Malte Barten

Tumour budding as prognostic factor in stage I/II colorectal carcinoma

Aims : The term tumour ‘budding’ has been coined for the detachment of tumour cells from the neoplastic glands of adenocarcinomas and is presumed to be an early step in the metastatic process. A limited number of studies have shown budding to be an adverse prognostic factor.

Postoperative Chemotherapy May Not Be Necessary for Patients With ypN0-Category After Neoadjuvant Chemoradiotherapy of Rectal Cancer

PurposeAfter neoadjuvant radiochemotherapy and surgery, there is no general agreement about whether postoperative chemotherapy is necessary. With the help of clinical and pathohistologic data, prognostic factors were determined as a basis for the decision to spare a patient additional chemotherapy or to urgently recommend it.ResultsNinety-five patients treated with neoadjuvant 5-fluorouracil-based radiochemotherapy (November 4, 1997 and June 15, 2004) without distant metastases and an R0 (microscopically complete) resection were evaluated. Adjuvant chemotherapy (5-fluorouracil or 5-fluorouracil/folinic acid) was given to 65 of 95 patients (68.4 percent). The disease-free survival rate after 36 months was chosen as the target parameter (median follow-up, 36 months).MethodsThe five-year survival rate for all patients was 80.3 ± 5.6 percent; the five-year disease-free survival was 78.1 ± 5.1 percent; the five-year local control rate was 94.2 ± 5.1 percent. In the univariate and multivariate analysis of the disease-free survival, the pathohistologic lymph node status after radiochemotherapy (ypN) was the only significant prognostic parameter. Disease-free survival (36 months) for patients without lymph node metastases (ypN0) was excellent, independent of whether they had received postoperative chemotherapy (n = 43; 87.5 ± 6.0 percent) or not (n = 29; 87.7 ± 6.7 percent). Patients with ypN2 status have, despite chemotherapy, a poor disease-free survival at 30 ± 17.6 percent after 36 months.ConclusionsThese retrospective data suggest that, for some patients, postoperative chemotherapy can be spared. For patients with ypN2 status, an intensification of the postoperative chemotherapy should be considered. Further evaluation in prospective studies is urgently recommended.

Human papillomavirus 6/11, 16 and 18 in oral carcinomas and benign oral lesions

Abstract. We have examined 118 oral squamous cell carcinomas, 72 oral leukoplakias, 12 cases of cheilitis and 65 of oral lichen planus for the presence of human papillomavirus (HPV) 6/11, 16 and 18 DNA by PCR/Southern blot hybridization. HPV DNA were found in 51/118 carcinomas (43.2%), in 16/72 (22.2%) leukoplakias, 3/12 (25.0%) cheilitic lesions and 10/65 (15.4%) lichen planus cases. These differences were even stronger when analyzing separately for the high-risk types HPV 16 and 18 as compared to low-risk types 6/11. HPV 16 and 18 DNA were present in 41/118 (34.7%) oral carcinomas, 12/72 (16.7%) leukoplakias, 2/12 (16.7%) cheilitic lesions and 6/65 (9.2%) lichen planus. In contrast to this, oral carcinomas displayed the lowest HPV 6/11 detection rate (4.2%), compared with 11.1% for leukoplakias, 8.3% for cheilitic lesions and 7.7% in lichen planus. These results indicate a successive increase of the detection rate of HPV 16 and 18 from low level in non or questionably preneoplastic lesions (lichen planus) to preneoplastic lesions (leukoplakia and cheilitis) and to oral carcinoma. In conclusion, our results suggest an association of oral carcinogenesis and infection with the high-risk HPV types 16 and 18.

P53 mutational spectra are different between squamous-cell carcinomas of the lip and the oral cavity

