An immunohistochemical study of the simultaneous expression of bcl-2 and p53 oncoproteins in epithelial tumors of the colon and rectum.

OBJECTIVE: Bcl-2 and p53 genes may participate in a common pathway for regulation of apoptosis. The aims of this study were (1) to study the immunohistochemical expression of the bcl-2 oncoprotein in colorectal tumors, (2) to correlate it with that of p53 protein overexpression, and (3) to compare it with histopathologic prognostic factors, such as TNM classification and grade. DESIGN: Prospective study of expression of bcl-2 and p53 oncogenes in colorectal tumors. We examined 6 colorectal hyperplastic polyps, 33 adenomas, and 61 carcinomas. SETTING: Regional academic medical center. METHODS: An immunohistochemical study with bcl-2 and p53 antibodies was performed on frozen sections of colorectal tumors. The levels of bcl-2 and p53 expression were evaluated using a semiquantitative grading system. Two-color immunohistochemistry was performed to examine the intracellular colocalization of bcl-2 and p53 in all tumors with a strong positivity for both antigens. RESULTS: Bcl-2 was expressed in 28 (85%) of the 33 adenomas, whereas p53 was expressed in only one adenoma, which had areas of in situ carcinoma. Bcl-2 and p53 were each expressed in 43 (70.4%) of the 61 carcinomas. Thirty-one (50%) of the colorectal carcinomas coexpressed the two oncoproteins. There was no correlation between the number of cells expressing bcl-2 and the number expressing p53 in a given carcinoma. No correlation was observed between the expression of bcl-2 or p53 and the established prognostic factor. CONCLUSION: Abnormal bcl-2 oncoprotein expression appears earlier than p53 accumulation in colorectal carcinogenesis. This study suggests that there is more than one sequence and mechanism of bcl-2 and p53 gene deregulation in colorectal carcinomas.

Effects of pentoxifylline on circulating cytokine concentrations and hemodynamics in patients with septic shock: results from a double-blind, randomized, placebo-controlled study.

OBJECTIVE: To determine whether a continuous intravenous infusion of pentoxifylline, a methylxanthine derivative, alters the serum cytokine concentrations and/or hemodynamic measurements in patients with septic shock. DESIGN: A prospective, randomized, double-blind, placebo-controlled study. SETTING: Medical intensive care unit in a university hospital. PATIENTS: Sixteen patients with septic shock. INTERVENTIONS: Patients were randomly assigned to receive either pentoxifylline (1 mg/kg) followed by an infusion of 1.5 mg/kg/hr for 24 hrs (n = 8), or placebo (n = 8). MEASUREMENTS AND MAIN RESULTS: Tumor necrosis factor (TNF) and interleukin (IL)-6 concentrations were measured by radioimmunoassays; IL-8 concentrations by an enzyme-linked immunosorbent assay (ELISA) and pentoxifylline concentrations by high-performance liquid chromatography at 0, 3, 6, 12, 18, 24 and 48 hrs after study entry. Pulmonary artery catheter-derived hemodynamics were measured at 0, 0.75, 3, 6, 12, 18, and 24 hrs. In pentoxifylline-treated patients, at 24 hrs, serum concentrations of TNF were significantly lower compared with controls (12 +/- 2 vs. 42 +/- 12 pg/mL, respectively, p = .04). Serum concentrations of IL-6 and IL-8 did not differ between the two treatment groups. There were also no significant differences in any hemodynamic and oxygenation measurements comparing the two treatment groups. Pentoxifylline concentrations were 1,544 +/- 241 ng/mL after the initial dose, and 5,776 +/- 1,781 ng/mL at the end of the 24-hr infusion. Five patients in the pentoxifylline group and four patients in the placebo group died. CONCLUSIONS: Pentoxifylline is able to decrease serum TNF but not IL-6 or IL-8 serum concentrations during septic shock. Pentoxifylline was well tolerated by all eight patients with no adverse effect. Further studies are needed to determine if pentoxifylline's ability to lower circulating TNF concentration without altering hemodynamics will improve outcome in septic shock.

