Expression of microRNA 210 associates with poor survival and age of tumor onset of soft-tissue sarcoma patients.

Expression of microRNAs can affect age of tumor onset and prognosis of cancer patients. However, nothing is known about the effects of microRNAs on altered age of cancer onset and disease-specific survival of soft-tissue sarcoma (STS) patients. The levels of miR-210, also known as hypoxia-regulated microRNA, were analyzed by quantitative real-time (RT)-PCR in the tumors of 78 STS patients. The patients were stratified according to their microRNA levels with low, intermediate and high expression levels and the association of microRNA expression and patients' survival was analyzed using multivariate Cox's regression hazard analyses. A significant correlation between an intermediate miR-210 expression and disease-specific death of STS patients [relative risk (RR) = 3.19; p = 0.018] was observed compared with patients with high expression levels in their tumors. Interestingly, the association between an intermediate expression of miR-210 and a poor prognosis was only significant in female STS patients (RR = 11.28; p = 0.010), but not observed in male individuals. Furthermore, the expression of miR-210 showed a significant association with the age of tumor onset in a gender-specific manner. Specifically, male patients with an intermediate expression of miR-210 associated with a 9.6-year later age of tumor onset (p = 0.017) compared with males with a low expression of miR-210 in their tumors. However, no significant differences in the female patients were observed. This study provides the first evidence of a correlation of expression levels of a single microRNA (miR-210) with the prognosis and age of tumor onset in a gender-specific manner in STS patients.

The tumor suppressor RASSF10 is upregulated upon contact inhibition and frequently epigenetically silenced in cancer.

The Ras association domain family (RASSF) comprises a group of tumor suppressors that are frequently epigenetically inactivated in various tumor entities and linked to apoptosis, cell cycle control and microtubule stability. In this work, we concentrated on the newly identified putative tumor suppressor RASSF10. Methylation analysis reveals RASSF10 promoter hypermethylation in lung cancer, head and neck (HN) cancer, sarcoma and pancreatic cancer. An increase in RASSF10 methylation from normal tissues, primary tumors to cancer cell lines was observed. Methylation was reversed by 5-aza-2'-deoxycytidine treatment leading to reexpression of RASSF10. We further show that overexpression of RASSF10 suppresses colony formation in cancer cell lines. In addition, RASSF10 is upregulated by cell-cell contact and regulated on promoter level as well as endogenously by forskolin, protein kinase A (PKA) and activator Protein 1 (AP-1), linking RASSF10 to the cAMP signaling pathway. Knockdown of the AP-1 member JunD interfered with contact inhibition induced RASSF10 expression. In summary, we found RASSF10 to be epigenetically inactivated by hypermethylation of its CpG island promoter in lung, HN, sarcoma and pancreatic cancer. Furthermore, our novel findings suggest that tumor suppressor RASSF10 is upregulated by PKA and JunD signaling upon contact inhibition and that RASSF10 suppresses growth of cancer cells.

Co-detection of members of the urokinase plasminogen activator system in tumour tissue and serum correlates with a poor prognosis for soft-tissue sarcoma patients.

BACKGROUND: The urokinase plasminogen activator (uPA) system is one of the best-investigated protease systems, both under physiological and pathological conditions, including various types of cancer. However, effects of co-expression of members of the uPA system in soft-tissue sarcoma (STS) patients at the protein level in both tumour tissue and serum have not been investigated yet. METHODS: We examined 82 STS patients for protein levels of uPA, PAI-1and uPAR in tumour tissue and serum by ELISA. RESULTS: A significant correlation between high antigen levels of uPA, PAI-1 or uPAR in tumour tissue, and of uPAR in serum, with poor outcome of STS patients was found for the first time. Most strikingly, we observed an additive effect of combined uPA, PAI-1 or uPAR levels in tumour tissue extracts with uPAR levels in serum on patients' prognosis. High uPA/uPAR, PAI-1/uPAR and uPAR/uPAR antigen levels in tumour tissue/serum were associated with a 5.9-fold, 5.8-fold and 6.2-fold increased risk of tumour-related death (P=0.003, 0.001 and 0.002, respectively) compared with those patients who displayed low levels of the respective marker combination. CONCLUSION: As expression of members of the uPA system in tumour tissue and serum is additively correlated with prognosis of STS patients, our results suggest that combinations of these biomarkers can identify STS patients with a higher risk of tumour-related death.

