Genomic profiling of late-onset basal cell carcinomas from two brothers with nevoid basal cell carcinoma syndrome.

BACKGROUND: Nevoid basal cell carcinoma syndrome (NBCCS) is an autosomal dominant genetic disorder. It is commonly caused by mutations in PTCH1 and chiefly characterized by multiple basal cell carcinomas (BCCs) developing prior to the age of 30 years. In rare cases, NBCCS presents with a late onset of BCC development. OBJECTIVE: To investigate BCC tumorigenesis in two brothers, who showed characteristic features of NBCCS but developed their first BCCs only after the age of 40 years. Two other siblings did not show signs of NBCCS. RESULTS: We obtained blood samples from four siblings and nine BCCs from the two brothers with NBCCS. Whole exome sequencing and RNA sequencing revealed loss of heterozygosity (LOH) of PTCH1 in eight out of nine tumours that consistently involved the same haplotype on chromosome 9. This haplotype contained a germinal splice site mutation in PTCH1 (NM_001083605:exon9:c.763-6C>A). Analysis of germline DNA confirmed segregation of this mutation with the disease. All BCCs harboured additional somatic loss-of-function (LoF) mutations in the remaining PTCH1 allele which are not typically seen in other cases of NBCCS. This suggests a hypomorphic nature of the germinal PTCH1 mutation in this family. Furthermore, all BCCs had a similar tumour mutational burden compared to BCCs of unrelated NBCCS patients while harbouring a higher number of damaging PTCH1 mutations. CONCLUSIONS: Our data suggest that a sequence of three genetic hits leads to the late development of BCCs in two brothers with NBCCS: a hypomorphic germline mutation, followed by somatic LOH and additional mutations that complete PTCH1 inactivation. These genetic events are in line with the late occurrence of the first BCC and with the higher number of damaging PTCH1 mutations compared to usual cases of NBCCS.

Nuclear iASPP may facilitate prostate cancer progression.

One of the major challenges in prostate cancer (PCa) research is the identification of key players that control the progression of primary cancers to invasive and metastatic disease. The majority of metastatic PCa express wild-type p53, whereas loss of p63 expression, a p53 family member, is a common event. Here we identify inhibitor of apoptosis-stimulating protein of p53 (iASPP), a common cellular regulator of p53 and p63, as an important player of PCa progression. Detailed analysis of the prostate epithelium of iASPP transgenic mice, iASPP(Δ8/Δ8) mice, revealed that iASPP deficiency resulted in a reduction in the number of p63 expressing basal epithelial cells compared with that seen in wild-type mice. Nuclear and cytoplasmic iASPP expression was greater in PCa samples compared with benign epithelium. Importantly nuclear iASPP associated with p53 accumulation in vitro and in vivo. A pair of isogenic primary and metastatic PCa cell lines revealed that nuclear iASPP is enriched in the highly metastatic PCa cells. Nuclear iASPP is often detected in PCa cells located at the invasive leading edge in vivo. Increased iASPP expression associated with metastatic disease and PCa-specific death in a clinical cohort with long-term follow-up. These results suggest that iASPP function is required to maintain the expression of p63 in normal basal prostate epithelium, and nuclear iASPP may inactivate p53 function and facilitate PCa progression. Thus iASPP expression may act as a predictive marker of PCa progression.

Anterior gradient protein 2 (AGR2) is an independent prognostic factor in ovarian high-grade serous carcinoma.

