Threshold values for stress radiographs in unstable knees after total knee arthroplasty.

PURPOSE: The primary aim of this study was to investigate the potential benefit of stress radiographs for diagnosis of unstable total knee arthroplasty (TKA) and to identify clinically relevant cut-off values to differentiate between unstable and stable TKAs. METHODS: Data of 40 patients with 49 cruciate retaining (CR) TKA who underwent stress radiographs as part of the diagnostic algorithm in a painful knee clinic were prospectively collected. Anterior and posterior stress radiographs were done in 90° and 15° flexion, varus-valgus stress radiographs in 0° and 30° knee flexion. Knee laxity was measured in mm and degrees by two independent observers using standardized landmarks. Intra- and inter-observer single measure intraclass correlations were between 0.92 to 1 and 0.89 to 1, respectively. For evaluation and investigation of the potential cut-off values, two groups of patients with and without revision surgery due to instability were compared. Radiographic measures of standardized z values according to the group without revision due to instability were used to calculate average and maximum laxity z-scores. RESULTS: Knees undergoing revision TKA due to instability showed significantly (p < 0.001) lower (KSS) pain/function scores (94 ± 6.3, range 80-100; control group: 112 ± 19.2, range 80-148) and total KSS scores when compared to the control group. The laxity values of patients with instability were significantly higher in terms of mean values (p < 0.01) when compared to the control group. The maximum laxity z-score showed the strongest difference between the groups (R2 = 0.26, p < 0.001). The following cut-off values indicating need of revision due to instability were established: in 90° (15°) flexion-anterior translation 5.2 mm (22.4 mm), posterior translation 16.6 mm (13.2 mm); varus stress in 0° (20°-30°) flexion-inlay gap 5.2 mm (6.1 mm) or joint angle 6.1° (6.8°); valgus stress in 0° (20°-30°) flexion-inlay gap 4.6 mm (5.7 mm) or joint angle 5.2° (7.1°). CONCLUSION: Standardized stress radiographs are helpful tools for diagnosis of instability after TKA. The established cut-off values help to guide decision making in this challenging group of patients. However, laxity values should not be considered as the only criteria for diagnosis of unstable TKA. LEVEL OF EVIDENCE: IV.

[Magnetic resonance arthrography]. / MR-Arthrographien.

Magnetic resonance (MR) arthrography is a sensitive examination technique. In this institute the majority of MR arthrography examinations are carried out on the shoulder (57 %), followed by hand (24 %) and hip joints (16 %). The use of MR arthrography is superior to native MR imaging (MRI) particularly for detection of partial thickness tendon tears of the shoulder joint. Nevertheless, this technique is invasive, time-consuming and may be uncomfortable for patients. This article summarizes the indications, risks and contraindications of MR arthrography, highlights the puncture technique and illustrates possible pitfalls.

Ongoing MRI remodeling 3-7 years after collagen meniscus implantation in stable knees.

