[Tumor biology of oropharyngeal carcinoma]. / Tumorbiologie des Oropharynxkarzinoms.

BACKGROUND: Etiologically, oropharyngeal squamous cell carcinoma (OPSCC) can be divided into OPSCC caused by noxious agents and human papillomavirus (HPV)-driven carcinoma. These types differ with regard to clinical features and prognosis-differences which are rooted in the underlying molecular biology of the tumor. OBJECTIVE: The aim of this work is to provide an overview of the molecular biological characteristics of the genetics, epigenetics, and immunology of OPSCC. MATERIALS AND METHODS: A literature review was performed on a selection of genetic, epigenetic, and immunological factors characterizing OPSCC. RESULTS: The understanding of genetic aberrations and their consequences for cancerogenesis and tumor biology is increasing. Epigenetic phenomena are complementing functional relationships. However, epigenetic mechanisms of gene regulation are complex and much research is still required in this field. Immunological aspects of cancer molecular biology have moved into the focus in light of recent advances in the field of immunotherapy. CONCLUSION: The tumor biology of OPSCC is primarily defined by its HPV status. Additionally, HPV-independent genetic, epigenetic, and immunological signatures are being defined. From these advances, rationales for new treatment concepts may evolve.

RUNX1 mutations in acute myeloid leukemia are associated with distinct clinico-pathologic and genetic features.

We evaluated the frequency, genetic architecture, clinico-pathologic features and prognostic impact of RUNX1 mutations in 2439 adult patients with newly-diagnosed acute myeloid leukemia (AML). RUNX1 mutations were found in 245 of 2439 (10%) patients; were almost mutually exclusive of AML with recurrent genetic abnormalities; and they co-occurred with a complex pattern of gene mutations, frequently involving mutations in epigenetic modifiers (ASXL1, IDH2, KMT2A, EZH2), components of the spliceosome complex (SRSF2, SF3B1) and STAG2, PHF6, BCOR. RUNX1 mutations were associated with older age (16-59 years: 8.5%; ⩾60 years: 15.1%), male gender, more immature morphology and secondary AML evolving from myelodysplastic syndrome. In univariable analyses, RUNX1 mutations were associated with inferior event-free (EFS, P<0.0001), relapse-free (RFS, P=0.0007) and overall survival (OS, P<0.0001) in all patients, remaining significant when age was considered. In multivariable analysis, RUNX1 mutations predicted for inferior EFS (P=0.01). The effect of co-mutation varied by partner gene, where patients with the secondary genotypes RUNX1mut/ASXL1mut (OS, P=0.004), RUNX1mut/SRSF2mut (OS, P=0.007) and RUNX1mut/PHF6mut (OS, P=0.03) did significantly worse, whereas patients with the genotype RUNX1mut/IDH2mut (OS, P=0.04) had a better outcome. In conclusion, RUNX1-mutated AML is associated with a complex mutation cluster and is correlated with distinct clinico-pathologic features and inferior prognosis.

Molecular radiotherapy: the NUKFIT software for calculating the time-integrated activity coefficient.

PURPOSE: Calculation of the time-integrated activity coefficient (residence time) is a crucial step in dosimetry for molecular radiotherapy. However, available software is deficient in that it is either not tailored for the use in molecular radiotherapy and/or does not include all required estimation methods. The aim of this work was therefore the development and programming of an algorithm which allows for an objective and reproducible determination of the time-integrated activity coefficient and its standard error. METHODS: The algorithm includes the selection of a set of fitting functions from predefined sums of exponentials and the choice of an error model for the used data. To estimate the values of the adjustable parameters an objective function, depending on the data, the parameters of the error model, the fitting function and (if required and available) Bayesian information, is minimized. To increase reproducibility and user-friendliness the starting values are automatically determined using a combination of curve stripping and random search. Visual inspection, the coefficient of determination, the standard error of the fitted parameters, and the correlation matrix are provided to evaluate the quality of the fit. The functions which are most supported by the data are determined using the corrected Akaike information criterion. The time-integrated activity coefficient is estimated by analytically integrating the fitted functions. Its standard error is determined assuming Gaussian error propagation. The software was implemented using MATLAB. RESULTS: To validate the proper implementation of the objective function and the fit functions, the results of NUKFIT and SAAM numerical, a commercially available software tool, were compared. The automatic search for starting values was successfully tested for reproducibility. The quality criteria applied in conjunction with the Akaike information criterion allowed the selection of suitable functions. Function fit parameters and their standard error estimated by using SAAM numerical and NUKFIT showed differences of <1%. The differences for the time-integrated activity coefficients were also <1% (standard error between 0.4% and 3%). In general, the application of the software is user-friendly and the results are mathematically correct and reproducible. An application of NUKFIT is presented for three different clinical examples. CONCLUSIONS: The software tool with its underlying methodology can be employed to objectively and reproducibly estimate the time integrated activity coefficient and its standard error for most time activity data in molecular radiotherapy.