Ovarian high-grade serous carcinoma (HGSC, type 2 ovarian carcinoma) is a poor prognosis cancer with limited therapeutic options. We aimed to investigate the expression pattern and prognostic potential of the metastasis-promoting protein anterior gradient 2 (AGR2) in primary HGSC. Immunohistochemistry was applied to a cohort of 124 primary HGSCs using tissue microarrays. Additionally, in 48 type 1 carcinomas (low-grade serous (LGSC), endometrioid (EC), clear cell (CCC), and mucinous carcinoma (MC)), AGR2 expression was investigated in an exploratory approach. A strong expression of AGR2 was seen in 15 HGSCs (12.1 %) and was significantly linked to shortened overall survival (OS, p = 0.011) and also for progression-free survival (PFS, p = 0.001) in the setting of adjuvant platinum-based chemotherapy (CTX). Multivariate survival analysis including age, stage, and residual tumor after surgery revealed that AGR2 expression was an independent prognostic marker for OS (p = 0.001) and PFS (p = 0.001) in HGSC. In type 1 carcinomas, AGR2 was significantly increased as compared to HGSC (p = 0.001) and was seen in subsets of all histological types, low-grade serous LGSC, EC, CCC, and MC. In particular, strong diffuse staining was seen in LGSC and MC. There was no association between AGR2 and estrogen receptor expression in ovarian type 1 or type 2 carcinomas. AGR2 expression identifies highly aggressive HGSC with a compromised prognosis for which novel therapeutic options are needed. Our data strongly support the further evaluation of AGR2 as a therapeutic target and a potential marker for response to platinum-based CTX in this tumor entity.

Serial NT-proBNP measurements for risk stratification of patients with decompensated heart failure.

PURPOSE: NT-proBNP is an important prognostic predictor in patients with heart failure. However, it is unknown whether a change of NT-proBNP plasma levels in the early phase of decompensation might be of additional prognostic value in patients with acute decompensation of heart failure. METHODS AND RESULTS: NT-proBNP plasma levels of 116 patients with decompensated heart failure from ischemic/non-ischemic origin were measured at baseline and at 12, 24 and 48 h after hospital admission. Baseline levels and changes of plasma levels within the first 48 h were correlated with 30-day mortality. In all patients, NT-proBNP 12 h after admission was highest and superior with respect to the prediction of 30-day mortality compared to plasma levels on admission. In total, 38 patients died within the first 30 days. In these patients absolute NT-proBNP plasma levels were significantly higher and the increase within 12 h after admission was more pronounced compared to survivors (p<0.001). NT-proBNP at 12 h after admission also had the highest predictive value for the 30-day mortality rate in patients with acute myocardial infarction. The increase of NT-proBNP plasma levels within 12 h after admission had the highest predictive value in patients suffering from decompensated heart failure. CONCLUSIONS: NT-proBNP is a powerful marker of 30-day mortality in patients with decompensated heart failure of ischemic and non-ischemic origin. Compared with single baseline measurements, serial measurements of NT-proBNP plasma levels within 12 h after hospital admission may be used to increase the predictive value of NT-proBNP with regard to the early identification of patients who are at high risk of mortality.

Endogenous myoglobin in human breast cancer is a hallmark of luminal cancer phenotype.

BACKGROUND: We aimed to clarify the incidence and the clinicopathological value of non-muscle myoglobin (Mb) in a large cohort of non-invasive and invasive breast cancer cases. METHODS: Matched pairs of breast tissues from 10 patients plus 17 breast cell lines were screened by quantitative PCR for Mb mRNA. In addition, 917 invasive and 155 non-invasive breast cancer cases were analysed by immunohistochemistry for Mb expression and correlated to clinicopathological parameters and basal molecular characteristics including oestrogen receptor-alpha (ERalpha)/progesteron receptor (PR)/HER2, fatty acid synthase (FASN), hypoxia-inducible factor-1alpha (HIF-1alpha), HIF-2alpha, glucose transporter 1 (GLUT1) and carbonic anhydrase IX (CAIX). The spatial relationship of Mb and ERalpha or FASN was followed up by double immunofluorescence. Finally, the effects of estradiol treatment and FASN inhibition on Mb expression in breast cancer cells were analysed. RESULTS: Myoglobin mRNA was found in a subset of breast cancer cell lines; in microdissected tumours Mb transcript was markedly upregulated. In all, 71% of tumours displayed Mb protein expression in significant correlation with a positive hormone receptor status and better prognosis. In silico data mining confirmed higher Mb levels in luminal-type breast cancer. Myoglobin was also correlated to FASN, HIF-2alpha and CAIX, but not to HIF-1alpha or GLUT1, suggesting hypoxia to participate in its regulation. Double immunofluorescence showed a cellular co-expression of ERalpha or FASN and Mb. In addition, Mb levels were modulated on estradiol treatment and FASN inhibition in a cell model. CONCLUSION: We conclude that in breast cancer, Mb is co-expressed with ERalpha and co-regulated by oestrogen signalling and can be considered a hallmark of luminal breast cancer phenotype. This and its possible new role in fatty acid metabolism may have fundamental implications for our understanding of Mb in solid tumours.

