Modulation of malignant B cell activation and apoptosis by bcl-2 antisense ODN and immunostimulatory CpG ODN

Inhibition of bcl-2 expression by antisense oligodeoxynucleotides (ODN) might render bcl-2 overexpressing malignant B cells more susceptible to chemotherapy. ODN containing unmethylated CG dinucleotides (CpG) are known to activate B cells. We studied the effects of two bcl-2 antisense ODN, with (G3139) or without CG dinucleotides (NOV 2009) within the sequence, and the effects of a nonantisense, CpG-containing ODN (ODN 2006) on activation and apoptosis of malignant B cell lines and primary B-CLL cells. Without cationic lipids, no antisense-mediated inhibition of bcl-2 synthesis was achieved with G3139 and NOV 2009. Instead, G3139, but not NOV 2009, induced similar changes as ODN 2006 in proliferation, expression of costimulatory and antigen-presenting molecules, as well as in bcl-2 and bcl-xL levels of primary B-CLL cells. G3139 and ODN 2006 inhibited in vitro, spontaneous apoptosis in B-CLL cells of patients with high serum thymidine kinase activity (s-TK, marker for proliferative activity of malignant B cells), whereas in patients with low s-TK activity, apoptosis was induced. In conclusion, our results suggest that modulation of malignant B cell apoptosis by G3139 depends on its immunostimulatory properties rather than on antisense-mediated reduction of bcl-2 expression. Immunostimulatory CpG ODN may have a therapeutic potential in patients with B-CLL, especially those with low s-TK activity.     

Interferon-producing cells: on the front line in immune responses against pathogens

Interferon-producing cells (IPC) constitute a small population of leukocytes that secrete high levels of type I interferons in response to viruses. Although human IPC have been known for almost two decades, murine IPC have been identified only recently. Furthermore, it has been shown that IPC correspond to the enigmatic 'plasmacytoid cells' identified in human lymph nodes during infections. These breakthroughs have brought IPC to the attention of immunologists for their role in innate immunity and in shaping T cell responses. Here we review recent progress and outstanding questions in the field.     

Prognostic value of [<Superscript>18</Superscript>F]FDG-PET imaging in small cell lung cancer

Positron emission tomography (PET) utilizing fluorine-18 fluorodeoxyglucose (FDG) has been used in the evaluation of non-small cell lung cancer (NSCLC). Recently its use in the staging of small cell lung cancer (SCLC) has been reported. However, the prognostic value of FDG-PET imaging in SCLC has not been studied. We performed a retrospective analysis to assess this, with the following hypotheses: (1) PET-positive patients would have a less favorable prognosis than PET-negative patients and (2) a high standardized uptake value (SUV) would be associated with a poor prognosis. Retrospective review of a mixed population of treated and untreated patients imaged between 1995 and 2000 was performed. Results of 62 scans in 46 patients were analyzed. There were 8 untreated and 38 treated patients. Findings were correlated with pathology, computed tomography/magnetic resonance imaging and clinical data. The sensitivity of PET scanning was 100% with pathological correlation. The prognostic value of a positive PET study was determined. Overall survival in PET-positive cases was significantly worse than that in PET-negative cases ( P=0.0108). Correlation of SUV(max) with survival showed a significant negative correlation ( P=0.0021). In the eight untreated patients, scans were strongly positive and in all cases the scan results concurred with the final clinical stage assigned on the basis of conventional methods. We conclude that FDG-PET imaging provides prognostic information in treated patients. A positive study and a high SUV(max) are significantly associated with poor survival. Additionally, FDG-PET may be helpful in staging and follow-up.     

Bcl-2 and bax expression in advanced non-small cell lung cancer: lack of correlation with chemotherapy response or survival in patients treated with docetaxel plus vinorelbine

Surgical series of non-small cell lung cancer (NSCLC) pathologic samples have shown that the expression of the proteins bcl-2 and bax, which regulate cell death, may have prognostic implications. Laboratory data also suggests that these proteins may impact chemotherapy response. In order to determine the rate of bcl-2 and bax expression in advanced NSCLC and assess the impact on chemotherapy response and patient survival, we performed immunohistochemistry on biopsy samples from patients enrolled in a phase I/II trial of vinorelbine plus docetaxel. We chose to study the pathology of patients in this specific trial because both docetaxel and vinorelbine phosphorylate bcl-2 and we hypothesized that this mechanism may affect clinical outcome. The goal of this study was to determine the feasibility of this analysis, and to observe any differences in response rate or survival based on bcl-2 or bax staining results. Unstained slides from paraffin blocks were obtained for 31 patients (55%) on the phase I/II trial. The patient characteristics for this subgroup did not differ significantly from the entire cohort of patients on the trials. Bcl-2 staining was positive in 5/31 samples (16%, 95% CI 3-29%) and bax was positive in 19/28 samples (68%, 95% CI 51-85%). Bcl-2 and bax staining did not correlate with response (p = 0.65 and 1.00 respectively, Fisher's exact test), or survival (by Kaplan-Meier curves). In conclusion, bcl-2 and bax expression was similar in this population with advanced NSCLC to previously reported results for early stage disease, but did not predict response to vinorelbine plus docetaxel in this series.     