Interferon-gamma release assay in the ascites: Early hint for diagnosis of abdominal tuberculosis.

We report on a 20-year-old woman with abdominal tuberculosis.Standard microbiological examination of ascites showed no acid-fast bacilli (AFB), and analysis for the Mycobacterium tuberculosis (MTB)-complex DNA by PCR was negative. However,the interferon-gamma release assay (IGRA) of the ascites was positive after specific stimulation with mycobacterial antigens(ESAT-6/CFP-10/TB7.7[p4]), indicating an infection with MTB.The diagnosis of tuberculosis was later confirmed by histology, MTB culture, and PCR analysis of MTB-complex DNA in tissue samples taken during laparoscopy. Thus, the IGRA of ascites may guide the decision to start active treatment for tuberculosis.

The stem cell-associated Hiwi gene in human adenocarcinoma of the pancreas: expression and risk of tumour-related death.

Piwi proteins and their interaction with piRNAs have rapidly emerged as important contributors to gene regulation, indicating their crucial function in germline and stem cell development. However, data on the Hiwi 1 (Hiwi) gene, one of the four human Piwi homologues, are still scarce. Therefore, we investigated the Hiwi mRNA expression in microdissected PDAC tissues from patients with ductal adenocarcinoma of the pancreas (PDAC) by quantitative real-time PCR and the protein expression by immunohistochemistry. Elevated levels of Hiwi mRNA transcripts were measured in 40 out of 56 tissues and a positive immunostaining of Hiwi was detected in tumours of 21 out of 78 patients. There was no general impact of elevated Hiwi mRNA transcript levels or protein expression on survival, as tested by multivariate Cox regression and Kaplan-Meier analysis. However, men showed a significantly increased risk for tumour-related death in case of down- or upregulated expression of Hiwi mRNA (relative risk (RR)=2.78; P=0.034). In summary, we report the first analysis of Hiwi expression in PDAC and its impact on prognosis. We suggest that alterations in mRNA expression of Hiwi can increase the risk of tumour-related death in male PDAC patients.

Stem cell-associated genes are extremely poor prognostic factors for soft-tissue sarcoma patients.

Cancer stem cells can play an important role in tumorigenesis and tumor progression. However, it is still difficult to detect and isolate cancer stem cells. An alternative approach is to analyse stem cell-associated gene expression. We investigated the coexpression of three stem cell-associated genes, Hiwi, hTERT and survivin, by quantitative real-time-PCR in 104 primary soft-tissue sarcomas (STS). Multivariate Cox's proportional hazards regression analyses allowed correlating gene expression with overall survival for STS patients. Coexpression of all three stem cell-associated genes resulted in a significantly increased risk of tumor-related death. Importantly, tumors of patients with the poorest prognosis were of all four tumor stages, suggesting that their risk is based upon coexpression of stem cell-associated genes rather than on tumor stage.

Expression of the stem cell self-renewal gene Hiwi and risk of tumour-related death in patients with soft-tissue sarcoma.

Self-renewal is considered as a common property of stem cells. Dysregulation of stem cell self-renewal is likely a requirement for the development of cancer. Hiwi, the human Piwi gene, encodes a protein responsible for stem cell self-renewal. In this study, we investigated the expression of Hiwi at the RNA level by real-time quantitative PCR in 65 primary soft-tissue sarcomas (STS) and ascertained its impact on prognosis for STS patients. In a multivariate Cox's proportional hazards regression model, we found that an increased expression of Hiwi mRNA is a significant negative prognostic factor for patients with STS (P=0.017; relative risk 4.6, 95% confidence interval (CI) 1.3-16.1) compared to medium expression of Hiwi transcript. However, a low expression of Hiwi transcript is correlated with a 2.4-fold (CI 0.7-8.0) increased risk, but this effect was not significant (P=0.17). Altogether, high-level expression of Hiwi mRNA identifies STS patients at high risk of tumour-related death. This is the first report showing a correlation between expression of a gene involved in stem cell self-renewal and prognosis of cancer patients.

Histopathological outcome of 597 isolated soft tissue tumors suspected of soft tissue sarcoma: a single-center 12-year experience.

