DNA flow-cytometric analysis in colorectal cancer: A comparison of metastasizing and non-metastasizing tumours

The most common cause of death in patients with colorectal cancer is metastatic liver disease. In order to identify patients at a high risk of developing hepatic secondaries from colorectal cancers, DNA content was measured in metastasizing colorectal primaries (Group I, n= 32) as well as in their subsequently resected liver secondaries and in sections of non-metastasizing colorectal cancers (Group II, n= 25). A modified interpretation system involving both a DNA index and percentage of cycling cells (those in S and G2 + M phases) was developed. DNA content was measured in paraffin-embedded sections by flow cytometry using internal controls (human peripheral blood mononuclear cells) and non-malignant tissue controls (19 patients with diverticular disease). In Group I there were significantly more tumours with both abnormal ploidy (aneuploid or abnormal tetraploid peak) and > 15% cycling cells compared with Group II (Chi-squared; P= 0.034). The combination of abnormal ploidy and > 15% cycling cells was superior to Dukes’ classification for identifying metastasizing tumours (Logistic Regression; P= 0.047). However, it was not possible to discriminate between the two groups using either DNA ploidy or the percentage of cycling cells alone. The metastasizing colorectal cancers exhibited similar DNA ploidy characteristics and had a similar percentage of cycling cells compared with their liver metastases. These results suggest that tumour DNA ploidy plus the percentage of cycling cells may predict the development of liver metastases and thus survival in patients with colorectal cancer.     

ATRX encodes a novel member of the SNF2 family of proteins: mutations point to a common mechanism underlying the ATR-X syndrome

It was shown recently that mutations of the ATRX gene give rise to a severe, X-linked form of syndromal mental retardation associated with alpha thalassaemia (ATR-X syndrome). In this study, we have characterised the full-length cDNA and predicted structure of the ATRX protein. Comparative analysis shows that it is an entirely new member of the SNF2 subgroup of a superfamily of proteins with similar ATPase and helicase domains. ATRX probably acts as a regulator of gene expression. Definition of its genomic structure enabled us to identify four novel splicing defects by screening 52 affected individuals. Correlation between these and previously identified mutations with variations in the ATR-X phenotype provides insights into the pathophysiology of this disease and the normal role of the ATRX protein in vivo.      

Recurrence of Cutaneous Necrosis in an Infant with Probable Catastrophic Antiphospholipid Syndrome

Abstract: We present the case of a 3‐month‐old child with probable catastrophic antiphospholipid syndrome who, after initial successful management with immunomodulary therapies including rituximab, experienced a cutaneous relapse. This rare event was successfully re‐treated with repeated administration of rituximab, supporting its role in the control of this disorder. Dermatologic manifestations may be the main clinical presentation of antiphospholipid syndrome, a possible underdiagnosed but potentially fatal pathology.     

Human mesenchymal stem cells at the single-cell level: simultaneous seven-colour immunofluorescence

Extracellular, intracellular or surface proteins can be used as putative markers to characterize human mesenchymal stem cells (hMSC). However, these markers are also expressed by other cell types and primary cell pools reveal considerable heterogeneity. Therefore, the simultaneous detection of several markers on a single cell appears to be an attractive approach to identify hMSC. Here we demonstrate the specific distinction of human MSC from human osteoblasts via seven-colour fluorescence on the single cell level with simultaneous marker detection of CD44, CD105/endoglin, CD106/VCAM-1, collagen-IV, fibronectin, actin and DAPI nuclear staining. We performed spectral image acquisition using a Sagnac-type interferometer. Subsequent linear unmixing allowed for decomposition of each pixel in its spectral components. Our approach reveals a typical expression profile of the adherent singular cells, allowing the specific distinction between hMSC and osteoblasts on the single cell level.     

Interventionelle Therapieverfahren bei akuter nekrotisierender Pankreatitis

The clinical course of acute pancreatitis is variable. Severe pancreatitis is observed in up to 20% of cases and is associated with high mortality rates of up to 40%. The most serious complication is the infection of the (peri-)pancreatic necroses. The therapeutic goal is debridement of the infected material. Whereas surgical methods still represent the gold standard, minimally invasive interventional approaches are gaining importance. This article reviews the different interventional procedures, particularly percutaneous, CT-guided drainage and necrosectomy. Furthermore, an overview of published studies about interventional therapy in patients with acute necrotizing pancreatitis is given.     

Precursor B-cell lymphoblastic lymphoma in childhood and adolescence: clinical features, treatment, and results in trials NHL-BFM 86 and 90

BACKGROUND: Precursor B-cell lymphoblastic lymphoma (PBLL) is a rare subtype of childhood non-Hodgkin lymphoma (NHL). The purpose of our study was to investigate frequency and clinicopathological features of PBLL in children and to test prospectively the efficacy of an ALL-type therapy for treatment of these patients. PROCEDURE: From October, 1986, to March, 1995, 1,075 patients up to 18 years of age suffering from all kinds of NHL were registered in the two consecutive multicenter studies NHL-BFM 86 and 90. Of these, 27 patients were diagnosed with PBLL. Twenty-one PBLL patients were treated according to a BFM-ALL-type protocol: an eight-drug induction over 9 weeks was followed by an 8-week consolidation including methotrexate 5 g/m(2) x4. Patients in stages I and II continued with maintenance up to a total therapy duration of 24 months, whereas patients in stages III and IV received an additional eight-drug intensification and cranial radiotherapy (12 Gy for prophylaxis) after consolidation. Six PBLL patients were treated according to the BFM-protocol for B-NHL, stratified according to stage and tumor load and consisiting of two to six 5-day courses of chemotherapy. RESULTS: The median age of the 27 patients with PBLL (18 boys, 9 girls) was 6.2 (range 0.7-15) years. Stages (St. Jude) were: I (n = 3), II (n = 7), III (n = 9), and IV (n = 8). Twenty-one PBLL patients had nodal disease, 6 patients had subcutaneous manifestations, and 8 patients had bone marrow disease (<25% blasts). All patients achieved remission. With a median follow-up time of 4. 25 years, the estimated probability for event-free survival (pEFS) at 10 years for the total group was 0.73 (SE 0.10). Five patients (2, 1, 1, and 1 patients at stages I, II, III, and IV, respectively) relapsed: 2 of 21 patients who were treated according to the ALL strategy and 3 of 6 who were treated according to the B-NHL-protocol. CONCLUSIONS: PBLL accounts for 2.5% of childhood NHL. An ALL-type therapy strategy appears to be superior to a short-pulse B-NHL protocol.     

Epitopic mapping of structural and nonstructural Aleutian disease virus proteins

Common antigenic properties for p85 and p75 but a different antigenic character for p71 Aleutian disease virus (ADV) proteins were demonstrated by Western blot analysis with monoclonal antibodies. It was shown that four hybridomas (ADV-Hy 47, 66, 77 and 84) with specific reactivity for structural proteins p85 and p75 also recognized p25 but not the p71, nonstructural, protein. In turn, the monoclonal antibody ADV-Hy 2 recognized the p71 protein only. For further studies of their antigenic properties, the ADV proteins were subjected to enzymatic or chemical cleavage. The derived peptide fragments were analyzed by epitopic mapping. Depending on the cleavage reagent and monoclonal antibody applied, specific peptide maps were revealed. The maps of p85 and p75 were very similar, indicating that both proteins shared an extensive antigenic relationship. After cleavage with alpha-chymotrypsin and N-chlorosuccinimide and by using the ADV-Hy 84 monoclonal antibody, unique peptide fragments were identified with p85 which had no counterparts in p75 fragments.