PURPOSE: The purpose of the present study was to evaluate the clinical and radiological 3-7 years outcomes of patients who underwent collagen meniscus implantation in stable or stabilized knees. It was the hypothesis that using the collagen meniscus (CMI) good clinical 3-7 years outcomes with low pain levels are achieved. METHODS: Thirty-nine patients (male:female = 30:9, mean age 34 ± 10 years) underwent arthroscopic CMI after subtotal medial (n = 32) or lateral meniscectomy (n = 7). A 7-mm CMI was performed due to prophylactic (n = 25) or therapeutic indication (n = 14). IKDC score, Tegner score preinjury, preoperatively and at follow-up, Lysholm score and visual analogue scale for pain and satisfaction (follow-up rate 90%) were assessed. MRI scans were analyzed according to the Genovese criteria (n = 19). Implant failure was defined as infection or mechanical failure of the device. The minimum follow-up time was 36 months (range 36-84 months). RESULTS: The mean VAS satisfaction preoperatively and at follow-up was 4.0 ± 0 and 1.6 ± 1.0. The mean VAS pain was 4.3 ± 3.2 preoperatively and at last follow-up 2.1 ± 1.7. The median Tegner score preinjury was 7 (range 3-10), it decreased preoperatively to median 3.5 (range 1-8) and nearly reached the preinjury level at last follow-up 6 (range 3-10). The mean Lysholm score before surgery was 66 ± 20 and 91 ± 8 at last follow-up. Seven patients (38.9%) had a normal total IKDC score (A), 10 patients were nearly normal (B) and 1 patient slightly abnormal (C). In MRI the CMI was entirely resorbed in 4 patients (21%) and partially resorbed in 15 (79%). In 4 patients (21%) the CMI was isointense, in 14 (74%) slightly hyperintense and in 1 (5%) highly hyperintense. Ten patients (53%) showed marked signs of bone marrow edema. In 13 patients (68%) an extrusion of the meniscus > 3 mm at last follow-up was found. CONCLUSIONS: Meniscal substitution with the CMI showed good to excellent clinical 3-7 results. The CMI shows an ongoing remodelling with decreased signal intensity and decreased size. However, as meniscus extrusion remained at the same level and bone marrow edema decreased from 1 year to longer term follow-up, it appears that the remodeling comes to an end at about 5 years after CMI. LEVEL OF EVIDENCE: IV.

Cost-effectiveness of nivolumab in the treatment of head and neck cancer.

BACKGROUND: Until recently, no second-line treatment for recurrent and/or metastatic head and neck squamous cell cancer (r/mHNSCC) was able to improve overall survival (OS). Nivolumab has become a promising treatment for r/mHNSCC. The CheckMate-141 trial showed that nivolumab improves OS compared to investigator's choice (IC) (cetuximab, methotrexate, docetaxel). Treatment with immune checkpoint inhibitors is however expensive. The aim of this analysis was to assess the cost-effectiveness of nivolumab as second-line treatment for r/mHNSCC in Switzerland. METHODS: Based on the CheckMate-141 trial, we constructed a Markov model comparing nivolumab to IC, including follow-up data up to 24 months. We assessed costs for treatments from the perspective of the Swiss health system with a 60 months' time horizon. PD-L1 and p16 testing were considered in scenarios. Incremental cost-effectiveness ratios (ICER) were compared to an informal willingness-to-pay of CHF (Swiss Francs) 100,000 per QALY gained. RESULTS: For the base case we estimated an incremental effectiveness of 0.35 QALYs and incremental costs of CHF 35,562 with nivolumab, resulting in an ICER of CHF 102,957 per QALY gained. Most influential drivers for the ICER were the price of nivolumab and the progressive disease state utility weights. In 45.5% of probabilistic sensitivity analysis simulations nivolumab was estimated below 100,000 CHF/QALY. Reducing the price of nivolumab according to a consented payback by 4.75%, resulted in an ICER of CHF 98,325/QALY gained. CONCLUSIONS: At current prices nivolumab has an ICER of around CHF 100,000 per QALY gained in the second line treatment of r/mHNSCC patients in Switzerland.

[Assessment of ligament and tendon injuries of the elbow using magnetic resonance imaging]. / Band- und Sehnenverletzungen des Ellenbogens in der Magnetresonanztomographie.

An overview about the normal anatomy and frequent pathologies of tendons and ligaments of the elbow using MRI will be provided. The unique joint configuration and articulation, as well as passive ligamentous and active muscle structures as well as the unique configuration of the articulating components of the elbow contribute to joint stabilization. MRI is an essential imaging modality in patients with ligamentous injuries including the sequelae of joint dislocation as well as chronic pathologies such as long-standing and refractory tendinopathies. Ideally, when reporting MRI of the elbow, the joint is separated into its four compartments, anterior, posterior, medial and lateral, allowing precise, comprehensive and structured reporting.

Calcium pyrophosphate deposition disease: a frequent finding in patients with long-standing erosive gout.