Lack of sialidase activity in Candida albicans and Candida glabrata

Because several micro-organisms having close contact to animal hosts and man produce sialidase (EC 3.2.1.18) as a tool for adhesion and invasion, we investigated two Candida species for the presence of this enzyme. Two sensitive assays, a fluorometric test with 4-methylumbelliferyl-alpha-D-N-acetylneuraminic acid and a radiometric test with 3H-labelled sialyllactitol as sialidase substrates, were applied to detect sialidase activity. None of 40 Candida albicans and 10 C. glabrata strains grown in three different media exhibited sialidase activity, but the positive control Ophiostoma stenoceras produced sialidase under comparable conditions. Our surprising negative findings are divergent from an earlier positive report, which may be due to strain selection or bacterial contamination. These results indicate that sialidase is probably of no relevance in cutaneous or mucosal candidoses.     

Der Spiraltrokar – ein neues Trokarkonzept

Zusammenfassung. Einleitung: Vorgestellt wird ein neuartiges Trokarprinzip für die minimal-invasive Chirurgie. Es handelt sich um eine Spirale, deren Wendel sukzessive nach zentral hin an St?rke zunimmt. Dadurch entsteht ein flie?ender übergang von Spirale über Schraube bis hin zum Stab. Der Au?endurchmesser bleibt dabei gleich. Der Spiraltrokar wird wie ein Korkenzieher durch die Bauchdecke geschraubt. Der prim?re Punktionskanal verl?uft daher als Spirale. Im Weiteren wird der Kanal dilatiert und zentriert, so dass der stabf?rmige Anteil des Trokars mit der darüber geführten Arbeitshülse durch die Bauchdecke geschoben werden kann. Das Trokardesign weist folgende Vorteile auf: Der Trokar kann kontrolliert durch die Bauchdecke geschraubt werden ohne dass dabei axialer Druck angewendet werden muss. Dadurch wird die Gefahr von Organverletzungen minimiert. Da die verschiedenen Gewebeschichten an unterschiedlichen Stellen punktiert werden, k?nnen sie sich nach Entfernung der Arbeitshülse wieder gegenseitig verschlie?en. Der verbleibende Punktionskanal ist sehr klein und das Risiko der Entstehung von Trokarkanalhernien gering. Methode: Der Spiraltrokar wurde in vivo und in vitro untersucht und mit anderen Trokatypen verglichen. Ergebnisse: Er wies ein hohes Sicherheitsniveau und gute Handlingeigenschaften auf. Schlussfolgerung: Da es sich um ein einfach zu reinigendes Mehrwegprodukt ohne mechanische Verschleissteile handelt, dass mit den meisten Arbeitshülsen kompatibel ist, besteht auch eine hohe Kosteneffizienz.      

Serum profiles of insulin-like growth factors and their binding proteins in adults with growth hormone receptor deficiency treated with insulin like growth factor I

Six adult patients with growth hormone receptor deficiency (GHRD) (2 men, 4 women) with an identical defect in the growth hormone receptor (GHR) gene, were treated with recombinant human insulin-like growth factor I (IGF-I), 40 μgikg S.C. twice daily, for 7 days. Serum concentrations of IGF peptide and IGF binding protein-3 (IGFBP-3) were measured by specific radioimmunoassays; serum IGFBPs were also measured by Western ligand blotting. The size distribution of both IGF-I and IGF-II was measured in serum following size-exclusion fast-performance liquid chromatography. IGF-I treatment resulted in a normalization of serum IGF-I levels on days 1–7 of treatment and a decrease in serum IGF-II levels. The fall in IGF-II levels and the simultaneous rise in IGF-I levels, however, resulted in an unchanged total serum IGF level. The low IGFBP-3 values did not significantly change during treatment, whereas there was a slight increase in IGFBP-2 levels. Preliminary analysis of size-fractionated sera suggested an increase in IGF-I levels in the 40 and 150 kDa regions at the expense of IGF-II levels. The results suggest that despite the failure of IGF-I treatment to increase IGFBPs significantly, serum IGFBP concentrations were sufficient to maintain normal levels of IGF-I. 0 Laron syndrome, growth hormone receptor deficiency, insulin-like growth factors, insulin-like growth factor binding protein