Development of a biochip-based assay integrated in a global strategy for identification of fusion transcripts in acute myeloid leukemia: a work flow for acute myeloid leukemia diagnosis.

Three major types of rearrangements are involved in acute myeloid leukemias (AML): t(8;21)(q22;q22), inv(16)(p13q22), and 11q23/MLL abnormalities. Their precise identification becomes essential for diagnosis, prognosis, and therapeutic choices. Resulting fusion transcripts (FT) are also powerful markers for monitoring the efficacy of treatment, the minimal residual disease (MRD) and could become therapeutic targets. Today, the challenge is to propose an individual follow-up for each patient even for those with a rare fusion event. In this study, we propose a biochip-based assay integrated in a global strategy for identification of rare FT in AML, after fluorescence in situ hybridization detection, as described by the World Health Organization classification. Using cell lines, we developed and validated a biochip-based assay called the AMLFusionChip that identifies every FT of AML1-ETO, CBFbeta-MYH11 as well as MLL-AF9, MLL-ENL, MLL-AF6, and MLL-AF10. The original design of our AMLFusionChip.v01 enables the identification of these FT wherever the breakpoint on the partner gene may be. In case of biochip negative result, our 3'RACE amplification strategy enables to clone and then sequence the new translocation partner. This AMLFusionChip strategy fits into the concept of personalized medicine for the largest number of patients.

[Molecular typing of breast cancer: transcriptomics and DNA microarrays]. / Typage moléculaire du cancer du sein: transcriptome et puces á ADN.

Breast cancer is the most frequent and deadly cancer of women. Its great heterogeneity makes prognosis and response to current treatments highly variable and difficult to predict. Mammary oncogenesis remains poorly understood. These issues should benefit from recent development of techniques capable of large-scale molecular analyses. The use of cDNA array techniques allows for the simultaneous analysis of the mRNA expression levels of thousands of genes in mammary tumor cell lines and breast tumors. Expression profiles will help classify tumors and provide new prognostic tools and potential therapeutic targets. They will also boost our knowledge of the molecular events responsible for the development and progression of this cancer.

Gene expression profiling of primary breast carcinomas using arrays of candidate genes.

Breast cancer is characterized by an important histoclinical heterogeneity that currently hampers the selection of the most appropriate treatment for each case. This problem could be solved by the identification of new parameters that better predict the natural history of the disease and its sensitivity to treatment. A large-scale molecular characterization of breast cancer could help in this context. Using cDNA arrays, we studied the quantitative mRNA expression levels of 176 candidate genes in 34 primary breast carcinomas along three directions: comparison of tumor samples, correlations of molecular data with conventional histoclinical prognostic features and gene correlations. The study evidenced extensive heterogeneity of breast tumors at the transcriptional level. A hierarchical clustering algorithm identified two molecularly distinct subgroups of tumors characterized by a different clinical outcome after chemotherapy. This outcome could not have been predicted by the commonly used histoclinical parameters. No correlation was found with the age of patients, tumor size, histological type and grade. However, expression of genes was differential in tumors with lymph node metastasis and according to the estrogen receptor status; ERBB2 expression was strongly correlated with the lymph node status (P < 0.0001) and that of GATA3 with the presence of estrogen receptors (P < 0.001). Thus, our results identified new ways to group tumors according to outcome and new potential targets of carcinogenesis. They show that the systematic use of cDNA array testing holds great promise to improve the classification of breast cancer in terms of prognosis and chemosensitivity and to provide new potential therapeutic targets.

Distinct regulation of T-cell death by CD28 depending on both its aggregation and T-cell receptor triggering: a role for Fas-FasL.

CD28 is a major coreceptor that regulates cell proliferation, anergy, and viability of T cells. The negative selection by T-cell receptor (TCR)-induced cell death of immature thymocytes as well as of activated human antigen-specific T-cell clone, requires a costimulatory signal that can be provided by CD28. Conversely, CD28-mediated signals increase expression of Bcl-XL, a survival gene, and promote survival of naive T cells cultured in the absence of antigen or costimulation. Because CD28 appears to both protect from, or induce T-cell death, one important question is to define the activation and cellular parameters that dictate the differential role of CD28 in T-cell apoptosis. Here, we compared different CD28 ligands for their ability to regulate TCR-induced cell death of a murine T-cell hybridoma. In these cells, TCR triggering induced expression of Fas and FasL, and cell death was prevented by anti-Fas blocking monoclonal antibody (MoAb). When provided as a costimulus, both CD28 MoAb and the B7.1 and B7.2 counter receptors downregulated, yet did not completely abolish T-cell receptor-induced apoptosis. This CD28 cosignal resulted in both upregulation of Bcl-XL and prevention of FasL expression. In marked contrast, when given as a single signal, CD28 MoAb or B7.1 and B7.2 induced FasL expression and resulted in T-cell death by apoptosis, which was dependent on the level of CD28 ligation. Furthermore, triggering of CD28 upregulated FasL and induced a marked T-cell death of previously activated normal peripheral T cells. Our results identify Fas and FasL as crucial targets of CD28 in T-cell death regulation and show that within the same cell population, depending on its engagement as a single signal or as a costimulus together with the TCR, CD28 can either induce a dose-dependent death signal or protect from cell death, respectively. These data provide important insights into the role of CD28 in T-cell homeostasis and its possible implication in neoplastic disorders.

Physical and functional interaction of Nef with Lck. HIV-1 Nef-induced T-cell signaling defects.