OBJECTIVE: To characterize patients with both monosodium urate (MSU) and calcium pyrophosphate (CPP) crystals in their synovial fluid (SF). METHOD: Forty-nine gout patients with acute arthritis were included. Those patients with MSU crystals only in their SF were compared to those patients with both MSU and CPP crystals in their SF. RESULTS: A total of 36 out of 49 patients (73.5%) had only MSU crystals, whereas 13 out of 49 (26.5%) had both MSU and CPP crystals in their SF. Co-deposition of CPP crystals was associated with long-standing gout disease (p = 0.022), kidney dysfunction (p = 0.024), and erosive arthritis (p = 0.049), but not with age. CONCLUSION: Long-standing gout may be a risk factor for CPP deposition disease, and the frequency of CPP co-deposition may be higher than expected.

Systematic Radiation Dose Reduction in Cervical Spine CT of Human Cadaveric Specimens: How Low Can We Go?

BACKGROUND AND PURPOSE: While the use of cervical spine CT in trauma settings has increased, the balance between image quality and dose reduction remains a concern. The purpose of our study was to compare the image quality of CT of the cervical spine of cadaveric specimens at different radiation dose levels. MATERIALS AND METHODS: The cervical spine of 4 human cadavers (mean body mass index; 30.5 ± 5.2 kg/m2; range, 24-36 kg/m2) was examined using different reference tube current-time products (45, 75, 105, 135, 150, 165, 195, 275, 355 mAs) and a tube voltage of 120 kV(peak). Data were reconstructed with filtered back-projection and iterative reconstruction. Qualitative image noise and morphologic characteristics of bony structures were quantified on a Likert scale. Quantitative image noise was measured. Statistics included analysis of variance and the Tukey test. RESULTS: Compared with filtered back-projection, iterative reconstruction provided significantly lower qualitative (mean noise score: iterative reconstruction = 2.10/filtered back-projection = 2.18; P = .003) and quantitative (mean SD of Hounsfield units in air: iterative reconstruction = 30.2/filtered back-projection = 51.8; P < .001) image noise. Image noise increased as the radiation dose decreased. Qualitative image noise at levels C1-4 was rated as either "no noise" or as "acceptable noise." Any shoulder position was at level C5 and caused more artifacts at lower levels. When we analyzed all spinal levels, scores for morphologic characteristics revealed no significant differences between 105 and 355 mAs (P = .555), but they were worse in scans at 75 mAs (P = .025). CONCLUSIONS: Clinically acceptable image quality of cervical spine CTs for evaluation of bony structures of cadaveric specimens with different body habitus can be achieved with a reference mAs of 105 at 120 kVp with iterative reconstruction. Pull-down of shoulders during acquisition could improve image quality but may not be feasible in trauma patients with unknown injuries.

The Genovese grading scale is not reliable for MR assessment of collagen meniscus implants.

BACKGROUND: The purpose of the study was to evaluate the intra- and inter-observer reliabilities of the Genovese grading on MRI in patients after collagen meniscus substitution. METHODS: 84 MRI images of 74 consecutive patients who underwent partial meniscus substitution using collagen meniscus implant (CMI) were assessed. MRIs were evaluated using the Genovese grading system. Furthermore, meniscal extrusion was assessed. Two observers performed the grading twice, blinded to each other and to the previous results, with a six weeks interval. The inter- and intra-observer reliabilities were assessed using kappa and weighted kappa values. RESULTS: The criterion "morphology/size" showed a weighted kappa value inter-observer reliability of 0.069 (rater 1)/0.352 (rater 2) and intra-observer reliability of 0.170 (1st rating)/0.582 (2nd rating). The criterion "signal intensity" showed a weighted kappa value inter-observer reliability of 0.175/0.284 and intra-observer reliability of 0.294/0.458. The criterion "cartilage lesions" showed a kappa value inter-observer reliability of 0.091/0.525 and intra-observer reliability of 0.409/0.413. The criterion "bone marrow edema" showed a kappa value inter-observer reliability of 0.667/0.808 and intra-observer reliability of 0.702/0.715. The criterion "cartilage lesions" showed a kappa value inter-observer reliability of 0.091/0.525 and intra-observer reliability of 0.409/0.413. Regarding meniscal extrusion kappa values for the inter-observer reliability were 0.625/0.940 and 0.625/0.889 for intra-observer reliability. CONCLUSIONS: Three of the four Genovese grading items showed only slight to moderate inter- and intra-observer reliabilities in evaluating CMI on MRI. Hence, such grading results need to be considered with all due care. Only the criteria "bone marrow edema" and "meniscal extrusion" showed a good agreement for both inter- and intra-observer reliabilities.