[Intrahepatic sarcoma of the follicular dendritic cells]. / Intrahepatisches Sarkom der follikulär-dendritischen Zellen.

We report an intrahepatic sarcoma of the follicular dendritic cells in a 76-year-old woman with a medical history of a hyaline-vascular type of Castleman's disease. We discuss the clinico-pathological findings, the pathogenesis and the differential diagnosis of this rare tumour entity.

[Sialolipoma of the parotid gland]. / Sialolipom der Glandula parotis.

Sialolipoma is a relatively new and rare variant of lipoma of the salivary glands characterized by the combination of classical lipoma morphology with non-neoplastic ductulo-acinary salivary tissue components. Including the presented case, 27 sialolipomas, 14 of them localized in the parotid gland, have been published. We describe the clinical, radiological and pathomorphological characteristics of a parotid sialolipoma in a 43-year-old man.

GOLPH2 protein expression as a novel tissue biomarker for prostate cancer: implications for tissue-based diagnostics.

GOLPH2 is coding the 73-kDa type II Golgi membrane antigen GOLPH2/GP73. Upregulation of GOLPH2 mRNA has been recently reported in expression array analyses of prostate cancer. As GOLPH2 protein expression in prostate tissues is currently unknown, this study aimed at a comprehensive analysis of GOLPH2 protein in benign and malignant prostate lesions. Immunohistochemically detected GOLPH2 protein expression was compared with the basal cell marker p63 and the prostate cancer marker alpha-methylacyl-CoA racemase (AMACR) in 614 radical prostatectomy specimens. GOLPH2 exhibited a perinuclear Golgi-type staining pattern and was preferentially seen in prostatic gland epithelia. Using a semiquantitative staining intensity score, GOLPH2 expression was significantly higher in prostate cancer glands compared with normal glands (P<0.001). GOLPH2 protein was upregulated in 567 of 614 tumours (92.3%) and AMACR in 583 of 614 tumours (95%) (correlation coefficient 0.113, P = 0.005). Importantly, GOLPH2 immunohistochemistry exhibited a lower level of intratumoral heterogeneity (25 vs 45%). Further, GOLPH2 upregulation was detected in 26 of 31 (84%) AMACR-negative prostate cancer cases. These data clearly suggest GOLPH2 as an additional ancillary positive marker for tissue-based diagnosis of prostate cancer.

Do different disorders in cell cycle regulation account for different biological behavior of urothelial and sinonasal inverted papillomas?

OBJECTIVE: It was the aim of this study to assess the expression of selected cell cycle regulation genes in urothelial and sinonasal inverted papillomas (IP). METHODS: Archived surgically resected specimens from 18 urothelial and 19 sinonasal IP were studied immunohistochemically for p16, p53, cyclin D1 and Ki67. Staining results were semiquantified and compared between IP and adjacent control mucosa (CM). RESULTS: p53 expression did not differ between sinonasal and urothelial IP. Although there was a trend of higher p53 expression in IP compared with the adjacent CM in sinonasal and urothelial specimens, this trend failed to be statistically significant. p16 expression was significantly higher in urothelial IP and CM in comparison with their sinonasal counterparts, but did not differ significantly between IP and its adjacent CM either in urothelial or sinonasal specimens. There were no significant differences in the mean scores for cyclin D1 or Ki67. CONCLUSION: The changes in p53 expression seen in both types of IP compared with adjacent CM suggest that sinonasal and urothelial IP may share some common ground in terms of their evolution. Although p16 appears not to be directly involved in the development of sinonasal or urothelial IP, the differing recurrence patterns of sinonasal versus urothelial IP may be attributable in part to different p16 expression.

Histone deacetylases 1, 2 and 3 are highly expressed in prostate cancer and HDAC2 expression is associated with shorter PSA relapse time after radical prostatectomy.