Chitinase enzyme activity in CSF is a powerful biomarker of Alzheimer disease.

OBJECTIVE: DNA damage accumulation in brain is associated with the development of Alzheimer disease (AD), but newly identified protein markers of DNA damage have not been evaluated in the diagnosis of AD and other forms of dementia. METHODS: Here, we analyzed the level of novel biomarkers of DNA damage and telomere dysfunction (chitinase activity, N-acetyl-glucosaminidase activity, stathmin, and EF-1α) in CSF of 94 patients with AD, 41 patients with non-AD dementia, and 40 control patients without dementia. RESULTS: Enzymatic activity of chitinase (chitotriosidase activity) and stathmin protein level were significantly increased in CSF of patients with AD and non-AD dementia compared with that of no dementia control patients. As a single marker, chitinase activity was most powerful for distinguishing patients with AD from no dementia patients with an accuracy of 85.8% using a single threshold. Discrimination was even superior to clinically standard CSF markers that showed an accuracy of 78.4% (ß-amyloid) and 77.6% (tau). Combined analysis of chitinase with other markers increased the accuracy to a maximum of 91%. The biomarkers of DNA damage were also increased in CSF of patients with non-AD dementia compared with no dementia patients, and the new biomarkers improved the diagnosis of non-AD dementia as well as the discrimination of AD from non-AD dementia. CONCLUSIONS: Taken together, the findings in this study provide experimental evidence that DNA damage markers are significantly increased in AD and non-AD dementia. The biomarkers identified outperformed the standard CSF markers for diagnosing AD and non-AD dementia in the cohort investigated.

[Current aspects of the pathology and differentiation of extranodal marginal zone B-cell lymphoma, MALT-Type, and gastrointestinal diffuse large B-cell lymphoma]. / Aktuelle molekular-zytogenetische Aspekte zur Pathologie und Differenzierung von extranodalen Marginalzonen B-Zell-Lymphomen vom MALT-Typ und gastrointestinalen diffusen grosszelligen B-Zell-Lymphomen.

The marginal zone B-cell lymphoma, MALT-type (MZBL, MT) is a low-grade B-cell lymphoma which is predominantly localised in the stomach with a typical morphology and cytogenetic pattern. The coexistence of a diffuse large B-cell lymphoma (DLBCL) with an MZBL, MT in the gastrointestinal tract is defined as a composite lymphoma (ComL) and represents a fascinating model of lymphoma progression. In this review we focus on current aspects regarding the molecular characterisation of MZBL, MT and gastrointestinal DLBCL and their mutual relationships.

[A prospective comparison of video colonoscopy and CT colonography in asymptomatic patients screened for colorectal cancer]. / CT-Kolonographie versus Videokoloskopie zum Nachweis kolorektaler Läsionen in der Vorsorgepopulation.

BACKGROUND AND OBJECTIVE: It was the aim of this study to compare the sensitivity and specificity of low-dose CT colonography (CTC) with that of optical colonoscopy (OC) in asymptomatic patients undergoing these tests in a screening program for colonic cancer. PATIENTS AND METHODS: 58 patients (mean age 62.6 years) were included. They underwent low dose CTC and, immediately afterwards, colonoscopy. The colonoscopists were unaware of the CTC findings. A "second look" was performed if a lesion seen in CTC had been missed in the first colonoscopy. RESULTS: A total of 150 lesions were detected and histologically confirmed. 136 were found to be polypoid lesions, classified as either hyperplastic polyps (n = 66) or polyps with intraepithelial neoplasia (n = 70). In the per-patient analysis only 22.4 % of patients had no polypoid lesion, 27.6 % had at least one hyperplastic and 50.0 % had at least one adenomatous lesion. Sensitivity for adenomas of all size categories was calculated 55.7 % for CTC and 92.9 % for OC. This marked difference (both for the detection of individual lesions and the per-patient analyses) does not reach significance in the two-sided McNemar test. CONCLUSIONS: There was a high prevalence of lesions with intraepithelial neoplasia in this screening group. OC had a higher sensitivity than CTC in the detection of lesions smaller than 10 mm.

