Tumor induction by ras and myc oncogenes in fetal and neonatal brain: modulating effects of developmental stage and retroviral dose

Introduction into fetal rat brain cells of a replication-defective retroviral vector harboring v-Ha-ras and v-gag-myc rapidly causes the induction of highly malignant undifferentiated neuroectodermal tumors following transplantation into the brains of syngeneic hosts [Wiestler, et al. (1992) Cancer Res. 52: 3760–3767]. In the present study, we have investigated the modulating effect of the developmental stage of neural target cells and of the dose of the retroviral vector used in the grafting experiments. Exposure of fetal cells from embryonic day (E)12 or E14 produced a 100% incidence of malignant neuroectodermal tumors which led to the death of recipient animals after a median latency period of 32 days. A 100-fold reduction of the virus dose from 2.062×10⁶ to 2.062×10⁴ focus-forming units/ml resulted in a lower tumor incidence of 25%. Of six neural grafts exposed to v-Ha-ras and v-myc at E16, only one showed evidence of tumorigenesis (low-grade astrocytoma and hemangioma). All other transplants were morphologically normal for observation periods of 26 weeks, indicating a marked loss of transforming activity of ras and myc in more advanced stages of brain development. In retrovirus-exposed donor cells which caused the development of neural tumors in recipient rats, malignant transformation was also evident during culture in vitro, usually after 9–12 days. Oncogene complementation was also studied in the newborn rat brain. After microinjection of the retroviral vector into the brain at postnatal day (P)0, P1 and P3, 5 out of 20 animals (25%) developed a total of seven brain tumors. Histopathologically, three of these neoplasms were malignant neuroectodermal tumors which, in contrast to those induced in fetal brain transplants showed evidence of focal glial and/or neuronal differentiation. In addition, we observed one oligodendroglioma, two hemangiomas and a malignant hemangioendothelioma. These data indicate that neural precursor cells and endothelia of the rat brain represent the major target cells for the complementary action of ras and myc and that the use of target cells from later developmental stages (E16 and postnatal) leads to the induction of both primitive and more differentiated neoplasms.These studies were supported by the Fonds zur Förderung der wissenschaftlichen Forschung in Österreich (Erwin Schrödinger fellowship, JO501-MED), by the Swiss National Science Foundation and by the Cancer League of the Kanton of Zürich     

Expression of rabbit Ig allotypes in litters of mothers immunized against a paternal allotype.

Sera of successive littermates of mothers producing anti-allotype antibodies (Ab) were analysed for altered a locus or b locus allotype expression. We measured the allotype concentration in sera of 66 individuals (17 litters) of seven mothers producing anti-a1 Ab, and 63 individuals (15 litters) of seven mothers producing anti-b4 Ab, in an enzyme-linked immunosorbent assay (ELISA). We confirmed that the ability to induce allotype suppression in utero increases with the number of antigen boosts applied to the mother, even though the Ab titre in the maternal serum may be decreased. All individuals of a litter expressed the allotype in about equal concentration. This contrasts the results we obtained when newborn rabbits were injected with anti-allotype antiserum. Injection of the same amount of anti-allotype antiserum into nine offspring of two mothers caused allotype suppression in only five individuals, showing no effect in the others. No suppression was observed when IgG-depleted antiserum was injected into newborn rabbits. As expected, maternal antibodies to a paternal allotype do not affect the Mendelian distribution of the progeny phenotypes.     

Molecular determinants of cetuximab efficacy.

PURPOSE: To investigate whether mRNA expression levels of cyclin D1 (CCND1), cyclooxygenase 2 (Cox-2), epidermal growth factor receptor (EGFR), interleukin 8 (IL-8), and vascular endothelial growth factor (VEGF), all members of the EGFR signaling pathway, are associated with clinical outcome in patients with EGFR-expressing metastatic colorectal cancer (CRC) treated with cetuximab. PATIENTS AND METHODS: Thirty-nine patients with metastatic CRC, refractory to both irinotecan and oxaliplatin, were enrolled on IMCL-0144 and treated with single-agent cetuximab. The intratumoral mRNA levels of CCND1, Cox-2, EGFR, IL-8, and VEGF were assessed from paraffin-embedded tissue samples using laser-capture microdissection and quantitative real-time polymerase chain reaction. RESULTS: There were 21 women and 18 men with a median age of 64 years (range, 35 to 83 years). Higher gene expression levels of VEGF were associated with resistance to cetuximab (P = .038; Kruskal-Wallis test). The combination of low gene expression levels of Cox-2, EGFR, and IL-8 was significantly associated with overall survival (13.5 v 2.3 months; P = .028; log-rank test). Both findings were independent of skin toxicity that was itself significantly correlated to survival. Patients with a lower mRNA amount of EGFR had a longer overall survival compared with patients that had a higher mRNA amount (7.3 v 2.2 months; P = .09; log-rank test). Patients with lower expression of Cox-2 had a significantly higher rate of grade 2 to 3 skin reactions under cetuximab treatment. CONCLUSION: This pilot study suggests that gene expression levels of Cox-2, EGFR, IL-8, and VEGF in patients with metastatic CRC may be useful markers of clinical outcome in single-agent cetuximab treatment.     