[Follow-up of allogenic bone marrow graft with DNA polymorphisms. Use of two minisatellite MS31 and MS43 probes. Comparison with data of chromosomal polymorphisms]. / Suivi de greffe allogénique de moelle osseuse par les polymorphismes de l'ADN. Utilisation de deux sondes minisatellites MS 31 et MS 43. Comparaison avec les données des polymorphismes chromosomiques.

OBJECTIVES: Allogenic bone marrow transplantation is widely used to treat many diseases of the haemopoietic system as well as metabolic disorders. Follow-up is essential to assess acceptance, rejection or post-graft relapse. This study was undertaken to evaluate the usefulness of the minisatellite probes MS31 and MS43 used as a routine follow-up test after bone marrow transplantation. METHODS: Twenty receivers of allogenic bone marrow transplants were followed-up. Two monoclonal minisatellite probes, MS31 and MS43, were used for comparison with the classical polymorphism methods. RESULTS: Fourteen cases of total chimeras, 3 cases of rejections and 3 cases of mixed chimeras were observed with the molecular probe techniques. In 19 of the 20 cases, this technique gave results compatible with classical polymorphism results. CONCLUSIONS: The minisatellite probes MS31 and MS43 were found to be sensitive, effective tests for bone marrow transplants which can be used in routine follow-up.

[Immunoglobulins E and human immunodeficiency virus infection (HIV)]. / Immunoglobines E et infection par le virus de l'immunodéficience humaine (VIH).

Except for Ig E, serum immunoglobulin abnormalities in persons with human immunodeficiency virus (HIV) infection have been well described. Serum IgE levels have been shown to rise with progressive disease. The authors evaluated IgE in 148 HIV-seropositive individuals with or without acquired immunodeficiency syndrome (AIDS). Mean serum IgE levels were compared between groups based on absolute CD4 lymphocyte counts or clinical status (CDC) and with a seronegative control group. Higher serum IgE levels were observed in seropositive-patients. A rise in IgE serum is common in patients with HIV infection; it could be link with an earlier dysregulation in the IgE synthesis. No correlation was found between IgE level and CD4 counts.

Natural serum TNF antagonists in end-stage renal failure and following renal transplantation.

TNF is clearly involved in allograft rejection but measurement of serum cytokine levels do not reflect reliably the rejection crisis. TNF induces release of soluble receptor parts that are more stable, are catabolized by kidneys and have inhibitory activity. Thus it is crucial to analyse their kinetics during renal function recovery after transplantation to forecast their potential clinical use in rejection monitoring or treatment. A sequential study was performed in 61 patients undergoing kidney graft, compared to 60 graft patients with long follow-up and 15 healthy controls. Soluble TNF and Il2 receptors were measured by ELISA and TNF was measured by RIA. The sTNF-Rs were markedly increased in renal failure, much more than another soluble cytokine receptor, the sIl2-R. Levels progressively decreased with the recovery of renal function and became directly correlated to the renal function. Normal levels were only reached after some weeks. No significant changes were observed during graft rejection at that stage of renal function but p75 were higher after antilympocyte antiserum and tubular necrosis. High sTNF-R did not seem to minimize rejection risk or gravity. In long-standing recipients, sTNF-R rose in some patients, particularly with glomerulonephritis, and may help in monitoring chronic rejection activity. sTNF-R but not sIl2-R markedly accumulate in ESRF and mask any changes that could be helpful in the monitoring of early graft events. However, increased levels above renal impairment (sTNF-R/SCr) may be clinically relevant in late rejection or glomerulonephritides.

Detection of quantitative polymerase chain reaction products by hybridization on magnetic support with 125I-radiolabeled probes: quantification of c-myc copy numbers.

