DNA flow cytometry in stage IB and II cervical carcinoma

In a retrospective study the prognostic significance of nuclear DNA content was investigated, as measured by flow cytometry, of the tumor specimens from 212 women with nonpretreated FIGO stage IB and II cervical cancer. One-hundred and thirty cases (62%) were found to be diploid, whereas 82 (38%) were aneuploid. Univariate analysis of the follow-up data showed an increased relative risk (RR) for recurrence free survival (RFS) for stage II tumors (RR = 1.87, 95% CI: 1.13–3.10, P = 0.015) and for age (RR = 1.52, 95% CI: 0.66–3.52 and RR = 2.35, 95% CI: 1.19–4.65, P = 0.032). Ploidy showed a relative risk of 1.33 (95% CI: 0.83–2.13, NS). In addition, univariate analysis of overall survival (OS) revealed similar results. For the subgroup of patients with primary surgery ( n = 151), positive pelvic nodes (RR = 5.38, 95% CI: 2.70–10.71, P = 0.0001) and parametrial extension (RR = 2.53, 95% CI: 1.24–5.17, P = 0.011) were significant factors for OS after univariate analysis, the estimated effects on RFS were slightly smaller. Multivariate analysis of RFS for the whole study population showed age, histologic grade and stage with a slightly increased risk, but no effect was significant. Ploidy with an RR of 0.97 (95% CI: 0.58–1.62) seems to have no influence on prognosis. For the subgroup with primary surgery, ploidy again failed statistical significance with an RR of 1.20 (95% CI: 0.58–2.49). Our results suggest that abnormalities of the nuclear DNA content in this homogeneous group of patients are associated with clinical and morphological prognosticators, however, ploidy is not an independent prognostic factor for RFS, or for the whole study population or for the subgroup with primary surgery.     

European seroepidemiology network 2: Standardisation of assays for seroepidemiology of varicella zoster virus.

BACKGROUND: The aim of the European Sero-Epidemiology Network (ESEN2) is to harmonise the serological surveillance of vaccine-preventable diseases in Europe. OBJECTIVE: To allow comparison of antibody prevalence in different countries by standardising results into common units. STUDY DESIGN: For varicella zoster virus (VZV), a reference laboratory established a panel of 148 samples, characterised by indirect enzyme-immunoassay (ELISA), indirect immunofluorescence, and complement fixation test. Fifty-seven samples were also studied by the fluorescence antibody to membrane antigen test. The geometric mean of the antibody activity (GMAA) obtained from four ELISA determinations was used to characterise each sample of the panel as positive (GMAA: >100 mIU/ml), equivocal (GMAA: 50-100 mIU/ml) or negative (GMAA: <50 mIU/ml) for antibody to VZV (anti-VZV). Thirteen laboratories, using five different ELISA tests, tested the panel. RESULTS: Agreement with the reference laboratory was above 85% in all cases, and the R(2) values obtained from regression analysis of the quantitative results were always higher than 0.87. Finally, the regression equations could be used to convert national values into a common unitage. CONCLUSION: This study confirmed that results for anti-VZV obtained by different ELISA methods can be converted into common units, enabling the comparison of the seroprevalence profiles obtained in the participant countries.     

Therapy of small breast cancer - four-year results of a prospective non-randomized study