Nanoparticulate CpG immunotherapy in RAO-affected horses: phase I and IIa study.

BACKGROUND: Recurrent airway obstruction (RAO), an asthma-like disease, is 1 of the most common allergic diseases in horses in the northern hemisphere. Hypersensitivity reactions to environmental antigens cause an allergic inflammatory response in the equine airways. Cytosine-phosphate-guanosine-oligodeoxynucleotides (CpG-ODN) are known to direct the immune system toward a Th1-pathway, and away from the pro-allergic Th2-line (Th2/Th1-shift). Gelatin nanoparticles (GNPs) are biocompatible and biodegradable immunological inert drug delivery systems that protect CpG-ODN against nuclease degeneration. Preliminary studies on the inhalation of GNP-bound CpG-ODN in RAO-affected horses have shown promising results. OBJECTIVES: The aim of this study was to evaluate the clinical and immunological effects of GNP-bound CpG-ODN in a double-blinded, placebo-controlled, prospective, randomized clinical trial and to verify a sustained effect post-treatment. ANIMALS AND METHODS: Twenty-four RAO-affected horses received 1 inhalation every 2 days for 5 consecutive administrations. Horses were examined for clinical, endoscopic, cytological, and blood biochemical variables before the inhalation regimen (I), immediately afterwards (II), and 4 weeks post-treatment (III). RESULTS: At time points I and II, administration of treatment rather than placebo corresponded to a statistically significant decrease in respiratory effort, nasal discharge, tracheal secretion, and viscosity, AaDO2 and neutrophil percentage, and an increase in arterial oxygen pressure. CONCLUSION AND CLINICAL IMPORTANCE: Administration of a GNP-bound CpG-ODN formulation caused a potent and persistent effect on allergic and inflammatory-induced clinical variables in RAO-affected horses. This treatment, therefore, provides an innovative, promising, and well-tolerated strategy beyond conventional symptomatic long-term therapy and could serve as a model for asthma treatment in humans.

One-year clinical and MR imaging outcome after partial meniscal replacement in stabilized knees using a collagen meniscus implant.

PURPOSE: The purpose of the study was to evaluate the clinical and radiological outcomes after medial/lateral collagen meniscus substitution (CMI) at 12 months postoperatively. METHODS: Sixty-seven patients (m:f = 47:20, mean age 36 ± 10 years) underwent arthroscopic CMI after previous subtotal medial (n = 55) or lateral meniscectomy (n = 12) due to persistent joint line pain (n = 25) or to prophylactic reasons (n = 42). Clinical follow-up consisted of IKDC score, Tegner score, Lysholm score, and visual analog scale for pain and satisfaction (preinjury, preoperatively, and 12 months postoperatively; follow-up rate 90 %). MRI scans were analyzed according to the Genovese criteria. RESULTS: Nineteen patients (29 %) showed a normal (A), 35 nearly normal (B), 5 abnormal (C), and 1 patient severely abnormal total IKDC score (D). The median Tegner preinjury score was 7 (range 2-10) and at follow-up 6 (range 2-10). The mean Lysholm score before surgery was 68 ± 20 and 93 ± 9 at follow-up. Preoperatively, the mean VAS pain was 4.4 ± 3.1 and 2.0 ± 1.0 at follow-up. Clinical failure of the CMI occurred in 3 patients (n = 1 infection, n = 1 failure of the implant, n = 1 chronic synovitis). On MRI, the CMI was completely resorbed in 3 patients (5 %), partially resorbed in 55 (92 %), and entirely preserved in 3 (5 %) patients. In 5 patients (8 %) the CMI was isointense, in 54 (90 %) slightly and 1 (2 %) highly hyperintense. 43 (72 %) patients showed an extrusion of the CMI implant of more than 3 mm. CONCLUSIONS: Significant pain relief and functional improvement throughout all scores at 1 year was noted. The CMI undergoes significant remodeling, degradation, resorption, and extrusion in most of the patients. No difference in outcomes between the medial and lateral CMI was observed. LEVEL OF EVIDENCE: Prospective therapeutic study, Level IV.