We studied the p53 mutational spectra of 34 lip and 60 intra‐oral squamous‐cell carcinomas and examined possible etiological and prognostic correlations for these tumor sites. For the p53 analysis of exons 5–8, we used PCR/TGGE screening followed by DNA sequencing. Mutations were found in 18/34 (53%) lip and 22/60 (38%) intra‐oral carcinomas. The p53 mutational spectrum of the intra‐oral carcinomas comprised transitions and transversions in nearly equal frequency (11 to 10). In comparison, transitions were 3.5 times more frequent than transversions (14 to 4) in carcinomas of the lip. The predominant types of base change found in intra‐oral tumors were G:C‐to‐T:A transversions and G:C‐to‐A:T transitions (32% each), while in lip tumors G:C‐to‐A:T transitions (70%) were the most frequent. The rate of lip tumors with mutations was higher in non‐smokers (8/13) than in smokers (9/20). In contrast, p53 mutations in intra‐oral tumors clustered in smokers (18/47 vs. 2/10). G:C‐to‐T:A transversions, regarded as tobacco smoke–associated in lung cancer, were found in 2 moderate and 4 heavy smokers with intra‐oral cancer. This base substitution was found in none of our lip cancers. In lip tumors, a high rate of mutations occurred at dipyridine sites (13/18); among these were 8 C‐to‐T transitions and 1 CC‐to‐TT tandem base transition. These changes are characteristic of DNA damage caused by UV light. The presence of mutational events at the DNA‐binding surface of the p53 protein may correlate with poor clinical outcome. However, we could not find any statistically significant correlations between p53 status and survival. Only the recurrence‐free interval was significantly shortened in cases with mutations affecting residues of the DNA‐binding surface of the p53 protein. Int. J. Cancer 88:82–86, 2000.

Microvessel densities and microvascular architecture in colorectal carcinomas and their liver metastases: significant correlation of high microvessel densities with better survival.

Aims:  Microvessel densities in cancers have been shown to be a prognostic factor for some types of cancer. For colorectal cancer, however, the situation is far from clear.

Autologous umbilical cord blood mononuclear cell transplantation preserves right ventricular function in a novel model of chronic right ventricular volume overload.

We aimed to evaluate the feasibility and efficacy of autologous umbilical cord blood mononuclear cell (UCMNC) transplantation on right ventricular (RV) function in a novel model of chronic RV volume overload. Four-month-old sheep (n = 20) were randomized into cell (n = 10) and control groups (n = 10). After assessment of baseline RV function by the conductance catheter method, a transannular patch (TAP) was sutured to the right ventricular outflow tract (RVOT). Following infundibulotomy the ring of the pulmonary valve was transected without cardiopulmonary bypass. UCMNC implantation (8.22 ± 6.28 × 107) in the cell group and medium injection in the control group were performed into the RV myocardium around the TAP. UCMNCs were cultured for 2 weeks after fluorescence-activated cell sorting (FACS) analysis for CD34 antigen. Transthoracic echocardiography (TTE) and computed tomography were performed after 6 weeks and 3 months, respectively. RV function was assessed 3 months postoperatively before the hearts were excised for immunohistological examinations. FACS analysis revealed 1.2 ± 0.22% CD34+ cells within the isolated UCMNCs from which AcLDL+ endothelial cells were cultured in vitro. All animals survived surgery. TTE revealed grade II–III pulmonary regurgitation in both groups. Pressure-volume loops under dobutamine stress showed significantly improved RV diastolic function in the cell group (dP/dtmin: p = 0.043; Eed: p = 0.009). CD31 staining indicated a significantly enhanced number of microvessels in the region of UCMNC implantation in the cell group (p < 0.001). No adverse tissue changes were observed. TAP augmentation and pulmonary annulus distortion without cardiopulmonary bypass constitutes a valid large animal model mimicking the surgical repair of tetralogy of Fallot. Our results indicate that the chronically volume-overloaded RV profits from autologous UCMNC implantation by enhanced diastolic properties with a probable underlying mechanism of increased angiogenesis.

HPV DNA and p53 alterations in oropharyngeal carcinomas

We have examined a series of 37 oropharyngeal squamous cell carcinomas for the presence of HPV \611, 16, and 18 DNA by polymerase chain reaction (PCR)/Southern blotting and for p53 alterations by immunohistochemistry and mutation screening with temperature gradient gel electrophoresis (TGGE). HPV sequences were found in a total of 26 of 37 cancers (70.3%), most frequently HPV 16 (\2037) followed by HPV 18 (\1137). Double infections with HPV 16 and 18 were present in 5 tumours. p53 accumulation was detectable immunohistochemically in 21 of 37 carcinomas (56.8%). There were remarkable differences in the distribution of immunoreactive tumour cells in relation to the tumour grade. A mutation screening for p53 by TGGE, directed to the amplified exons 5–8, revealed p53 mutations in 14 of 37 carcinomas (37.8%). Mutations in two different exons were present in 3 tumours, 11 tumours being hit once. Exon 7 was mutated in 6 carcinomas, exons 5 and 8 in 4 cases, and exon 6 in 3 cases. When grouping the tumours with p53 mutation according to their HPV state, HPV-positive cases showed slightly more mutations (\1126) than HPV-negative cases (\311). Only 5 of 37 carcinomas (13.5%) contained neither HPV DNA nor p53 alterations. Our results indicate that high-risk HPV and p53 mutations frequently coexist in oropharyngeal carcinomas, in contrast to genital tumours, notably carcinomas of the cervix uteri. This may reflect different pathways in carcinogenesis in squamous cell epithelium from different sites.