Primary Hemiarthroplasty for Proximal Humeral Fractures in the Elderly: Long-Term Functional Outcome and Social Implications

Abstract Background: Primary shoulder hemiarthroplasty is an established treatment modality for complex fractures of the proximal humerus. Long-term functional outcome is often disappointing. However, little is known about social implications particularly in the elderly. Methods: A single-institution case series of consecutive geriatric patients (age > 70 years) treated with shoulder hemiarthroplasty for complex fractures of the proximal humerus between 1994 and 1997 was analysed. Postoperative morbidity, long-term function, radiological outcome and social implications were evaluated. Results: Seventy-seven patients fulfilled the study criteria. Median age at the time of operation was 80 years (range 70–93 years). Systemic and local postoperative complications were observed in 8% including 2 patients (3%) with revision surgery. Postoperative mortality was 1%. Forty-eight patients (62%) were available for follow-up (median 49 months, range 25–80 months), 22 (29%) died from causes unrelated to hemiarthroplasty before follow-up and 7 patients (9%) did not attend follow-up examination. Median Constant-Murley score was 41 points (range 17–77 points). Long-term results concerning pain were satisfying. The Oxford shoulder score ranged from 14 to 40 (median 30). Forty-one patients (85%) still lived in their original environment and managed their daily life independently despite poor shoulder function. Four patients (8%) lived in a retirement home and 3 (6%) in a nursery home. Eighty percent of our patients were still able to use public transportation, do the daily shopping and wash their whole body by themselves. Conclusion: Most patients managed their daily life independently despite poor shoulder function.     

Force recruitment during electrical nerve stimulation with multipolar intrafascicular electrodes

Experimentally, during electrical nerve stimulation, the influence is examined of two intrafascicular anodes on the force recruitment with one intrafascicular cathode. It is found that the anodes suppress recruitment and that this effect is more pronounced when the distance of the anodes to the cathode is decreased, or when the anodal currents are increased. The measured recruitment curve patterns can be qualitatively explained by a nerve stimulation model that calculates theoretical recruitment curves for intrafascicular multi-electrode configurations. Discrepancies between the experimental and the theoretical recruitment curves are seen, but these can be understood by taking into account a non-uniform fibre distribution.     

RA-spezifische Autoantikörper gegen ein 68k-Antigen

Zusammenfassung Die Diagnostik vieler rheumatischer Systemerkrankungen wird heute durch den Nachweis von Autoantik?rpern unterstützt und erleichtert. Für die Serodiagnostik der Rheumatoiden Arthritis (RA) stehen nur die doch wenig spezifischen Rheumafaktoren zur Verfügung. Mit dem Ziel, neue krankheitsspezifische Autoantik?rper nachzuweisen, erfolgte eine besondere Proteinaufarbeitung aus Synovialisbiopsien und anderen Geweben. Western Blots der gewonnenen Proteine wurden eingesetzt, um Seren von RA-Patienten und solchen mit anderen rheumatischen Erkrankungen zu untersuchen. Die signifikanteste Immunreaktion von RA-Patienten richtete sich gegen ein 68k-Antigen, welches vermutlich ubiquit?r exprimiert wird, da es nicht nur in Synovialis, sondern in allen weiteren untersuchten Humangeweben und HeLa-Zellen nachgewiesen werden konnte. Der isoelektrische Punkt liegt bei 5,1, das Protein ist O-glykosyliert und im endoplasmatischen Retikulum und/oder Cytoplasma lokalisiert. Antik?rper gegen dieses 68k-Antigen waren bei 110 von 167 RA-Patienten nachzuweisen, was einer Sensitivit?t von 66% entspricht. Ihr Vorkommen war unabh?ngig vom Rheumafaktornachweis, da sie auch bei 7 von 12 seronegativen RA-Patienten zu finden waren, dagegen nur bei einem Patienten aus einer Kontrollgruppe von 98 Patienten mit anderen rheumatologischen Krankheitsbildern, bei einem von 22 HIV-Patienten und überhaupt nicht bei 55 Gesunden. Daraus resultiert eine RA-Spezifit?t für diesen Antik?rper von 99%. Wegen der auff?lligen Krankheitsspezifit?t der anti-68k-Antik?rper liegt es nahe, nach korrespondierenden autoreaktiven T-Zellen zu suchen, um die Rolle dieser neuen Autoreaktivit?t in dem Pathomechanismus der RA zu analysieren. Eingegangen: 20. Mai 1996 Akzeptiert: 13. Februar 1997