The nef gene is unique to the primate lentiviruses and encodes a cytoplasmic membrane-associated protein that affects T-cell signaling and is essential for both maintenance of a high virus load in vivo and for disease progression. Here we investigated the perturbation of cell signaling by Nef in T-cells and found that Nef interacts with the T-cell restricted Lek tyrosine kinase both in vitro and in vivo. The molecular basis for this interaction was analyzed. We show that cell-derived Nef is precipitated in a synergistic manner by the recombinant Src homology 2 (SH2) and SH3 domains from Lck. A functional proline-rich motif and the tyrosine phosphorylation of Nef were evidenced as likely participants in this interaction. The precipitation of Nef by the Lck recombinant proteins was specific, since neither Fyn, Csk, p85 phosphatidylinositol 3-kinase nor phospholipase Cgamma SH2 domains coprecipitated Nef from T-cells. Finally, depressed Lck kinase activity resulted from the presence of Nef, both in vitro and in intact cells, and nef expression resulted in impairment of both proximal and distal Lck-mediated signaling events. These results provide a molecular basis for the Nef-induced T-cell signaling defect and its role in AIDS pathogenesis.

Indices of adequate dialysis in patients hemodialyzed with AN 69 membrane.

The objective of this study was to evaluate the incidence of morbidity (at least one hospitalization) during the first twelve months of hemodialysis (thrice weekly for 4 hours) in 54 (27 males and 27 females) sex and age matched patients, of whom 32 were treated with AN 69 (M/F = 13/19, 62 +/- 14 years) and 22 with Cuprophan (M/F = 14/8, 61 +/- 14 years). Patients were classified according to the value of TAC urea during the period under study: constantly superior or equal to 20 mmol/liter in Group A (high TAC urea) or inferior to 20 mmol/liter in Group B (low TAC urea). Dialysis quantification (Kt/V) and estimation of the patient's protein catabolic rate (PCR) were based on measurement of the midweek pre- and post-dialysis blood urea nitrogen. In the patients of Group B, incidence of morbidity was significantly increased when age was over 50 years and when AN 69 membrane was used (P < 0.02). Furthermore, in Group A, the risk of hospitalization was significantly higher in patients treated by Cuprophan than in those treated by AN 69 (P < 0.02). The survival rate was also studied. Better survival (70%) at four years was observed in patients with high TAC urea who were treated by AN 69. The difference was highly significant with the survival rate (22%) in patients with high TAC urea who were treated by Cuprophan (P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

[Genetic and environmental factors implicated in recurrent calcium lithiasis]. / Facteurs génétiques et environnementaux impliqués dans la lithiase calcique récidivante.

In this prospective study, we compared the frequency of some genetic and environmental factors possibly implicated in the occurrence of calcium stone disease. A group of 439 patients (258 males and 181 females) with one episode of calcium stone was compared to a group of 191 patients (131 males and 60 females) with recurrent calcium stone disease. Population with stones was also compared to control population (n = 78, 40 males and 38 females) matched to age. Major results were as follow: 1) Family history of urinary calculi was more frequent in patients than in controls (28.4% vs 9%, p < 0.01). No difference was observed between patients with one episode and those with recurrent episodes (27% vs 31%, ns). 2) The recurrence was earlier in female than in male, so that in female with family history of urinary calculi (p < 0.05). 3) Mean plasma levels of 1-25OH2D3 was significantly higher in patients with family history than in controls (60% vs 38%, p < 0.01) 5) Restricted calcium diet (< 400 mg per day) was more often observed in patients than in controls (31% vs 14%, p < 0.05) and the most significant difference was found in patients with recurrent calcium stones.

[Short and long term complications of lithotripsy on renal function]. / Séquelles à court et moyen termes de la lithotripsie sur la fonction rénale.

Today, most stones can be removed by minimally invasive means. Extracorporeal Shock Wave Lithotripsy (ESWL) is the preferred form of treatment for symptomatic upper ureteral and renal calculi less than 2 cm a diameter. The short and long term complications of ESWL are underestimated. Thus, ESWL may cause renal trauma and such trauma may induce later hypertension. In this retrospective study, we reviewed the frequency of deleterious effects of ESWL in 45 patients who had undergone ESWL from January 1988 to September 1989. Short-term complications were macroscopic hematuria (15%), lumbar pain (11%) and peri- or intrarenal hematomas (4.4%). Two years later CT scan was performed in 20 patients. It was normal in 7 (35%). In others, it shown a recurrence of stone in 8 (40%) and a focal scarring in 5 (25%). Only 1 out of 43 patients had developed hypertension.

[Isolated systolic hypertension in uremic hemodialyzed patients]. / Hypertension artérielle systolique isolée chez l'urémique hémodialysé.

Prevalence of Isolated Systolic Hypertension (ISH) defined as systolic blood pressure greater than 160 mmHg and diastolic blood pressure less than 90 mmHg was studied in a population of 148 patients treated by hemodialysis whose 80 had undergone ambulatory blood pressure (ABP) recording during the interdialytic period. All patients were treated 3 times 4 hours a week. ABP was recorded for 48 hours between two sessions of hemodialysis using a Delmar Avionic Presurometer (PIV). Prevalence of ISH was 12.5%, while that of systolic-diastolic hypertension (SDH) was 15%. Average age at the time of the study was respectively 59 +/- 13 yrs ISH and 49 +/- 11 yrs SDH (p less than 0.01) while that of patients with normal blood pressure (N) was 57 +/- 10 yrs. Mean duration of HD treatment was no different between groups: 5.3 +/- 3.5 yrs ISH, 5.0 +/- 4.2 yrs SDH and 5.0 +/- 4.3 yrs N. Causes of end-stage renal disease were similar in each group. All patients with ISH and SDH and 42% of N pts were receiving antihypertensive treatment at the time of ABP recording. Finally, level of anemia and percentage of patients treated by EPO were similar in each group.(ABSTRACT TRUNCATED AT 250 WORDS)