Expression profiling of colorectal carcinomas using tissue microarrays: cell cycle regulatory proteins p21, p27, and p53 as immunohistochemical prognostic markers in univariate and multivariate analysis.

With the rapidly growing understanding of tumor biology, molecular staging of cancer is expected to improve prognostication. This would be particularly important for cancers amenable to adjuvant treatment, such as colorectal carcinomas. To generate data for this, the tissue microarray technique may prove useful. Tissue microarrays were constructed with triplicate cores (0.6 mm diameter) from the invasive margins of a consecutive single-institution series of 184 colorectal carcinomas. Immunostaining for p53, p21, p27, E-cadherin, and β-catenin was scored. Tumor cell proliferation was assessed by mitotic indices and Ki-67 labeling, apoptosis by quantification of apoptotic bodies. Reduced nuclear immunostaining for p21 (<10%) and p27 (≤50%) and reduced membranous expression of E-cadherin were significantly associated with a poorer clinical course by univariate analysis. β-catenin immunostaining had no prognostic impact. Mitotic and apoptotic indices as well as Ki-67 labeling below the median were indicators of poor prognosis. Complete absence of p53 nuclear staining was a significant adverse prognostic factor. By Cox regression, p53 = 0%, p53 = 0%, in combination with p27 ≤50%, the mitotic index and the combined mitotic and apoptotic index added prognostic information to UICC stage. The authors found that growth pattern, lymphohistiocytic response, lymphatic permeation, and venous spread, too, each was a strong prognosticator in addition to UICC stage. The results support that tissue microarrays are a useful tool for screening immunohistochemical markers for prognostic use. An immunopanel of p21, p27, and p53 could be useful for prognostication in colorectal carcinoma in addition to UICC stage.

Persistent carcinoma in cervical cancer screening: non-participation is the most significant cause.

Objective: It was the aim of this study to determine the screening history of all invasive cervical carcinomas between 2004 and 2009 in one of the Federal States of Germany. Study Design: The pooled data sets of all in-state laboratories, corrected and supplemented by data of the State Cancer Registry, were used. The screening histories of all patients, their age and tumor types were collated and analyzed. Results: Of 617 patients with invasive carcinoma of the cervix, 373 (60%) had not had a cervical smear within the past 5 years. In 188 patients (31%), an incomplete screening history was found, whereas only 9% of women had participated regularly. In non-participants, late tumor stages (stage T1B and higher) were predominant and found in 86%. In contrast, in the group with regular screening histories more than half of all cases (54%) were microinvasive carcinomas (stage T1A) with excellent prognosis. Lack of follow-up or refusal of treatment by patients played a minor yet significant role. Conclusions: Non-participation is still by far the most common reason for persistent cases of cervical carcinoma in the German screening program.

Immunohistochemical and glycohistochemical localization of the β-galactoside-binding S-type lectin in human placenta

The localization of the beta-galactoside binding lectin was studied immunohistochemically on acetone-fixed cryostat sections of full-term placental tissue using a biotinylated monoclonal antibody and glycohistochemically applying biotinylated asialofetuin and lactosylated bovine serum albumin. On blots of placental tissue lysates the lectin is recognized by the biotinylated lactosylated bovine serum albumin. The glycoconjugate recognition of the lectin on blots was inhibited in the presence of 0.1 M lactose showing the specificity of the interactions. The anti-lectin monoclonal antibody stained syncytiotrophoblast and trophoblastic cells. Both reagents applied for glycohistochemistry stained syncytiotrophoblast and trophoblastic cells of placental villi and the trophoblastic layer of extraplacental membranes. A strong uniform cytoplasmic staining was characteristic for syncytiotrophoblast and to a lower extent for cytotrophoblastic cells. The localization of the lectin is discussed with respect to a possible immunosuppressive function.