We present a technique for determining c-myc copy numbers that can be used as a prognosis index for some cancers. The method is based on the use of both competitive polymerase chain reaction and hybridization of amplified products. Coamplification was performed directly on cells with a synthetic oligonucleotide used as internal standard. It recognized the same primer set as the target. Coamplified products were captured on streptavidin magnetic beads as solid support using a 5' biotinylated primer. DNA immobilized on this support was denatured with alkali. Each coamplified product (target and reference gene) was further hybridized to two distinct specific oligonucleotide probes. Gene amplification levels were determined using a standard curve obtained by serial dilutions of peripheral blood lymphocytes run along with the experimental samples. This approach provides a rapid (less than 2 days) and reproducible method for evaluating c-myc gene copy number and may be used to quantify any gene. Moreover, its format allows for automation.

High levels of tumor necrosis factor-alpha and interleukin-6 in the ascitic fluid of cirrhotic patients with spontaneous bacterial peritonitis.

We prospectively measured the concentrations of immunoreactive tumor necrosis factor-alpha (TNF-alpha), interleukin-1 (IL-1), and interleukin-6 (IL-6) in the serum and ascitic fluid of 14 alcoholic cirrhotic patients with spontaneous bacterial peritonitis (SBP) and 16 alcoholic cirrhotic patients with sterile ascitic fluid. TNF-alpha levels in ascitic fluid were significantly higher for the patients with SBP: 399.57 +/- 129.23 pg/mL vs. 35.76 +/- 5.57 pg/mL (P < .001). IL-6 levels in ascitic fluid were also significantly higher for the patients with SBP: 170,713 +/- 57,109 ng/mL vs. 5,414 +/- 973 ng/mL (P < .001). By contrast, serum levels of TNF-alpha and IL-6 were just slightly more elevated than normal values. The concentration of IL-1 in the ascitic fluid of all patients was elevated, but there was no difference between patients with SBP or sterile ascites in this respect. In the patients with SBP, levels of TNF-alpha and IL-6 in ascitic fluid decreased during the first 48 hours of antibiotic treatment. Our results suggest that measurements of TNF-alpha and IL-6 in ascitic fluid may become useful markers both for the diagnosis of SBP and for monitoring the treatment of cirrhotic patients.

[Contribution of chemiluminescence for determination of low value TSH in patients with dysthyroidism during negative TRH test]. / Apport de la chimiluminescence au dosage de valeurs basses de TSH chez des patients souffrant de dysthyroïdie lors de test à la TRH non répondant.

TSH values performed by immunoradiometric method were compared with chemiluminescent method in 28 hyperthyroid patients with negative TRH-test. This last method gives a better sensitivity (0.020 mUI/L vs. 0.050 mUI/L) and reproducibility (5% V.C. vs. 40%). The measure of undetectable TSH by immunoradiometric assay becomes possible with chemiluminescence. Due to this increase of sensitivity, the TRH-test needs to be reconsidered and allows to discriminate various degrees of TSH suppression.

Increased interleukin-2 receptor affinity in normal HLA A1 B8 DR3 subjects.

Twenty-two subjects (11 HLA A1 B8 DR3, 11 non-A1 B8 DR3) were tested for the capacity of their lymphocytes to express Tac molecules and interleukin-2 (IL-2) receptors (quantified using radiolabelled IL-2) after mitogen stimulation. Ten of these subjects (five A1 B8 DR3 and five non-A1 B8 DR3) were also tested for the ability of their lymphocytes to proliferate under IL-2 stimulation. A1 B8 DR3 subjects express a normal number of high-affinity IL-2 receptor sites, but the affinity of these receptors sites is significantly increased. Unexpectedly, A1 B8 DR3 lymphoblasts show a lower response to IL-2 than non-A1 B8 DR3 for high doses of recombinant IL-2.

[Pre-treatment serum levels of SCC antigen in epidermoid carcinoma of the upper respiratory and digestive tract]. / Antigénémie SCC pré-thérapeutique et carcinomes épidermoïdes des voies aéro-digestives supérieures.