High activity of histone deacetylases (HDACs) causes epigenetic alterations associated with malignant cell behaviour. Consequently, HDAC inhibitors have entered late-phase clinical trials as new antineoplastic drugs. However, little is known about expression and function of specific HDAC isoforms in human tumours including prostate cancer. We investigated the expression of class I HDACs in 192 prostate carcinomas by immunohistochemistry and correlated our findings to clinicopathological parameters including follow-up data. Class I HDAC isoforms were strongly expressed in the majority of the cases (HDAC1: 69.8%, HDAC2: 74%, HDAC3: 94.8%). High rates of HDAC1 and HDAC2 expression were significantly associated with tumour dedifferentiation. Strong expression of all HDACs was accompanied by enhanced tumour cell proliferation. In addition, HDAC2 was an independent prognostic marker in our prostate cancer cohort. In conclusion, we showed that the known effects of HDACs on differentiation and proliferation of cancer cells observed in vitro can also be confirmed in vivo. The class I HDAC isoforms 1, 2 and 3 are differentially expressed in prostate cancer, which might be important for upcoming studies on HDAC inhibitors in this tumour entity. Also, the highly significant prognostic value of HDAC2 clearly deserves further study.

European and US publications in the 50 highest ranking pathology journals from 2000 to 2006.

AIMS: To analyse the contributions of the 15 primary member states of the European Union and selected non-European countries to pathological research between 2000 and 2006. METHODS: Pathological journals were screened using ISI Web of Knowledge database. The number of publications and related impact factors were determined for each country. Relevant socioeconomic indicators were related to the scientific output. Subsequently, results were compared to publications in 10 of the leading biomedical journals. RESULTS: The research output remained generally stable. In Europe, the UK, Germany, France, Italy and Spain ranked top concerning contributions to publications and impact factors in the pathological and leading general biomedical journals. With regard to socioeconomic data, smaller, mainly northern European countries showed a relatively higher efficiency. Of the lager countries, the UK is the most efficient in that respect. The rising economic powers of China and India were consistently in the rear. CONCLUSIONS: Results mirror the leading role of the USA in pathology research but also show the relevance of European scientists. The scientometric approach in this study provides a new fundamental and comparative overview of pathology research in the European Union and the USA which could help to benchmark scientific output among countries.

Co-expression and prognostic value of gross cystic disease fluid protein 15 and mammaglobin in primary breast cancer.

Gross cystic disease fluid protein (GCDFP-15) and mammaglobin are both widely used and accepted markers for epithelia of breast origin. We aimed to evaluate their relation of expression on parallel whole tissue sections in primary breast cancer by immunohistochemistry and also to correlate it with clinico-pathological parameters including patient survival. Primary breast carcinomas from 165 patients with a mean clinical follow-up of 73 months were immunostained using commercially available antibodies against GCDFP-15 and mammaglobin. An immunoreactive score (IRS) was calculated based on the cytoplasmic staining intensity and the number of cells stained. Cytoplasmic expression of GCDFP-15 and mammaglobin was observed in 73.3% and 72.1% of invasive breast carcinomas respectively. 91.8% of breast cancer cases expressed at least one of both markers. Both markers strongly correlated with each other and were significantly associated with lower tumour grading. Additionally, GCDFP-15 negativity was significantly associated with shortened disease-free survival times in univariate and multivariate analyses. We demonstrated the strong correlation of GCDFP-15 and mammaglobin with each other and showed that only very few primary breast cancers are completely negative for both markers. The significantly longer disease free survival times for patients with GCDFP-15 positive tumours clearly warrants further study.

Expression of AGR2 in non small cell lung cancer.

We aimed to evaluate immunohistochemically the expression of the human Anterior Gradient-2 (AGR2), a gene which has recently been proposed as an oncogene for lung carcinoma development, in non small cell lung cancer and to correlate the findings to clinico-pathological data including patient survival. 95 cases of NSCLC were immunostained using a polyclonal AGR2 antibody and statistical analyses were applied to test for prognostic and diagnostic associations. AGR2 was expressed in 66.3% of cases, preferentially adenocarcinomas. There were no relevant associations with clinico-pathological parameters. A prognostic value of AGR2 could not be demonstrated neither in multivariate nor in univariate analyses. Interestingly, this is the first study to demonstrate AGR2 expression in squamous cell carcinomas. Although a prognostic value of AGR2 seems unlikely further studies are warranted to investigate the biological role of AGR2 in NSCLC and its differential expression according to histology.