BACKGROUND: The aim of this present report was to analyze the patients referred to us with the presumptive diagnosis of soft tissue sarcoma (STS). METHODS: We reviewed all patients referred to us with suspected soft tissue sarcoma (STS) of the extremities or trunk over a 12-year period. RESULTS: We treated 597 patients with soft tissue tumors. Open biopsy revealed soft tissue sarcoma in 318 cases, benign mesenchymal tumor in 124 cases and isolated metastases (ISTM) from carcinomas in 98 patients; other pathologies were found in 57 patients. The primary carcinomas were lung cancer in 26 patients, breast cancer in 19 patients, renal carcinoma in 16 patients, carcinoma of the esophagus in 12 patients, colonic carcinoma in 5 patients, thyroid gland cancer in 6 patients, and in 14 patients carcinoma of unknown primary was diagnosed. CONCLUSIONS: In our collective with soft tissue tumor, 50% of the patients had the diagnosis of soft tissue sarcoma, 20% presented with a metastasis of carcinoma and 20% had a benign tumor. Referring to our results, in patients with the presumptive diagnosis of soft tissue sarcomas, soft tissue metastasis of a primary carcinoma was unexpectedly common, indicating that greater consideration should be given to this differential diagnosis.

[Wound infection and infection-promoting factors in breast cancer surgery -- a prospective multicenter study on quality control]. / Wundinfektionen und sie begünstigende Faktoren im Rahmen der Mammakarzinomchirurgie -- prospektive Multizenterstudie zur Qualitätssicherung.

INTRODUCTION: The quality of treatment of cancer of the female breast is reflected not only in such parameters as local recurrence rate and survival times, but also in the development of surgical complications. Within the framework of a study investigating the performance and quality assurance in surgical treatment of breast cancer, therefore, the wound infection rate (WIR) and factors influencing it were analysed in a large patient population. METHODS: In the period between 1.1.2000 and 31.12.2000, 84 surgical departments participated in a prospective multicenter study to investigate primary surgery for breast cancer. A total of 1 416 patients were recruited to the study, the organization and conduction of which was in the hands of the former surgical department 1 of the University of Leipzig under the patronage of the East German Working Group for Performance and Quality Control in Surgery in cooperation with the An Institute for Quality Control in Operative Medicine of the Otto-von-Guericke University in Magdeburg. In addition to parameters characterizing patients, tumors and diagnostic work-up, we also analysed the surgical treatment and its possible complications with the aid of a questionnaire. The definition of wound infection was based on the criteria of the "Hospital Infection Control Practice Advisory Committee". RESULTS: The overall WIR was 4.5 % (n = 65). 21 (32 %) of the wound infections (WI) were diagnosed exclusively on a clinical basis without establishing the responsible pathogens. In 44 (68 %) of the WI, a search for the pathogen was undertaken which in 3 cases (7 %) was negative, and in 41 cases (93 %) positive. 118 (8.3 %) of the patients received perioperative antibiotic cover. The following parameters were found to have a significant influence on WIR: local drainage, blood transfusion, the time lapse between biopsy and definitive surgery, and the size of the primary tumor. DISCUSSION: Some of the above factors (transfusion, time lapse, drainage) can be influenced by the therapist. The wound infection rate is a marker for treatment quality.

[Differential diagnosis of soft tissue sarcomas]. / Differentialdiagnose von Weichteilsarkomen Zehnjahresanalyse eines Zentrums mit 490 unter Weichteilsarkomverdacht zugewiesenen Patienten.

INTRODUCTION: Soft tissue sarcoma often goes undetected. PATIENTS AND METHODS: Over a 10-year period, the patients referred to us with a soft tissue tumor (STT) of the extremities and wall of the trunk were analyzed retrospectively. The aim of the present study was to investigate the differential diagnoses, the number of incompletely operated STS, and local recurrences together with their percentage fluctuations. RESULTS: A total of 490 patients with an STT were referred to our department, and of these patients 55% were diagnosed with an STS. In addition to STS, the differential diagnoses for STT included 2% lymphomas, 18% isolated carcinoma metastases, 18% benign mesenchymal tumors, 5% inflammatory processes, and 2% old hematomas. Only 45% of the STS had not undergone previous surgery. Of these, 15% had been incompletely resected, while 39% of the STS patients were admitted with a local recurrence. Within the 10-year period, referrals with STT and STS remained relatively constant, but referrals of patients with incompletely resected or recurrent STS doubled in the last 2 years under observation. DISCUSSION: In view of the numerous differential diagnoses of an STT, both the possibility of an STS and also carcinoma manifestations in the soft tissues should receive more attention. With the aim of reducing the relatively high number of STS re-resections and local recurrences, the treatment of patients with suspicious STT should be reserved for a specialized center.