Patients with in-stent restenoses: comparison of intracoronary beta-brachytherapy using a rhenium-188 filled balloon catheter with the polymer-based paclitaxel-eluting taxus-express stent.

AIMS: We compared the intracoronary beta-brachytherapy using a liquid rhenium-188 filled balloon with the slow-release, polymer-based, paclitaxel-eluting Taxus-Express stent for treatment of in-stent restenoses. PATIENTS, METHODS: During the same study period, patients with restenoses in bare-metal stents were either treated with Taxus-Express stents (n = 50) or beta-brachytherapy after successful angioplasty (n = 51). For brachytherapy 30 Gy in 0.5 mm tissue depth were administered. The irradiated segment exceeded the traumatized segment 7.5 mm on both sides. Primary endpoint was the minimal lumen diameter (MLD) at the target lesion at six months follow-up. Angiographic follow-up was available in 78% (n = 79/101) and clinical follow-up in all patients. RESULTS: Baseline parameters did not differ statistically. The Taxus-Express stent resulted in a significantly larger MLD and a significantly lower percent diameter stenosis post intervention compared to beta-brachytherapy, which both maintained until angiographic follow-up (primary endpoint 2.44 +/- 0.74 mm versus 1.73 +/- 0.74 mm, p < 0.0001). Therefore, Taxus-Express stents were associated with a lower angiographic restenosis rate compared with beta-brachytherapy, both for the target lesion (6.1% versus 17.4%) and the total segment (9.1% versus 23.9%). Moreover, use of Taxus-stent was associated with a clinical benefit based on a significantly lower MACE rate compared with beta-brachytherapy (p < 0.05). CONCLUSIONS: Paclitaxel-eluting Taxus-Express stents resulted in superior clinical and angiographic outcomes compared to intracoronary beta-brachytherapy with a liquid (188)Re filled balloon for treatment of restenosis within a bare-metal stent.

Further delineation of chromosomal consensus regions in primary mediastinal B-cell lymphomas: an analysis of 37 tumor samples using high-resolution genomic profiling (array-CGH).

Primary mediastinal B-cell lymphoma (PMBL) is an aggressive extranodal B-cell non-Hodgkin's lymphoma with specific clinical, histopathological and genomic features. To characterize further the genotype of PMBL, we analyzed 37 tumor samples and PMBL cell lines Med-B1 and Karpas1106P using array-based comparative genomic hybridization (matrix- or array-CGH) to a 2.8k genomic microarray. Due to a higher genomic resolution, we identified altered chromosomal regions in much higher frequencies compared with standard CGH: for example, +9p24 (68%), +2p15 (51%), +7q22 (32%), +9q34 (32%), +11q23 (18%), +12q (30%) and +18q21 (24%). Moreover, previously unknown small interstitial chromosomal low copy number alterations (for example, -6p21, -11q13.3) and a total of 19 DNA amplifications were identified by array-CGH. For 17 chromosomal localizations (10 gains and 7 losses), which were altered in more than 10% of the analyzed cases, we delineated minimal consensus regions based on genomic base pair positions. These regions and selected immunohistochemistries point to candidate genes that are discussed in the context of NF-kappaB transcription activation, human leukocyte antigen class I/II defects, impaired apoptosis and Janus kinase/signal transducer and activator of transcription (JAK/STAT) activation. Our data confirm the genomic uniqueness of this tumor and provide physically mapped genomic regions of interest for focused candidate gene analysis.

Transcriptional profiling suggests that secondary and primary large B-cell lymphomas of the gastrointestinal (GI) tract are blastic variants of GI marginal zone lymphoma.