KRAS mutation analysis on low percentage of colon cancer cells: the importance of quality assurance.

KRAS mutation testing is mandatory for patients with metastatic colorectal cancer who are eligible for treatment with an epidermal growth factor receptor targeting agent, since tumors with a mutation are not sensitive to the drug. Several methods for mutation testing are in use and the need for external quality assurance has been demonstrated. An often little addressed but important issue in external quality assurance schemes is a low percentage of tumor cells in the test samples, where the analytical sensitivity of most tests becomes critical. Using artificial samples based on a mixture of cell lines with known mutation status of the KRAS gene, we assessed the reliability of a series of commonly used methods (Sanger sequencing, high resolution melting, pyrosequencing, and amplification refractory mutation system-polymerase chain reaction) on samples with 0, 2.5, 5, 10, and 15 % mutated cells. Nine laboratories throughout Europe participated and submitted a total of ten data sets. The limit of detection of each method differed, ranging from >15-5 % tumor cells. All methods showed a decreasing correct mutation call rate proportionally with decreasing percentage of tumor cells. Our findings indicate that laboratories and clinicians need to be aware of the decrease in correct mutation call rate proportionally with decreasing percentage of tumor cells and that external quality assurance schemes need to address the issue of low tumor cell percentage in the test samples.

FISH analysis for diagnostic evaluation of challenging melanocytic lesions.

The differential diagnosis of malignant melanomas and atypical melanocytic nevi is still a diagnostic challenge. The currently accepted morphologic criteria show substantial interobserver variability, likewise immunohistochemical studies are often not able to discriminate these lesions reliably. Techniques that support diagnostic accuracy are of the greatest importance considering the growing incidence of malignant melanomas and their increase in younger patients. In this study we analyzed the feasibility of fluorescence in situ hybridization (FISH) analysis for the discrimination of malignant and benign melanocytic tumors. A panel of DNA probes was used to detect chromosomal aberrations of chromosomes 6 and 11. On a series of 5 clearly malignant and benign melanocytic tumors we confirmed the applicability of the test. Then we focused on examination of ambiguous melanocytic lesions, where atypical cells are often difficult to relocalize in the 4',6-Diamidino-2-phenylindol (DAPI)-fluorescence stain. FISH analyses were conducted on destained H&E-stained slides. By comparison of the DAPI-image with photos taken from the H&E stain, unambiguous assignment of the FISH results to the conspicuous groups of cells was possible. The results of FISH analysis were consistent with the conventional diagnosis in 11 of 14 small ambiguous lesions. Of the remaining 3 cases, 2 showed FISH-results close to the cut-off level. Comparison of FISH results on thin and thick sections revealed that the cut-off values have to be adapted for 2 microm destained sections. In conclusion, FISH analysis is a useful and applicable tool for assessment of even smallest melanocytic neoplasms, although there will remain unclear cases that cannot be solved even after additional FISH evaluation.

EGFR fluorescence in situ hybridisation assay: guidelines for application to non-small-cell lung cancer.

There is a need for predictive biomarkers that identify non-small-cell lung cancer (NSCLC) patients most likely to respond to epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI) treatment. There are numerous potential candidates, although none has been proven in prospective clinical trials. The EGFR gene copy number evaluated by fluorescence in situ hybridisation (FISH) has been highlighted as one of the most effective markers for sensitivity to EGFR TKIs in large phase III, randomised placebo-controlled trials and has been used in clinical settings to assist physicians in defining the therapeutic regimen. The EGFR FISH assay has technical challenges and it is critical that detailed guidelines are provided to help clinical laboratories in performing and interpreting the test. Excellent assay reproducibility and portability rates among laboratories are crucial to guarantee that accurate clinical decisions can be made for patients with NSCLC. This article discusses the consensus outcomes of a global workshop convened to discuss key technical issues and standardise reading strategies for the EGFR FISH assay of NSCLC tumour tissue.