[Histopathologic quality of prostate core biopsy specimens: comparison of an MR-compatible biopsy needle and a ferromagnetic biopsy needle used for ultrasound-guided prostate biopsy]. / Histopathologische Qualität von Prostatastanzzylindern: Vergleich einer MR-kompatiblen Biopsienadel mit einer im Ultraschall eingesetzten ferromagnetischen Biopsienadel.

PURPOSE: The histopathologic quality of core biopsy specimens obtained via MRI-guided prostate biopsy using a 16G MR-compatible needle was compared to that of biopsies obtained via ultrasound-guided biopsy using a conventional 18G stainless steel biopsy needle. MATERIAL AND METHODS: A retrospective analysis was performed for a total of 247 transrectal prostate biopsy specimens obtained from 32 patients. A total of 117 tissue cores were obtained from 15 patients (PSA of 10.8 ng/ml, age 64 years) who underwent an MRI-guided prostate biopsy using a 16G (1.7 mm) MR-compatible biopsy needle made of titanium alloy. The remaining 130 tissue cores were obtained from 17 patients (PSA of 6.7 ng/ml, age 68 years) who underwent a transrectal ultrasound-guided prostate biopsy using an 18G (1.3 mm) ferromagnetic stainless steel biopsy needle. The length and width of the histologic sections prepared from the tissue cores were measured to calculate the area. The histopathologic quality of the specimens was assessed microscopically using tissue fragmentation, the presence of crush artifacts, and the overall accessibility as criteria. Each of these features was assigned a score from 0 to 3. All 3 features contributed equally to the overall score which ranged from 0 (no tissue) to 9 (optimal quality). RESULTS: The overall quality scores assigned to the biopsies obtained with a 16G MR-compatible needle and an 18G ferromagnetic needle can be considered to be equivalent to a mean difference between patient related median scores of the specimens of - 0.05 (95 % confidence interval [- 0.46; 0.36]) and a given equivalence limit of 1. The MRI biopsies showed more tissue fragmentation (p = 0.001) but fewer crush artifacts (p = 0.022) while the accessibility did not differ significantly between the two needle types (p = 0.064). There was also no significant difference in the calculated areas of the tissue cores (p = 0.236). According to the different calibers of the biopsy needles, the lengths (p = 0.008) and widths (p = 0.000) of the biopsy specimens differed significantly. CONCLUSIONS: The core biopsy specimens obtained with an MR-compatible 16G titanium alloy biopsy needle are of the same histopathologic quality as specimens obtained with a ferromagnetic 18G stainless steel needle.

[Flat epithelial neoplasia and other columnar cell lesions of the breast]. / Flache epitheliale Atypie und andere Zylinderzellläsionen der Brust.

Columnar cell lesions are being increasingly identified in specimens excised for mammographically suspect microcalcifications. The entity of flat epithelial neoplasia remains a challenge for surgical pathologists due to its uncertain biological and clinical significance, inconsistent nomenclature, lack of prognostic data and the often unobtrusive and easily overlooked histological findings. This review aims to summarize our experience and the currently available literature on this topic, and will lead to a better understanding of this lesion. Because of its putative role as a precursor lesion and its many similarities to atypical ductal hyperplasia or ductal carcinoma in situ, differentiation from these lesions on the one hand and from columnar cell lesions without atypia on the other, is of importance and should result in different therapeutic recommendations depending on its presence in excisional or core needle biopsies.

Cytoplasmic overexpression of ALCAM is prognostic of disease progression in breast cancer.