The pathogenetic relationship of marginal zone B-cell lymphoma (MALT lymphoma) of the gastrointestinal (GI) tract and eventually co-existing aggressive B-cell lymphoma and primary aggressive B-cell lymphoma remains to be elucidated. The RNA of laser-microdissected cells was isolated and amplified from small and/or large cell compartments of eight MALT lymphomas (small cell lymphoma, SCL), 14 GI diffuse large B-cell lymphomas (large cell lymphoma, LCL), and ten GI B-cell lymphomas with composite small and large cell compartments (ComL) and expression analyses were performed using cDNA arrays. Hierarchical cluster analysis clearly separated SCL and LCL and the small and large cell compartments of ComL. Likewise, cluster analysis with all samples of SCL, LCL, and ComL yielded two main 'small cell' and 'large cell' branches. Furthermore, 60 genes were differentially expressed between SCL and LCL, and 82 genes between the small and large cell components of ComL; 26 genes were discriminators in both settings. Use of the profiles of ComL as training sets for class prediction resulted in 95% accuracy for the classification of SCL and LCL. Collectively, the data strongly suggest that both secondary and primary aggressive B-cell lymphomas of the GI tract are blastic marginal zone lymphomas.

Prediction of the axillary lymph node status in mammary cancer on the basis of clinicopathological data and flow cytometry.

Axillary lymph node status is a major prognostic factor in mammary carcinoma. It is clinically desirable to predict the axillary lymph node status from data from the mammary cancer specimen. In the study, the axillary lymph node status, routine histological parameters and flow-cytometric data were retrospectively obtained from 1139 specimens of invasive mammary cancer. The ten variables: age, tumour type, tumour grade, tumour size, skin infiltration, lymphangiosis carcinomatosa, pT4 category, percentage of tumour cells in G2/M- and S-phases of the cell cycle, and ploidy index were considered as predictor variables, and the single variable lymph node metastasis pN (0 for pN0, or 1 for pN1 or pN2) was used as an output variable. A stepwise logistic regression analysis, with the axillary lymph node as a dependent variable, was used for feature selection. Only lymphangiosis carcinomatosa and tumour size proved to be significant as independent predictor variables; the other variables were non-contributory. Three paradigms with supervised learning rules (multilayer perceptron, learning vector quantisation and support vector machines) were used for the purpose of prediction. If any of these paradigms was used with the information from all ten input variables, 73% of cases could be correctly predicted, with specificity ranging from 82 to 84% and sensitivity ranging from 60 to 63%. If only the two significant input variables were used, lymphangiosis carcinomatosa and tumour diameter, the prediction accuracy was no worse. Nearly identical results were obtained by two different techniques of cross-validation (leave-one-out against ten-fold cross validation). It was concluded that: artificial neural networks can be used for risk stratification on the basis of routine data in individual cases of mammary cancer; and lymphangiosis carcinomatosa and tumour size are independent predictors of axillary lymph node metastasis in mammary cancer.

[Necessity and usefulness of bioinformatic methods for microarray data analysis]. / Wertigkeit und Notwendigkeit bioinformatischer Methoden zur Mikroarray-Datenanalyse.

Data emerging from DNA microarray experiments are usually difficult to interpret. While the level of expression of several thousand genes can be measured in a single experiment, only a few dozen experiments are normally carried out, leading to data sets of very high dimensionality and low cardinality. The computational analysis of gene expression data makes significant usage of machine learning and statistical methods. Nevertheless, caution should be used in the blind adoption of these methods, as this usually leads to an over-interpretation of the expression profiles. The following presentation provides an overview of up-to-date principles of biostatistical analysis. A potential application for the analysis of high-dimensional expression profiles of prostate cancer is given.

Incidental carcinoma of the prostate: clinicopathological, stereological and immunohistochemical findings studied with logistic regression and self-organizing feature maps.