Expression of alternatively and aberrantly spliced transcripts of the MDM2 mRNA is not tumor-specific.

The MDM2 proto-oncogene encodes a 90-kDa protein that binds to and inactivates the tumor suppressor p53. Several reports describe the presence of different alternatively, as well as, aberrantly spliced transcripts of the MDM2 mRNA in a variety of human cancers that have lost the ability to bind p53. Due to the transforming ability of at least some of the isoforms it has been suggested that they might contribute to tumorigenesis. Here we show that shorter MDM2 transcripts are also widely expressed in normal tissues, including lung and renal tissue, and in lymphocytes. Alteration in MDM2 RNA transcripts were found in the majority of the samples. Although we cannot exclude that alterations in MDM2 preferentially occur during cancer development, our data rather indicate that in this context the commonly observed transcript variants may also possess a normal physiological function.

[Prospective multicenter study for quality management of breast cancer surgery]. / Prospektive Multizenterstudie zur Qualitätssicherung der Chirurgie des Mammakarzinoms.

Over the last 10 years a dramatic decrease became apparent in primary treatment of breast cancer in general surgical departments. A prospective 1-year observational study involving 84 surgical departments was carried out to describe the current therapeutic situation. A total of 1,416 patients undergoing primary surgical treatment for mammary carcinoma were recorded, and their data evaluated. 68.9% of the carcinomas were treated in departments with an annual case load for this disease of more than 20 operations, with 50% of them being operated on in 8 departments with a case load of 40-100 procedures per year. 94.4% of the carcinomas were confirmed histologically, and in 91% of the patients surgery was performed in curative intention. The rate of breast-preserving procedures was 40%, and breast amputations accounted for 60%. An analysis of the data allowed an evaluation of this specific patient group in the surgical departments. Deficits in terms of management quality are identified.

Expression of cathepsin B, D and L protein in juvenile idiopathic arthritis.

Juvenile idiopathic arthritis (JIA) is the most common childhood autoimmune rheumatic disease and like rheumatoid arthritis (RA), it is characterized by inflammation and the progressive destruction of joints. In RA, cathepsins as proteinases play a major role in destroying synovial tissue and cartilage matrix. So far no data on cathepsin expression in pannus tissue of HA patients exist. The aim of this study was to characterize the expression levels of cathepsins B, D, H, and L in HA and to compare them with those in RA. Synovectomy tissue from 16 HA and 12 RA patients was investigated for cathepsin expression levels by Western blot analysis. Expression of cathepsins B, D and L was on comparable levels in the synovectomy tissue of HA and RA patients. The following graduation of expression was determined: cathepsin D > cathepsin L > cathepsin B. Cathepsin H was neither found to be expressed in HA nor in RA patients. The expression levels of cathepsins in pannus tissue showed no clear difference between patients with systemic JIA and patients with monoarticular JIA. In summary, the comparable expression of cathepsins B, D and L in RA and JIA synovectomy tissue suggests that they may play a similarly important role in destroying synovial tissue and cartilage matrix in the course of HA and RA.

[Port catheter fractures in 361 implanted port systems. Analysis of the causes--possible solutions--review of the literature]. / Portkatheterbrüche bei 361 implantierten Portsystemen. Ursachenanalyse--Lösungsmöglichkeiten--Literaturübersicht.