Intraductal carcinoma is the precursor of carcinoma ex pleomorphic adenoma and is often associated with dysfunctional p53.

AIMS: Although intraductal carcinoma has been demonstrated in intracapsular carcinoma ex pleomorphic adenoma (CEPA), the morphological and genetic stages of transformation of pleomorphic adenoma (PA) to CEPA are not fully understood. The aim of this study was to investigate the morphology of intracapsular CEPA. METHODS AND RESULTS: The largest series of intracapsular CEPA studied was subject to immunohistochemical double-staining to detect p53 protein and cellular proliferation in different types of cell combined with mutational analysis of the p53 gene in laser-microdissected material. Intraductal carcinoma with high-grade cellular atypia and frequent accumulation of p53 protein was found in 15/19 cases. Purely intraductal carcinoma was found in eight cases. Mutation of p53 was found in 7/19 cases, of which it was found in intraductal carcinoma in 5/15 cases. CONCLUSIONS: The frequent demonstration of intraductal carcinoma indicates that this preinvasive lesion is likely to be a constant feature in the malignant transformation of PA to CEPA. It appears to be a feature of CEPA developing from both primary and recurrent PA. The combined immunohistochemical and genetic data show that 14/19 cases of CEPA and 11/15 cases with intraductal carcinoma showed genetic or morphological evidence of dysfunctional p53, indicating that this is an early event in malignant transformation.

Microsatellite instability and loss of heterozygosity in prostatic carcinomas: comparison of primary tumors, and of corresponding recurrences after androgen-deprivation therapy and lymph-node metastases.

BACKGROUND: The molecular mechanisms leading to prostate cancer progression are poorly understood. In particular, those changes which are responsible for androgen-independent growth and metastatic spread in prostate cancer are an issue of current investigations. METHODS: To gain more insight into these processes, paired microdissected samples from both untreated, locally advanced primary tumors (n = 20) and recurrences (n = 20) after conventional androgen-deprivation therapy (ADT) were analyzed retrospectively for microsatellite instability (MSI) and loss of heterozygosity (LOH) at nine loci on chromosomes 8, 18, and X by polymerase chain reaction. In parallel, 12 prostatic carcinomas treated by radical prostatectomy and nine corresponding lymph-node metastases were analyzed in the same way. RESULTS: The group treated with ADT showed a total of 10 MSI in 7 of the primary tumors (35%): 4 of these (20%) at one locus, and 3 of these (15%) at two loci. In the recurrences, MSI was observed in 4 cases (20%): 3 of these at one locus (15%), and 1 of these (5%) at two loci. LOH was found in 8 cases (40%) before as well as after ADT. In the radically resected carcinomas, MSI could be detected at two chromosomal loci in one of the primary tumors (8%) and in one of the metastases (11%); LOH was found in 2 primaries (16%) and 3 metastases (33%). CONCLUSIONS: Although MSI can be found in advanced prostatic carcinomas, it apparently does not play a major role in the progression of prostate cancer regarding androgen-independent growth or lymphogenous spread.

Microsatellite analysis in serous tumors of the ovary.

Thirty-four serous ovarian neoplasms were analyzed for microsatellite instability (MIN) and loss of heterozygosity (LOH) at 20 chromosomal loci of eight different chromosomes, including the map positions of the six known mismatch repair genes. Twelve of the 20 (60%) serous ovarian tumors of low malignant potential (LMP) and 13 of 15 (87%) samples of serous ovarian carcinomas, taken from 14 patients, showed LOH at one or more of the analyzed microsatellite loci. In serous carcinomas, LOH of the dinucleotide repeat D7S521 was most frequent. Six of 20 (30%) LMP tumors were also affected by MIN at more than one locus, whereas in the carcinomas MIN was found only sporadically (p < 0.025). No correlation between increased occurrence of MIN and LOH at the chromosomal loci of the known mismatch repair genes were discovered for these LMP tumors. Although our observation regarding LOH frequency in serous LMP tumors and serous carcinomas is compatible with the hypothesis that serous LMP tumors might represent precursor lesions of invasive carcinomas, the results concerning the occurrence of MIN suggest that the mechanisms of tumorigenesis of some tumors of LMP are distinct from those in invasive serous carcinomas.

