Epstein-Barr virus (EBV) reactivation in allogeneic stem-cell transplantation: relationship between viral load, EBV-specific T-cell reconstitution and rituximab therapy

BACKGROUND: Monitoring of Epstein-Barr virus (EBV) reactivation after allogeneic hematopoietic stem-cell transplantation markedly improved with quantitative real-time polymerase chain reaction amplification of EBV DNA and visualization of EBV-specific CD8+ T cells with peptide-human leukocyte antigen (HLA) class I tetramers. We decided to combine these methods to evaluate posttransplant EBV reactivation and rituximab therapy. METHODS: We followed 56 patients treated with an HLA-genoidentical sibling (n=32), an HLA-matched unrelated donor (MUD, n=19), or an unrelated cord-blood transplant (n=5). EBV DNA was quantified in plasma and in peripheral blood mononuclear cells (PBMC). Patient CD8+ T cells were stained with a panel of eight tetramers. RESULTS: EBV DNA was detected in half of the patients, mainly in the MUD group (17/19). In 19 patients, viral DNA was detected only in the cellular compartment. All patients who controlled reactivation without rituximab and despite a viral load of greater than 500 genome equivalents (gEq)/150,000 PBMC mounted an EBV-specific CD8+ T-cell response in greater than 1.4% of CD3+CD8+ T cells. Plasmatic EBV genome was found in nine patients preceded by a high cellular viral load. Three of these patients controlled the reactivation before or without the introduction of rituximab, and they all developed a significant and increasing EBV-specific T-cell response. Patients with EBV-specific T cells at the onset of reactivation controlled viral reactivation without rituximab. CONCLUSION: This study emphasizes the benefit of an early and close monitoring of EBV reactivation and CD8+-specific immune responses to initiate rituximab only when necessary and before the immune response becomes overwhelmed by the viral burden.     

Etiology of bacterial infections in febrile neutropenic patients: the role of the laboratory in the diagnosis

EPIDEMIOLOGICAL EVOLUTION: Until the mid-eighties, infectious complications (pneumonia, septicemia) observed in neutropenic patients were, in 70% of cases, of bacterial origin with Gram negative bacillae (Escherichia coli, Klebsiella sp, Pseudomonas aeruginosa) isolated 8 times out of 10. Among the Gram positive bacteria, Staphylococcus aureus predominated. The etiological profile of bacterial infections has since evolved with a predominance (60 to 70%) of Gram positive bacteria (coagulase-negative staphylococci, viridans streptococci) and a change in the epidemiology of the Gram positive bacteria notably with a lesser frequency of P. aeruginosa infections. THE GRAM POSITIVE BACTERIA: Coagulase-negative staphylococci are among the first germs responsible for nosocomial bacteremia (central venous catheters) and they are usually multiresistant. Viridans streptococci are a frequent cause of bacteremia; they are generally sensitive to antibiotics active on Gram positive bacteria, but the incidence of resistant strains is increasing. Enterococci are in majority responsible for colonisation in neutropenic patients and less frequently for infections; they raise the problem of resistance to antibiotics, notably to glycopeptides. Other Gram positive bacteria can be responsible for infections in neutropenic patients; it is crucial that they be identified because they require treatment with an appropriate antibiotic. GRAM NEGATIVE BACTERIA: Among the enterobacteria, Escherichia coli is predominantly isolated and raises the problem of the increasing incidence of resistance to fluoroquinolone. Pseudomonas aeruginosa, less frequently responsible today, remains associated with a far greater rate of mortality than that observed with the other microorganisms. Other Gram negative bacteria can be identified; they require an adapted antibiotherapy because they are often naturally multiresistant to antibiotics. THE ROLE OF THE LABORATORY: For the diagnosis of infections in neutropenic patients, the microbiology laboratory has a determinating role. The laboratory ensures the analysis of various biological examinations: blood cultures, methods permitting the diagnosis of an infection on a permanent catheter, copro-cultures (research for common enteropathogens, quantification in the case of digestive decontamination, screening for multiresistant bacteria), cytobacteriological examination of urine, samples of respiratory origin, cytobacteriological examination of cerebro-spinal fluid...).     