BACKGROUND: Activated leucocyte cell adhesion molecule (ALCAM, CD166) is a cell surface member of the immunoglobulin superfamily. ALCAM expression has prognostic relevance in prostate and colon cancer. OBJECTIVE: To evaluate ALCAM protein expression in breast cancer by immunohistochemistry and to correlate expression levels with clinicopathological data. METHODS: 162 primary breast carcinomas with a mean clinical follow up time of 53 months were immunostained using a monoclonal ALCAM antibody. The staining was evaluated as an immunoreactive score (IRS) and grouped into low v high for both membranous and cytoplasmic staining. RESULTS: Intraductal and invasive carcinomas showed a higher ALCAM expression (median IRS 4 and 6 respectively) than normal breast tissue (IRS 2). In univariate survival analyses a significant association of high cytoplasmic ALCAM expression with shortened patient disease-free survival (mean (SD) five year non-progression rate, 69.4 (4.6)% v 49.4 (11.1)%, p = 0.0142) was found. In multivariate analyses of disease-free survival times, high cytoplasmic ALCAM expression (relative risk (RR) = 2.086, p = 0.026) and nodal status (RR = 2.246, p = 0.035) were significantly associated with earlier disease progression, whereas tumour grading (RR = 1.6, p = 0.052) was of borderline significance. CONCLUSIONS: The data suggest that strong cytoplasmic ALCAM expression in primary breast cancer, as detected by immunohistochemistry, might be a new marker for a more aggressive breast cancer biology.

Expression of human Kallikrein 14 (KLK14) in breast cancer is associated with higher tumour grades and positive nodal status.

Human kallikrein 14 (KLK14) is a steroid hormone-regulated member of the tissue kallikrein family of serine proteases, for which a prognostic and diagnostic value in breast cancer has been suggested. To further characterise the value of KLK14 as a breast tumour marker, we have carefully analysed KLK14 expression in normal breast tissue and breast cancer both on the RNA level by real-time RT-PCR (n = 39), and on the protein level (n = 127) using a KLK14-specific antibody for immunohistochemistry. We correlated KLK14 protein expression data with available clinico-pathological parameters (mean follow-up time was 55 months) including patient prognosis. KLK14 RNA expression as quantified by real-time RT-PCR was significantly more abundant in breast tumours compared to normal breast tissue (P = 0.027), an issue that had not been clarified recently. Concordantly with the RNA data, cytoplasmic KLK14 protein expression was significantly higher in invasive breast carcinomas compared to normal breast tissues (P = 0.003). Furthermore, KLK14 protein expression was associated with higher tumour grade (P = 0.041) and positive nodal status (P = 0.045) but was not significantly associated with shortened disease-free or overall patient survival time in univariate analyses. We conclude that KLK14 is clearly overexpressed in breast cancer in comparison to normal breast tissues and is positively associated with conventional parameters of tumour aggressiveness, but due to a missing association with survival times, the use of KLK14 immunohistochemistry as a prognostic marker in breast cancer is questionable.

Ergonomic assessment of the static stress confronted by surgeons during laparoscopic cholecystectomy.

BACKGROUND: Only a few reports on static strain in the spine, neck, and head of the surgeon are available, describing it as "distinctly harmful." The aim of this study was to objectively prove the static burden during laparoscopic operations. For this, new industrial software called PCMAN was used, capable of measuring and comparing the postures of the surgeon at different monitor placements. METHODS: Two simultaneous and synchronized video recordings of laparoscopic cholecystectomies (LC) were done using miniDV digital camcorders with the cameras standing at a 90 degrees angle to each other. Twenty operations were performed using two different placements of the monitor. In 10 cases, the monitor was placed at the patient's head in the center, and in 10 cases at the left side of the patient. Using the time codes of the recordings, different steps of the operation were identified, and the duration of these measured in seconds. Very characteristic, longer lasting postures were imported to and analyzed with the software. Results of the different setups were compared to each other, and to an "ideal" comfort posture. RESULTS: During the intermediate steps of the operations the rate of static phases is significantly higher. Measuring the typical postures of these phases the trunk and head are significantly more rotated and bent than in comfort positions. When the monitor was at the side of the patient facing the surgeon, results were closer to the comfort posture. CONCLUSIONS: It was proven that surgeons are confronted by significant static burden during LC. The software used was able to evaluate objectively the static posture of the surgeon during series of LC. Results also confirmed that the position of monitors significantly influences the surgeon's posture. Best setups for the whole team can be achieved by adjustable multiple monitor systems.