INTRODUCTION: The use of a venous port-catheter-system is known as a relatively safe implant. Besides infections, the breakage of PIPS is the most common reason for explanation before term. The purpose of this study is to analyse port-related complications and to show ways of preventing them. METHOD: Between 1.1.1994 and 31.12.1999, 391 PIPS were implanted in the V. subclavia with the Seldinger technique at Surgical Clinic 1 of the University of Leipzig. Subsequently, 311 of them were followed up until 31.12.2000, with a mean observation time of 45 months. RESULTS: We registered 48 complications altogether (15.4% of 311), 21(6.7%) of which occurred immediately after implantation (up to 30 days postoperatively). These could be divided either into wound-healing disorders/pulmonary distress (4.5%, n = 14) or complications concerning the catheter systems (2.3%, n = 7). Long-term complications after 31 days were evident in 27 patients (8.7%), due either to infections (4.5%, n = 13) or catheter-associated problems (4.5%, n = 14). Catheter lesions occurred in nine cases (2.9% out of 311) at the point of entry into the musculus pectoralis, i.e., where the catheter had to change direction. Typically these were lengthways tears caused by the catheter. We observed one full breakage without dislocation, and two dislocated catheter fragments in the systemic circulation. We consider the change of direction to be responsible for wear on the silicon catheter. During implantation, extreme change of direction of the catheter should be avoided because this is where breakage happens. Catheter implantation by means of exposure of the vena basilica in the infraclavicular triangle is the method of choice.

[Inflammatory pseudotumor of the ileopsoas muscle with femoral paralysis caused by massive metal abrasion of a hip endoprosthesis]. / Entzündlicher Pseudotumor im M. ileopsoas mit Femoralisparese durch massiven Metallabrieb einer Hüftendoprothese.

Case report of a 62-year-old patient with a presumed loosening of a hip endoprosthesis after 10 years and a planned replacement. In addition, the patient suffered pain in the thigh and had paresis of the femoral nerve. A CAT-Scan substantiated the diagnosis either of a suppurating or a neoplastic tumour in the left iliac foss. The wide excision revealed a rare inflammatory tumour in the left ileopsoas muscle due to the excessive abrasion of the metal hip endoprosthesis.

[Intraoperative electrophysiological monitoring of the recurrent laryngeal nerve in thyroid gland surgery--a prospective study]. / Anwendung des Neuromonitoring des N. laryngeus recurrens in der Schilddrüsenchirurgie - eine prospektive Studie.

INTRODUCTION: In the last few years the use of intraoperative electrophysiological monitoring of the recurrent laryngeal nerve (RLN) in thyroid gland surgery has become more and more important. PATIENTS AND METHOD: In a prospective study 223 nerves at risk in 116 patients were monitored with the Neurosign(R)100 (Fa. Magstim Ltd., UK). We used intramuscular needle electrodes inserted into the vocal muscle through the conic ligament. Practicability, complications, acceptance and predictive value of the method were documented. Recurrent nerve palsy rate and complications were compared with a control group operated upon without monitoring. RESULTS: The intraoperative delay using this method was on average 8.9 minutes. There were problems with monitoring equipment avoiding use in 6.4 %. In 2 cases (1.7 %) an accidental lesion of endotracheal tube cuff was found related to malpositioning of the needle and in 7.7 % a hematoma of the vocal cords was observed. 73.3 % of the surgeons accepted the method to identify and control the nerve integrity. False-positive and false-negative signals may occur. In cases of a final real stimulus response a regular vocal cord motility was found in 95 %. If a nerve conduction block was noted an immobility of ipsilateral vocal cord was diagnosed postoperatively in 50 %. There was no decrease in transient recurrent palsy rate using monitoring (10.7 % vs. 9.6 % without monitoring) but in permanent paralysis (1.8 % vs. 3.0 %). CONCLUSIONS: It may be concluded that intraoperative electrophysiological monitoring of the RLN is a simple and accepted method with low complications reducing the incidence of permanent RLN palsy rate. We found the monitoring especially useful for operations of recurrent goiter and carcinomas of the thyroid gland as well as for learning thyroid gland surgery.

[A giant malignant tumour of the skull-Case report and differential diagnosis]. / Ein gigantischer maligner Tumor der Kalotte - Fallbericht und Differentialdiagnose.

The case of a 72-year-old woman with a high-partially located tumor grown within a half year to a magnitude of 8.5 x 11 x 11 cm is reported. The patient remembered a mastectomy and axillary lymphadenectomy followed by chemotherapy and radiation 8 years ago. Therefore we assumed a skeletal metastasis of a breast cancer. After wide excision, an unusual morphology was found, allowing only a classification as a pleomorphic sarcoma. Searching for the pathohistological evaluation of the former breast tumor, a cystosarcoma phylloides malignum could be found out. The tumor described here can be identified as a metastasis of this rare neoplasm.