Results from pre-treatment SCC antigen assay were assessed in 106 patients with epidermoid carcinoma of the upper aerodigestive ways. The control population consisted of healthy blood donors (N = 61). Blood antigen levels ranged 1.6 ng/ml to 21.5 ng/ml in the patient population. 35 patients in 106 (i.e., 33% of cases) had levels considered pathological (greater than 2.00 ng/ml). Antigen levels were higher with increasing tumor size and when adenopathy was more marked (N+, N+R+). However, no correlation could be found with the T (stage)--N (histological) classification of tumors or with the site of lesion. Tumor immunologic response was obviously not uniform. Although the SCC antigen presents no diagnostic value, it appears to bear some prognostic significance, regardless of the tumoral stage. Antigen levels below 2.00 ng/ml correlate with (p less than 0.001) good immediate therapeutical results. On the other hand, serum levels greater than 2.00 ng/ml correlate either with non-sterilization, or with locoregional recurrence and/or rapid development of metastases. Other studies are required to confirm these data, and to demonstrate the value of long-term SCC antigen monitoring in these patients.

Molecular diagnosis of chronic myeloid leukemia using a 3'bcr probe.

Chronic myeloid leukemia (CML) is characterized by the Philadelphia chromosome which results from a reciprocal (9; 22) translocation, with the protooncogene c-abl moving from chromosome 9 to 22 and juxtaposed to the proximal bcr. Breakpoints on chromosome 22 are localized within 5.8 kb of the breakpoint cluster region (bcr). We have assessed the feasibility of using a 3'bcr probe for molecular diagnosis of CML. Thirty patients with Ph chromosome negative or positive CML were studied by Southern blot. A bcr rearrangement was seen to be present in all but one patient with Ph+CML. A case of Ph negative CML showed a bcr rearrangement. We conclude that this technique is efficient for molecular diagnosis of CML.

[CA 19-9 in early cancers of the bronchi. Comparison with the carcinoembryogenic antigen]. / Le CA 19-9 dans les cancers primitifs des bronches. Comparison avec l'antigène carcino-embryonnaire.

CA 19-9 (monosialoganglioside) isolated from adenocarcinomas of the gastrointestinal tract can be measured in biological fluids using a monoclonal antibody assay. A radioimmunometric assay kit produced by ORIS Industrie has been used to measure the serum level of CA 19-9 in lung cancer and the results have been compared to that of carcinoembryonic antigen. The combination of the 2 markers increase by 10% the number of subjects with a raised marker. There is no significant relationship between the levels of CA 19-9, the type of tumour or the tumour stage. The recognition of the sialic acid as an epitope by the monoclonal antibody appears to be responsible for the false positive results in non-malignant lung disease.

[IgE antibodies in human Fasciola hepatica distomiasis]. / Etude des anticorps IgE dans la distomatose humaine à Fasciola hepatica.

In patients infected by Fasciola hepatica, total IgE and specific IgE antibodies have been determined by radioimmunoassays, and IgG, IgA, IgM levels by radial immunodiffusion test (Mancini, 1965). Moreover, total and specific IgE levels have been related to parasite egg burden, age, clinical features and eosinophilia. Elevated total IgE and specific IgE antibodies levels have been found respectively in 76% and 48% of the patients whereas there was no significant variations in other immunoglobulins levels. However, though the amount of total and specific IgE was lower than in other helminthic diseases, it appears to be a significant data of the immune response to parasites as it has been reported and discussed previously. It has been shown a significant relationship between total and specific IgE levels, the number of lines by immunoelectrophoresis, and the results of the indirect haemagglutination and indirect fluorescent antibody techniques; each method appeared to be in equal value to perform the early diagnosis of human Fasciola hepatica. In addition, specific IgE antibodies levels were correlated with eosinophilia specially when it exceeds 15%. This results demonstrate the availability of their measurement in the diagnosis of fascioliasis versus other diseases with marked eosinophilia.