OBJECTIVE: To identify significant predictive factors determining category T1a and T1b in incidental prostatic carcinoma with classical and neural multivariate data analysis methods. MATERIALS AND METHODS: Incidental prostatic carcinomas diagnosed in our department during 1990-99 (66 cases) were re-examined. Besides acquiring routine clinical and pathological data the tumours were assessed by scoring immunohistochemistry for proliferative activity and p53-overexpression. Tumour vascularization (angiogenesis) and epithelial texture variables were investigated by quantitative stereology. The data were evaluated by classical statistical methods (t-test, correlation analysis, logistic regression). Moreover, self-organizing feature maps (SOMs) were applied as an exploratory approach to unsupervised data analysis by artificial neural networks. RESULTS: The proliferative fraction, p53 overexpression of tumour cell nuclei, preoperative prostate-specific antigen value and density of capillary vascularization correlated with the Gleason score in incidental prostatic carcinoma. In a stepwise logistic regression analysis with the tumour categories T1a and T1b as dependent variables, the Gleason score and the volume fraction of epithelial cells were significant independent predictors of the tumour category. The cases could be grouped into clusters of different degrees of malignancy using SOMs. CONCLUSIONS: Texture variables of tumour cells are of central importance for the extent of propagation in the prostate in incidental prostatic adenocarcinomas. Gleason score and quantitative stereological estimates of the volume fraction of tumour cells are significant predictors of T1a and T1b categories of incidental prostatic carcinoma. Unsupervised clustering of T1 prostate carcinoma cases by SOMs correlates well with the dichotomous classification into T1a and T1b according to the UICC.

[Effect of molsidomine on rheological parameters and the incidence of cardiovascular events]. / Einfluss von Molsidomin auf rheologische Parameter und die Inzidenz kardiovaskulärer Ereignisse.

BACKGROUND AND OBJECTIVE: In-vitro studies revealed that nitric oxide (NO) may affect rheological parameters. We studied the effect of highly-dosed NO-donor molsidomine on blood rheology and the impact of rheological parameters on the incidence of severe cardiovascular events. PATIENTS AND METHODS: In this randomized, placebo-controlled and double-blind trial 166 patients (60 +/- 10 years) with stable angina pectoris and coronary intervention received molsidomine 3 x 8 mg t. i. d. (controlled release tablets) or placebo for 6 months. Patients with inflammatory/neoplastic disorders or elevated values of C-reactive protein were excluded from analysis. A rheological profile (plasma viscosity, blood viscosity, aggregation and flexibility of erythrocytes, filtrability of leukocytes, fibrinogen levels) was done initially and after 6 months. Adverse cardiovascular events (death, myocardial infarction, stroke, coronary/peripheral revascularization) were recorded during 12 months. Furthermore, the impact of rheological parameters regarding the occurrence of severe cardiovascular events (death, myocardial infarction, stroke) was evaluated during a follow-up of median 38 months. RESULTS: The data of 137 patients (n = 71 placebo, n = 66 molsidomine) were analysed. The difference of rheological parameters between the two measurements did not vary between the two groups. Analysis of event-free survival with Kaplan-Meier technique revealed no difference between the two groups. Multivariate Cox regression analysis with adjustment for diabetes mellitus, smoking and therapy with statin showed a significant association of fibrinogen and plasma viscosity with the occurrence of severe cardiovascular events. CONCLUSION: Treatment with molsidomine 3 x 8 mg/day for 6 months does not improve blood rheology or reduce cardiovascular events. But elevated levels of fibrinogen and plasma viscosity were associated with the occurrence of severe cardiovascular events.

DNA microarray analysis in malignant lymphomas.

Recently, DNA microarray technology has opened new avenues for the understanding of lymphomas. By hybridization of cDNA to arrays containing >10,000 different DNA fragments, this approach allows the simultaneous evaluation of the mRNA expression of thousands of genes in a single experiment. Using sophisticated bioinformatic tools, the huge amount of raw data can be clustered resulting in (1) tumor subclassification, (2) identification of pathogenetically relevant genes, or (3) biological predictors for the clinical course. This approach already has provided novel insights into different entities of B-cell non-Hodgkin's lymphomas. Genomic DNA chip hybridization (matrix-CGH) is a complementary approach focussing on genomic aberrations. In this review, we discuss the impact of this new technology both with regard to methodological aspects as well as to novel findings influencing our understanding of lymphomas.

Prediction of postoperative prostatic cancer stage on the basis of systematic biopsies using two types of artificial neural networks.