Gains of 12q are the most frequent genomic imbalances in adult fibrosarcoma and are correlated with a poor outcome.

Comparative genomic hybridization was used to analyze 41 adult fibrosarcomas from 34 patients. Thirty-one patients showed in their tumors DNA sequence copy number changes (mean 11, range 3-25). The minimal common regions for the most frequent gains were narrowed down to 12q21 (18 cases); 12q14-q15 and 14q22 (16 cases each); 4q22, 7q31, and 14q23-q24 (15 cases each); and 4q21, 4q23-q24, 8q22, and 12q22 (14 cases each). Twenty-five high-level amplifications were observed in 12 samples. 12q21 and 18p were affected three times each; and 1p21, 4q31.3, 7p21, 12q14-q15, Xp22.1-p22.2, and Xq22-q23 two times each. Losses were less frequent than gains. Early stages of adult fibrosarcomas were characterized by frequent gains of chromosomes 2, 4q, and 14q, whereas gains of chromosomes 7 and 8q were associated with progression. Gains of 12q were frequent in all of the developmental steps of this soft-tissue sarcoma. By investigation of several tumors of the same patient, a number of corresponding changes were always detected. Adult fibrosarcomas from patients who died during the observation time showed statistically significant more frequent gains of 8q, 12q, 13q, and 15q compared to the fibrosarcomas of patients who are alive. Gains and high-level amplifications of 12q14-q22, which were the most frequent genomic imbalances, partly reflected an MDM2 amplification, indicating the importance of this region in the tumorigenesis of sarcomas. In adult fibrosarcomas, a gain of 12q22 correlated significantly (P = 0.028) with a poor overall survival rate.

[Nonpalpable carcinoma of the breast - diagnosed by vacuum core breast biopsy and surgical management]. / Nonpalpable Mammakarzinome - Vakuumstanzbioptische Sicherung und chirurgisches Management -

The wire-localized extirpation is the "gold standard" for the examination of nonpalpable lesions suspicious of malignancy. Less invasive techniques were introduced in the last years, offering also a high diagnostic reliability, e. g. stereotactic core needle biopsy, the "advanced breast biopsy" and the vacuum core biopsy. Based on an analysis of 40 vacuum core breast biopsies and the following interventions in the case of carcinoma recommendations for the management of the nonpalpable breast carcinoma diagnosed by vacuum core biopsy should be developed. In 12 patients (33 %) carcinomas were found necessitating further operations. These were 92 % pTis or pT1pN0M0-carcinomas and only in one case an occult pT2pN1M0-carcinoma. We recommend a short interval between core biopsy and operation, a preoperative localization of the clips e. g. the residual microcalcification, and the controlled placement of the hooked wire that should also be performed at the Mammotome(R) using the same way to the tumor. Furthermore it is necessary to excise the core biopsy localization channel en bloc together with a wide tumour excision. An intraoperative histological examination of the specimen should be performed to confirm tumour-free excision borders. For this, the position of specimen should be marked by a thread and a specimen radiography should be made for the orientation of the pathologist and for documentation. A long-term follow-up of these patients under study conditions should be considered. Patients with benign diagnosis, not undergoing general anesthesia and operation with the consequences for later radiological evaluation, mostly profit from vacuum core breast biopsy. For patients with carcinoma the costs of the perioperative management increase. This should have consequences for the quality assurance of this method.

Increased survivin transcript levels: an independent negative predictor of survival in soft tissue sarcoma patients.

Survivin, a recently identified inhibitor of apoptosis protein (IAP), is expressed in diverse embryonic tissues and in various human cancers. We have investigated the quantitative expression of survivin mRNA by a sensitive TaqMan-based RT-PCR assay in tissue samples from 94 patients with soft tissue sarcomas (STS). Survivin transcript levels were measured and normalized to GAPDH transcripts. By using a multivariate Cox regression analysis, we found an inverse correlation between the level of survivin mRNA (ratio >2 zmol survivin/amol GAPDH) and the rate of overall survival (p = 0.009, RR = 2.7). Survivin transcript variants as detected by qualitative RT-PCR analysis were revealed in 36 of 56 STS patients (64%). Only survivin DeltaEx3 and/or full-length survivin variants but not survivin 2B were identified. Our results suggest that a higher level of survivin mRNA is an independent predictor of survival for STS patients.