In serous ovarian neoplasms the frequency of Ki-ras mutations correlates with their malignant potential.

We analysed 44 tissue samples from serous ovarian neoplasms of different malignant potential for Ki-ras mutations by denaturing gradient gel electrophoresis (DGGE) and direct sequencing after microdissection. Point mutations at codon 12 were found in 7 of 20 tumours of low malignant potential (LMP) (35%) and in 2 of 6 well-differentiated carcinomas (33%). In contrast, no mutations were detected in the 11 poorly differentiated ovarian carcinoma samples or in the 7 serous cystadenomas. The frequency of Ki-ras mutations in serous ovarian tumours seems to correlate with the malignant potential of the neoplasms. The data favour the hypothesis of a de novo development of poorly differentiated ovarian carcinomas and do not support an evolution from LMP tumours or well-differentiated carcinomas.

MDM-2 oncoprotein overexpression, p53 gene mutation, and VEGF up-regulation in angiosarcomas.

The endothelium is one of the largest cellular compartments of the human body and has a high proliferative potential. However, angiosarcomas are among the rarest malignancies. Despite this interesting contradiction, data on growth and angiogenesis control mechanisms of angiosarcomas are scarce. In this study of 19 angiosarcomas and 10 benign vascular control lesions we investigated the sequence and expression of the p53 tumor suppressor gene and the expression of the mdm-2 proto-oncogene, which is a negative regulator of p53 activity and of the vascular endothelial growth factor (VEGF), whose expression, among other factors, is regulated by the p53/MDM-2 pathway. Ten sarcomas (53%) exhibited clear nuclear p53 protein accumulation. Two of these cases revealed mutations in the sequence-specific DNA binding domain of the p53 gene. Thirteen angiosarcomas (68%) showed an increased amount of MDM-2 protein. Elevated expression of p53 and MDM-2 protein correlated with increased VEGF expression, which was found in nearly 80% of the angiosarcoma cases. Negative or clearly lower immunostaining was obtained in cases from the benign control collective. Only one case of a juvenile hemangioma reached the cutoff value of p53 positivity coincidentally with high VEGF expression. Our data suggest that the p53/ MDM-2 pathway is impaired in about two-thirds (14/ 19) of the angiosarcomas. This may be a key event in the pathogenesis of human angiosarcomas. The increased VEGF expression observed supports this hypothesis.

Improvement of nonradioactive DNA in situ hybridization.

With the introduction of microwave pretreatment, the quality of nonradioactive in situ hybridization (NISH) using DNA probes on formalin fixed tissue has significantly improved. Even after microwave treatment, however, there are cases where NISH results remain unsatisfactory. Therefore, we tried to improve NISH by testing other buffer systems as alternatives to the citrate buffer that is routinely applied during microwave pretreatment. By using buffer systems originally designed for immunohistochemistry, we significantly improved our NISH results. Difficult tissue samples were more accessible to NISH using these alternative buffer systems and made the quantitative evaluation easier. These results may also be of interest for combined applications of NISH and immunohistochemistry.

[HELLP syndrome: recurrence in 4 consecutive pregnancies]. / HELLP-Syndrom: Wiederholtes Auftreten in vier aufeinanderfolgenden Schwangerschaften.

Recurrence in Four Consecutive Pregnancies in a Patient: We report on the recurrence of HELLP-syndrome in four consecutive pregnancies in a patient. The observation of recurrence of this rare disorder (0.2-0.8% of all pregnancies) in an otherwise healthy patient points to an immunological or genetic predisposition for the development of HELLP-syndrome. At the moment there is no possibility to identify these patients with a high recurrence risk for HELLP-syndrome before the onset of pregnancy.