OBJECTIVE: The choice of therapy for prostatic cancer should depend on a rational preoperative estimate of tumor stage. Artificial neural networks were used to predict postoperative staging of prostatic cancer from sextant biopsies and routinely available preoperative data. METHODS: In group I (97 cases), nonorgan confinement (tumor stage > or =pT3a) was predicted on the basis of age and six histopathological variables from sextant biopsies. In group II (77 cases), nonorgan confinement and extraprostatic organ infiltration (tumor classification > or =pT3b) were predicted from age, four histopathological variables, the preoperative PSA level, and the total prostate volume estimated by preoperative ultrasonography. Learning vector quantization (LVQ) networks were applied for this purpose and compared to multilayer perceptrons (MLP) and linear discriminant analysis (LDA). RESULTS: Nonorgan confinement could be predicted correctly in 90% of newly presented cases from sextant biopsy histopathology alone. A similar accuracy of predicting nonorgan confinement (83%) was obtained by combining preoperative biopsy histology with clinical data. Extraprostatic organ infiltration could be predicted correctly in 82%. The best results were obtained by LVQ networks, followed by MLP networks and LDA. CONCLUSION: The postoperative tumor stage of prostatic cancer can be estimated with high accuracy, sensitivity and specificity from preoperative routine parameters using artificial neural networks, especially LVQ networks. The results suggest that this methodology should be evaluated in a larger prospective study.

Cluster analysis of comparative genomic hybridization (CGH) data using self-organizing maps: application to prostate carcinomas.

Comparative genomic hybridization (CGH) is a modern genetic method which enables a genome-wide survey of chromosomal imbalances. For each chromosome region, one obtains the information whether there is a loss or gain of genetic material, or whether there is no change at that region. Usually it is not possible to evaluate all 46 chromosomes of a metaphase, therefore several (up to 20 or more) metaphases are analyzed per individual, and expressed as average. Mostly one does not study one individual alone but groups of 20-30 individuals. Therefore, large amounts of data quickly accumulate which must be put into a logical order. In this paper we present the application of a self-organizing map (Genecluster) as a tool for cluster analysis of data from pT2N0 prostate cancer cases studied by CGH. Self-organizing maps are artificial neural networks with the capability to form clusters on the basis of an unsupervised learning rule, i.e., in our examples it gets the CGH data as only information (no clinical data). We studied a group of 40 recent cases without follow-up, an older group of 20 cases with follow-up, and the data set obtained by pooling both groups. In all groups good clusterings were found in the sense that clinically similar cases were placed into the same clusters on the basis of the genetic information only. The data indicate that losses on chromosome arms 6q, 8p and 13q are all frequent in pT2N0 prostatic cancer, but the loss on 8p has probably the largest prognostic importance.

Cardiac vulnerability assessment from electrical microvariability of high-resolution electrocardiogram.

Patients susceptible to malignant arrhythmias often have an increased beat-to-beat variation of the T-wave of the electrocardiogram. Variability analysis of the T-wave is increasingly used for non-invasive risk assessment. The aim of this study is to evaluate intra-QRS beat-to-beat signal variation and to compare it to ST-T variation. The beat-to-beat, microvolt variation of the QRS and the ST-T segment from 44 patients with coronary heart disease at high risk of suffering from malignant arrhythmias and from 51 healthy volunteers are compared. Variation analysis is carried out on 250 consecutive sinus beats from high-resolution electrocardiograms. The individual beats are filtered using a waveform-independent, cubic spline-filter. A variability index of the QRS and ST-T segments is calculated as the integrated standard deviation of corresponding samples inside the area of interest. Patients at risk of suffering from malignant arrhythmias have a significantly higher variability index of both the QRS (median 44.5 ms against 34.7 ms, p < 0.001) and the ST-T segment (median 20.5 ms against 9.8 ms, p < 0.001) compared to the group of healthy subjects. The discriminative ability of the odds variability indices of the QRS and ST-T segments are not statistically different, the ratios being 7.8 (QRS) and 12.6 (ST-T). We conclude that patients at high risk of suffering from malignant arrhythmias are characterised by an increased beat-to-beat microvolt variation of both the QRS and the ST-T segment. Further studies are necessary to evaluate the prognostic potential of depolarisation variability.