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Background: Current risk stratification models for early invasive (T1) colorectal cancer are not able to discriminate accurately between prognostic favourable and unfavourable tumours, resulting in over-treatment of a large (>80%) proportion of T1 colorectal cancer patients. The tumour-stroma ratio (TSR), which is a measure for the relative amount of desmoplastic tumour stroma, is reported to be a strong independent prognostic factor in advanced-stage colorectal cancer, with a high stromal content being associated with worse prognosis and survival. We aimed to investigate whether the TSR predicts clinical outcome in patients with non-pedunculated T1 colorectal cancer., Methods: Haematoxylin and eosin (H&E)-stained tumour tissue slides from a retrospective multicentre case cohort of patients with nonpedunculated surgically treated T1 colorectal cancer were assessed for TSR by two independent observers who were blinded for clinical outcomes. The primary end point was adverse outcome, which was defined as the presence of lymph node metastasis in the resection specimen or colorectal cancer recurrence during follow-up., Results: All 261 patients in the case cohort had H&E slides available for TSR scoring. Of these, 183 were scored as stroma-low, and 78 were scored as stroma-high. There was moderate inter-observer agreement κ = 0.42). In total, 41 patients had lymph node metastasis, 17 patients had recurrent cancer and five had both. Stroma-high tumours were not associated with an increased risk for an adverse outcome (adjusted hazard ratio = 0.66, 95% confidence interval 0.37-1.18; p = 0.163)., Conclusions: Our study emphasises that existing prognosticators may not be simply extrapolated to T1 colorectal cancers, even though their prognostic value has been widely validated in more advanced-stage tumours., (© 2021 The Authors. United European Gastroenterology Journal published by Wiley Periodicals LLC. on behalf of United European Gastroenterology.)

Objective: To investigate whether chorionic villus sampling (CVS) results in a proportional increase of alpha-fetoprotein (AFP) and fetal red cells in maternal blood., Methods: Blood samples were collected before and after CVS. The AFP concentration was measured and supervised automated microscopy of Kleihauer-Betke slides was applied to quantify fetal red cells., Results: AFP analysis was performed in 53 paired samples and automated microscopic scanning in 59 paired samples. Median AFP concentrations before and after CVS were 12.0 microg/L (range 6.4-36.4) and 18.7 microg/L (range 8.2-668.9), respectively, indicating a significant increase (p < 0.0001). Median numbers of fetal red cells detected before and after CVS were 0 (range 0-36) and 0 (range 0-31), respectively. No significant increase of fetal cells was observed (p = 0.72). The delta (Delta) fetal red cells and the Delta AFP correlated poorly (rho = -0.22, p = 0.11). The amount of villi correlated moderately with the Delta AFP (rho = 0.32, p = 0.02) and did not correlate with the Delta fetal red cells (rho = -0.11, p = 0.43)., Conclusions: Although the AFP concentration after CVS increased, no increase of fetal red cells was detected. These findings suggest that CVS results in a leakage of proteins due to placental tissue damage, rather than increased trafficking of fetal cells., (2006 John Wiley & Sons, Ltd.)

We have developed a two-colour immunocytochemical staining method for the detection of fetal and embryonic haemoglobin in erythroid cells. The method was applied to study these haemoglobin types in fetal red cells. Specimens from fetal blood (10 weeks), cord blood and fetal liver (14 weeks) as well as chorionic villus samples (10-13 weeks) were stained for gamma and epsilon chains using CY3 and FITC labelled antibodies. Morphometric analysis was applied to determine cell size. Samples from organs involved in early embryonic development contained relatively large erythroblasts expressing the epsilon globin chain (megaloblasts); later in gestation the gamma chain was co-expressed by the same cells which ultimately became smaller and contained HbF (alpha 2 gamma 2) only. This phenomenon was confirmed in CVS samples in which all cell types were abundantly present. Since fetal erythroblasts are considered candidate cells for non-invasive prenatal diagnosis using FISH, we studied the phenotype of erythroblasts circulating in the maternal blood. The majority of erythroblasts in maternal blood appeared to be of the relatively small gamma globin-containing cell type. However, careful screening of the same maternal blood samples also revealed erythroblasts expressing epsilon or epsilon and gamma globins simultaneously, although at low frequency. Control specimens from non-pregnant women did not show nucleated red cells expressing either of the haemoglobin types. These observations may contribute to the better recognition of fetal cells in the maternal blood for prenatal diagnosis.

In a pilot study to establish fetal nucleated red blood cell (NRBC) detection in maternal blood, trisomy 13 was diagnosed by FISH analysis at 11 weeks' gestation. The NRBCs were detected after a single-step ficoll density gradient enrichment. In blood samples taken both before and after CVS, 52 and 80 NRBCs, respectively, were found to be positive for fetal haemoglobin. In 47 per cent of these cells, FISH analysis for X and Y chromosomes confirmed the fetal sex. Moreover, 48 per cent of these NRBCs showed three fluorescent signals for a chromosome 13 probe, which confirmed the diagnosis of trisomy 13, previously detected at CVS karyotyping. This is the first report of non-invasive prenatal diagnosis of trisomy 13, i.e., pre-CVS, in the first trimester. The high number of fetal NRBCs detected indicates a connection with aneuploidy, probably due to early impairment of the feto-maternal barrier.

One of the major drawbacks in cytomegalovirus (CMV)-antigenaemia detection for diagnosis of active CMV infection is the low number of CMV-antigen positive cells present in peripheral blood. It is therefore necessary to screen large numbers of peripheral blood granulocytes to find only a few antigen-positive cells. We have optimized this detection by testing several monoclonal antibodies (mAb) to CMV-antigens (mAbs C10/C11, C12, BM222, E13 and SL20). In total 550 blood samples from 40 patients were investigated. More blood samples were found positive with mAb C12 than with the other mAbs. Also the average number of positive cells per slide was highest for mAb C12. Furthermore, duplicate slides were examined automatically using an image analysis system (LEYTAS) and compared to visual detection (cytospin slides). The detection sensitivity of both screening methods was compared for mAb C12. In total 360 slides were analysed, from positive as well as negative blood samples. The sensitivity of the automated screening was 93% and for the visual evaluation of the cytospin slides 73%. In conclusion, mAb C12 was the most suitable of the mAbs tested for detection of antigenaemia, and automatic detection of CMV antigenaemia with image analysis of slides is a sensitive method due to the large numbers of cells that can be screened.