Background: In the early 1980s breast preservation was a rarely applied therapeutic modality in the primary treatment of breast cancer in the Federal Republic of Germany. Reports coming from retrospective studies as well as preliminary results from randomized trials made it desirable to introduce breast preservation in the form of a controlled clinical trial. Study design: In stage pT1 N0 M0 breast cancer, mastectomy as the standard treatment was to be compared with radiotherapy of the remaining breast tissue. The study design originally planned as a randomized trial had to be changed into a prospective observation study due to the low randomization rate. Univariate analysis of prognostic variables was the first step to a valid treatment comparison. Those factors determined as being significant were included together with the treatment effects in a multivariate analysis. A high therapeutic standard was guaranteed by quality control. Results: 1036 out of 1119 recruited patients are evaluable. After a median follow-up of 48 months the following preliminary results can be reported. With the exception of death without recurrence from breast cancer, the 143 events are evenly distributed among the two treatment groups. Locoregional recurrence of the whole patient population was 5%. Out of all prognostic factors examined only tumor size and grading are significant in regard to recurrent disease. Recurrence-free survival decreased in cases with uncertain tumor margins, whereas the width of the margin had no influence on recurrent disease. There was no significant difference in quality of life between the two treatment groups. Conclusions: The four-year results of this study are in accordance with those of other breast preservation trials: There is no significant difference between the two treatment groups in the occurrence of locoregional failure. Incomplete tumorectomy has a negative influence on recurrence. Quality of life seems more dependent on the acceptance of the therapy by the patient than on the therapeutic modality itself. Breast preservation can also be performed appropriately in smaller institutions if the therapeutic standard is guaranteed by quality control.This study was sponsored by the Ministry of Research and Technology (BMFT) of the Federal Republic of Germany.     

Assessment and comparison of prognostic classification schemes for survival data.

Prognostic classification schemes have often been used in medical applications, but rarely subjected to a rigorous examination of their adequacy. For survival data, the statistical methodology to assess such schemes consists mainly of a range of ad hoc approaches, and there is an alarming lack of commonly accepted standards in this field. We review these methods and develop measures of inaccuracy which may be calculated in a validation study in order to assess the usefulness of estimated patient-specific survival probabilities associated with a prognostic classification scheme. These measures are meaningful even when the estimated probabilities are misspecified, and asymptotically they are not affected by random censorship. In addition, they can be used to derive R(2)-type measures of explained residual variation. A breast cancer study will serve for illustration throughout the paper.     

High-resolution sonography of the abnormal cranial suture

Objective. The purpose of this investigation is to elucidate the sonographic features of abnormal major cranial sutures. Materials and methods. Eight excised synostosed suture specimens were evaluated. The high-resolution sonographic appearance was correlated with the histological section, plain radiographs, CT and MRI. Diastatic and molded sutures were also evaluated with sonography and compared with the normal cranial suture appearance. Results. Synostosed sutures demonstrated one or more of the following features: (a) loss of echo-poor fibrous gap between bony plates (five sagittal and coronal synostoses); (b) irregular thickened inner sutural margin (three lambdoid synostoses); (c) loss of bevelled edge (one lambdoid synostosis); (d) asymmetric anterior fontanelle (one coronal synostosis). Cranial molding results in an overlap of echogenic bony plates. Sutural width (the distance between bony plates) is increased in cases of elevated intracranial pressure. Conclusion. Sonography is an inexpensive, radiation-free modality which can confirm synostosis versus molding versus an underlying intracranial lesion as a cause of plagiocephaly. The high-resolution sonographic images also provide a relatively easy means to assess sutural width and may provide information in regard to increased intracranial pressure. Received: 24 March 1997 Accepted: 25 September 1997     

Interaction of treatment with a continuous variable: simulation study of power for several methods of analysis

In a large simulation study reported in a companion paper, we investigated the significance levels of 21 methods for investigating interactions between binary treatment and a continuous covariate in a randomised controlled trial. Several of the methods were shown to have inflated type 1 errors. In the present paper, we report the second part of the simulation study in which we investigated the power of the interaction procedures for two sample sizes and with two distributions of the covariate (well and badly behaved). We studied several methods involving categorisation and others in which the covariate was kept continuous, including fractional polynomials and splines. We believe that the results provide sufficient evidence to recommend the multivariable fractional polynomial interaction procedure as a suitable approach to investigate interactions of treatment with a continuous variable. If subject‐matter knowledge gives good arguments for a non‐monotone treatment effect function, we propose to use a second‐degree fractional polynomial approach, but otherwise a first‐degree fractional polynomial (FP1) function with added flexibility (FLEX3) is the method of choice. The FP1 class includes the linear function, and the selected functions are simple, understandable, and transferable. Furthermore, software is available. We caution that investigation of interactions in one dataset can only be interpreted in a hypothesis‐generating sense and needs validation in new data. Copyright © 2014 John Wiley & Sons, Ltd.     

