Cloning and sequencing of a tpr-met oncogene cDNA isolated from MNNG-transformed human XP fibroblasts

A rearranged tpr-met oncogene was identified in a MNNG-transformed human Xeroderma pigmentosum (XP) cell line (ASKMN). A 2016 bp cDNA was cloned and sequenced, disclosing an ORF with a coding capacity for a 523 aa protein. The sequence of this tpr-met cDNA was very similar to that previously reported in another human MNNG-transformed cell line (MNNG-HOS).     

Hemocompatibility in hemodialysis and erythropoietin therapy

Two studies designed to investigate the effect of recombinant human erythropoietin (rHuEPO) treatment of anemia in chronic dialysis patients on hemocompatibility were conducted. Study 1, whose main aim was to establish whether treatment with rHuEPO enhances coagulation activation during dialysis, included 15 patients before rHuEPO therapy at a mean hematocrit (HCT) of 22.3% and then during therapy at a HCT of 29.3%. The plasma concentrations of the thrombin-antithrombin III complex were not higher during rHuEPO therapy than before it when performing hemodialysis with a Cuprophan membrane. No significant difference was demonstrated either in the values of activated clotting times (Hemochron), thrombocyte or white blood cell counts (Coulter S+II), or in plasma C5a concentrations (ELISA) established during dialysis sessions before and during rHuEPO therapy. In Study 2, which focused primarily on the question of whether or not rHuEPO therapy increases thrombocyte activation during hemodialysis, 8 patients on chronic dialysis were examined both before therapy at a mean HCT value of 22.1% and during rHuEPO therapy at a HCT of 31.5%, invariably during dialysis with either a Cuprophan or polyacrylonitrile (AN69HF) membrane. The plasma concentrations of beta-thromboglobulin (ELISA) did not differ between the examinations made during rHuEPO and before rHuEPO therapy; however, statistically significant differences were found between dialysis sessions involving Cuprophan and AN69HF membranes. No significant difference between examination before and during rHuEPO was demonstrated in activated clotting time nor thrombocyte and white blood cell counts in this study either. The authors conclude that rHuEPO therapy does not enhance coagulation activation during hemodialysis, does not have an effect on thrombocyte activation, and does not influence complement activation and changes in white blood cell counts.     

A genetic model for undifferentiated cell tumor formation: similar tumors formed by different cell lines transformed by the E1A oncogene

The activation of oncogenes and the mutation/deletion of suppressor genes may be involved in tumor heterogeneity. In an attempt to study tumor heterogeneity, we transformed cell lines from epithelial (PAM 212), mesenchymal (NIH-3T3), or melanocytic origin (L-BIOBR) with the wild type E1a oncogene. To make the cell lines tumorigenic, cells were infected with Harvey sarcoma virus carrying the v-H-ras oncogene. The transformed cells were injected into nude mice and the tumors studied by optical and electron microscopy. The tumors formed by v-H-ras-transformed cells consisted of epitheliod melanomas, spindle cells sarcomas and poorly-differentiated carcinomas, depending on the cell of origin. In contrast, the tumors obtained from cells also carrying the E1a oncogene showed a predominant small and undifferentiated cell pattern regardless of the cell of origin. We conclude that the E1a oncogene products induce a negative control of differentiation, independent of the cell type, and that tumors formed by cells carrying the E1a oncogene display an undifferentiated cell pattern.     

Renopancreatic transplant. Urologic complications

OBJECTIVE: Retrospective study of urological complications in our series of reno-pancreatic transplants. MATERIAL AND METHODS: Between February 1983 and May 1994 our group has conducted 93 RPT, 80 of which, mean age 36 +/- 6 years (24-54 years), are studied in this paper: 57 male and 23 female with an average time in dialysis of 20 +/- 15 months (0-84 months) and diabetes evolution of 21 +/- 5 years (11-37 years). RESULTS: Actuarial annual survival of patient, renal graft and pancreatic graft has been 85%, 79% and 74% respectively. Haematuria: 25% incidence, with graft pancreatitis etiology in 16 cases, rejection in 8 and urinary fistula in 6. Urinary infection: 85% incidence, symptomatic in 23 patients (29%) and asymptomatic in the remaining cases. Dysuria, urethritis and urethral stenosis: 14 patients, all male, most with both conditions associated. Reconversion of pancreatic exocrine secretion by intestinal route was performed in 7 patients. Urinary fistula: secondary to surgery in 9 cases and rejection in 4. Etiology of one case remained unknown. In 4 cases it was resolved with conservative treatment, and with surgical correction in 8. One patient required pancreatic transplantectomy and one patient died of AMI. CONCLUSIONS: Urological complication in RPT account for a significant morbidity, urinary fistula being the one with greater repercussion on the patient and pancreatic graft survival.     

Two-dimensional map of Haemophilus influenzae following protein enrichment by heparin chromatography

Two-dimensional gel electrophoresis separates several hundred protein molecules in one single experiment and is efficiently used to study the products expressed by different genomes. Low-copy-number gene products are invisible on a stained two-dimensional map and must be enriched such that sufficient amounts are present for visualization and identification. We investigated the enrichment of proteins of the bacterium Haemophilus influenzae by chromatography on immobilized heparin which has affinity for growth and protein biosynthesis factors. Total soluble proteins of the microorganism were fractionated on Heparin-Actigel which resulted in enrichment of approximately 160 proteins. The eluates, representing about 40% of the applied proteins, were analyzed by two-dimensional gel electrophoresis and the protein spots were characterized by amino acid composition analysis and matrix-assisted laser desorption ionization mass spectrometry. The proteins enriched by chromatography on the heparin gel were not exclusively low-copy-number gene products and they did not exclusively belong to one single class of proteins. The proteins that bound to the heparin gel are indicated in a two-dimensional protein map which includes more than 110 newly identified proteins.