular mucin production.