Detection of Epstein-Barr virus nucleic acid sequences in non-Hodgkin's lymphoma

Tumor tissue specimens of 25 patients with various entities of Non-Hodgkin's lymphomas were examined for the presence of Epstein-Barr virus (EBV) nucleic acids and EBV nuclear antigens (EBNA) by in situ hybridization and anticomplement immunofluorescence. In tumor cells of five patients EBV nucleic acid and EBNA were demonstrated. The histopathologic examinations revealed in these cases one Burkitt's lymphoma, two centroblastic lymphomas and each one lymphoplasmocytic and lymphocytic lymphoma.     

Seroprevalence of hepatitis E virus (HEV) in humans living in high pig density areas of Germany

An increase in acute autochthonous hepatitis E virus (HEV) infections has been recorded in Germany. These are suspected to be zoonotically transmitted from wild boar, deer and domestic pig. The latter may represent a major reservoir for HEV. In this study, 537 sera from humans living in Westphalia and Lower Saxony, representing areas of high pig density in Germany, were tested for the presence of HEV-specific antibodies. Among them were 302 individuals with occupational, direct contact to pigs and 235 individuals without direct contact to pigs. Two commercial tests and one in-house assay were applied for the detection of HEV-specific immunoglobulin G (IgG) antibodies. Sera were also tested in an assay that detects all classes of HEV-specific antibodies. Depending on the test used, the seroprevalence ranged from 4.1 to 27.9 %. Exposition to pigs was found to be associated with a significantly higher seroprevalence in subjects with contact to pigs (13.2–32.8 %) compared with that in non-exposed humans (7.7–21.7 %). In particular, individuals younger than 40 years with occupational exposure exhibited a markedly higher HEV seroprevalence compared with non-exposed individuals of that age group. In general, HEV seroprevalence increased with age resulting in a similar prevalence level in the age group of ≥50 years for exposed and non-exposed individuals. Analysis of all sera by a commercial anti-HEV IgM ELISA revealed 35 positive and 25 borderline samples. However, only one positive serum could be confirmed by an IgM line assay. Selected samples from IgM and/or IgG as well as total HEV antibody-positive individuals were also tested for the presence of HEV RNA. In one of the 78 samples, the only IgM ELISA positive and IgM line assay confirmed sample, RNA of HEV genotype 3 was detected. This sequence has high similarity to HEV sequences obtained from wild boars and domestic pigs from Germany and The Netherlands. This study demonstrates that in addition to the consumption of raw or undercooked meat, direct contact to pigs has to be considered as an additional risk factor for HEV infection.     

Sample size considerations for the evaluation of prognostic factors in survival analysis

When the role of a new prognostic factor is investigated, careful planning of an appropriate study is required. This includes an assessment of the power of the study in terms of sample sizes. An adequate analysis of the independent prognostic effect of a new factor has to be adjusted for the existing standard factors. With survival time as endpoint this will usually be done with the Cox proportional hazards model. Sample size and power formulae in survival analysis have been developed by Schoenfeld for randomized treatment comparisons. In the analysis of prognostic factors the covariates included are expected to be correlated with the factor of primary interest. In this situation, the existing sample size and power formulae are not valid and may not be applied. In this paper, Schoenfeld's formula is first extended to the situation where a correlated factor is included in the analysis. The validity of the resulting approximate asymptotic formula is investigated for its asymptotic behaviour by numerical integration and for its finite behaviour by simulation. Second, an approximate formula for sample size and power is provided to detect an interaction between the interesting and a second correlated factor. This extends the formula for independent effects. Finally, the approach is illustrated by an example on the prognostic impact of DNA ploidy and other factors in advanced ovarian cancer.