Discrimination of osteoporotic patients with quantitative ultrasound using imaging or non-imaging device

OBJECTIVES: Quantitative ultrasound (QUS) has emerged as a new tool in the assessment of fracture risk. The aim of this study was to compare the clinical utility of QUS parameters measured using imaging and non-imaging devices in the discrimination of osteoporotic patients. METHODS: QUS (Broadband Ultrasound Attenuation, BUA dB/MHz, and Speed of Sound, SOS m/s) were measured and then statistical analyses were performed. RESULTS: The 106 women included were 65 +/- 8 years aged. Using DXA, T score was < or = -2.5 at either lumbar spine or hip in 59% of patients, and 25% had osteoporotic fractures. QUS results were different among devices, and these differences were highly dependent on the measured value. There was a similar effect of age and duration of menopause for all parameters. To obtain 90% of sensitivity for the diagnosis of osteoporosis, the thresholds were 50.80 and 71.70 dB/MHz for BUA and 1544.80 and 1551.50 m/s for SOS, using imaging and non-imaging devices, respectively. Belonging to the highest tertile of QUS had a negative predictive value for osteoporosis ranging from 59% to 65%. In the lowest tertile of QUS, the proportion of osteoporotic women was between 73% and 80%. All QUS parameters, except BUA measured with the non-imaging device, were able to discriminate post-menopausal women with fractures after adjustment for age and hip BMD. CONCLUSIONS: Our data suggest that an imaging system improves the utility of BUA measurement, but not SOS, for post-menopausal osteoporosis assessment.     

Influence of the number of motile spermatozoa inseminated and of their morphology on the success of intrauterine insemination

BACKGROUND: Although intrauterine insemination (IUI) is one of the most common assisted reproductive technology methods in the world, the relative influence of various semen characteristics on the likelihood of a successful outcome is controversial. The aim of our study was to assess the results of IUI as a function of both the number of motile spermatozoa inseminated (NMSI) and the percentage of morphologically normal spermatozoa after preparation. METHODS: This was a retrospective study of 889 couples who underwent 2564 IUI cycles of ovarian stimulation with HMG or recombinant FSH in our centre between January 1991 and December 2000. RESULTS: A total of 331 clinical pregnancies were obtained, for a pregnancy rate/cycle of 12.91%. When the NMSI was < 1 x 10(6), the pregnancy rate/cycle was significantly lower (3.13%) than in any of the subgroups with NMSI > or = 2 x 10(6). Sperm morphology, assessed before or after preparation, was not in itself a significant factor that affected the likelihood of IUI success. Nonetheless, when the post-migration rate of normal sperm was < 30%, the pregnancy rate/cycle was 5.43% when NMSI was < 5 x 10(6) and 18.42% when NMSI was > or = 5 x 10(6) (P = 0.008). Pregnancy rates did not differ significantly according to NMSI when the percentage of normal sperm after preparation was > or = 30%, or according to percentage of normal sperm when the NMSI was > or = 5 x 10(6). CONCLUSIONS: Our results show that a minimum of 5 x 10(6) motile spermatozoa should be inseminated when the normal morphology of the sperm after preparation is < 30%; the quantity compensates at least in part for the defective quality. If this threshold of NMSI cannot be obtained, IVF should be recommended.     

Quantitative body composition analysis in awake mice and rats by magnetic resonance relaxometry

OBJECTIVE: Magnetic resonance (MR) relaxometry has recently been introduced for noninvasive body composition analysis in awake mice. The purpose of the present study was to extend the method to rats and to introduce calibration procedures that render MR relaxometry fully quantitative. RESEARCH METHODS AND PROCEDURES: Proton T(2) MR relaxometry at 4.7 Tesla was used for body composition analyses in 700 awake mice and 400 rats of different strains and conditions. Relaxograms calculated from the signal decays observed with multi-spin-echo acquisition provided well-separated contributions of tissue water and fat. Analysis of fat composition was carried out in vivo using (13)C-MR spectroscopy. Evolution of body composition in rats was assessed during drug treatment. RESULTS: MR relaxometry for noninvasive body composition analysis in laboratory rodents was implemented on a standard MR scanner, and a throughput of >30 animals per hour was achieved. Excellent linearity and reproducibility with coefficients of variance as low as 2.5% and 1.7% were obtained in mice and rats, respectively. The lean mass-to-water ratio (mice, 1.35 +/- 0.03; rats, 1.39 +/- 0.04) and the proton density of fat (mice, 8.1 +/- 0.2; rats, 8.9 +/- 0.2 g/mol) were determined from cross-sectional data. Fat composition analysis by (13)C-MR spectroscopy corroborated these findings and yielded information on the average acyl chain length (16.3 +/- 1.6) and contributions of saturated (27 +/- 3%), monounsaturated (22 +/- 2%), and polyunsaturated (51 +/- 3%) fatty acids. Longitudinal assessments in rats treated with sibutramine and dexfenfluramine showed dose-related changes in body composition. DISCUSSION: T(2) MR relaxometry backed by solid calibration provides a powerful means for rapid quantitative body composition analysis in awake mice and rats that is suitable for serial investigations in pharmaceutical research.     

Drugs under temporary use authorisation: 50 years of French drug development

Drug under temporary use authorisation (authorisation temporaire d' utilisation - ATU) was created in 1994 in France. Before this date, various procedures controlled patients' use of imported drug and drug in process. The AIDS pandemy development in 80's and the need to control news drugs patients' use contributed to the ATU statement setting up. This regulatory statement allows companies to sell drugs without market drug authorisation (autorisation de mise sur le marché - AMM), the letter being done in adequation with good pharmaceutical quality and a satisfactory level of benefices/ risks ratio. It's a recent French drug evaluation evolution.     

Absence or low expression of fas-associated protein with death domain in acute myeloid leukemia cells predicts resistance to chemotherapy and poor outcome

In acute myeloid leukemia (AML), coexpression of death receptors and ligands of the tumor necrosis factor (TNF) receptor/TNF-alpha superfamily on leukemic cells after chemotherapy is not always accompanied by apoptosis, suggesting that the apoptotic death receptor signaling pathway is disrupted. Because Fas-associated protein with death domain (FADD) is the main adaptor for transmitting the Fas, TNF-related apoptosis-inducing ligand receptors, and TNF receptor 1 death signal, expression of FADD was analyzed by Western blot and immunocytochemistry in leukemic cells of 70 de novo AML patients treated with the European Organization of Research and Treatment of Cancer AML-10 randomized trial before initiation of induction chemotherapy. Thirty seven percent of patients (17 of 46) with FADD negative/low (FADD(-/low)) leukemic cells had a primary refractory disease compared with 12% of FADD(+) patients (3 of 24; P = 0.05). FADD(-/low) expression was significantly associated with a worse event-free survival [EFS (P = 0.04)] and overall survival (P = 0.04). In multivariate analysis, FADD(-/low) protein expression was independently associated with a poor EFS and overall survival (P = 0.002 and P = 0.026, respectively). Importantly, FADD(-/low) protein expression predicted poor EFS even in patients with standard- or good-risk AML (P = 0.009). Thus, we identified low or absent expression of the FADD protein in leukemic cells at diagnosis as a poor independent prognostic factor that can predict worse clinical outcome even for patients with standard- or good-risk AML.     

Clinical and immunohistochemical analysis of patients with unknown primary tumour. A search for prognostic factors in UPT

BACKGROUND: The unknown primary tumour (UPT) is an intriguing clinical finding in approximately 5% of all newly diagnosed patients with cancer. To evaluate a correlation between the specific immunohistochemical alterations in UPT cells and the unique clinical features of UPT patients, to define the natural history of UPT and to verify prognostic factors, we undertook a detailed clinical and immunohistochemical analysis of patients with the diagnosis of adenocarcinoma of UPT. RESULTS: Patients with UPT present with a short history and have a poor prognosis. Univariate analysis was performed with clinical, biological and immunohistochemical variables. Patients with a higher age (>60 years), a poor performance score (2-3), liver metastases or more than two organ sites involved, or patients with elevated LDH-levels, were found to have worse prognosis. We confirm that the prognostic model published by Culine is a valuable model for the prediction of prognosis in patients with UPT. Immunohistochemical detection of proliferation (MIB-1), p53, vascular endothelial growth factor-A, CD34, CD44v6 and Her2neu indicated that these factors were of no prognostic value. CONCLUSION: In conclusion, patients with UPT have a very poor median prognosis of 12 weeks. Prognostically favourable factors are young age, good performance status, no liver metastases and normal LDH level. We found no relationship with immunohistochemical factors.     

Gastric metastasis from ovarian carcinoma revealed by a gastro-splenic perforation

Metastatic disease involving the stomach is unusual. We report the case of a gastric metastasis from ovarian cancer revealed by gastro-splenic perforation. The gastric metastasis was diagnosed 17 years after the diagnosis of primary cancer.