Optimisation of retrospective diagnosis of cytomegalovirus congenital infection from dried blood spots

Out of the 90% of cytomegalovirus (CMV) congenitally infected children that are asymptomatic at birth, 5 to 15% will later develop complications, mainly neurodevelopmental defects and/or deafness. Unfortunately, after the first 2 weeks of life, usual diagnostic techniques for CMV detection (viral culture and serology) are useless to differentiate congenital infection from post-natal acquired infection, whereas detection of viral DNA from dried blood spots (DBS; Guthrie cards), systematically collected from all newborns in the first days of life, has been described for late diagnosis of CMV congenital infection. The aim of our study was to choose and optimise a viral DNA extraction method from DBS and to study if CMV DNA detection is reliable when DBS are stored for 1 year at room temperature or 2 months at 37 degrees C. 10 reference cards (blood collected from CMV seronegative newborns (IgG/IgM negative) were "infected" with serial dilutions of virus and spotted on Guthrie cards) were tested. 3 extraction methods were evaluated, products of PCR were analyzes by agarose gel electrophoresis and quantification of CMV from DBS was also performed. Analysis of the results obtained from reference cards showed higher sensitivity of phenol/chloroform extraction following treatment with proteinase K, compared to heat extraction in cell culture medium or extraction with a commercial kit. We did not observe quantitative loss of viral DNA after 1 year storage at room temperature. CMV DNA detection from Guthrie cards could become a very useful tool for retrospective diagnosis of congenital CMV infection when sequelae are diagnosed in the first years of life. We are pursuing this study with DBS from congenitally infected children.     

Diagnostic accuracy of serum asialo-alpha1-acid glycoprotein concentration for the differential diagnosis of liver cirrhosis and hepatocellular carcinoma

BACKGROUND: Serum asialoglycoproteins concentration are increased in patients with hepatic disease. We developed an antibody-lectin sandwich assay that is sensitive and specific to measure asialo-alpha(1)-acid glycoprotein (AsAGP) concentration in human serum and evaluated it as a biochemical marker for hepatic disease. METHODS: Serum AsAGP concentration was measured by antibody-lectin sandwich assay with 610 serum specimens of patients with hepatic disease. Serum from 41 healthy donors and 155 patients with non-hepatic disease served as negative controls. The AsAGP values were analyzed by receiver operator characteristics (ROC) curve analysis. The diagnostic accuracy of AsAGP value was compared with those of the conventional biochemical markers in the liver function test. RESULTS: Serum AsAGP concentration in 83% of patients with liver cirrhosis (LC) and 89% of patients with hepatocellular carcinoma (HCC) was increased over the cutoff value (1.33 microg/ml), indicating that an increase of serum AsAGP concentration is restricted to LC or HCC cases. The area under curve (AUC) in the ROC curve was 0.919 for LC and 0.946 for HCC. CONCLUSIONS: Serum AsAGP concentration exhibited good diagnostic accuracy as a biochemical marker for LC and HCC. The addition of AsAGP to conventional liver function tests may significantly improve the diagnosis and prognosis.     

泽泻汤加味治疗梅尼埃病38例临床观察

目的：观察泽泻汤加味治疗梅尼埃病的临床疗效。方法：将73例梅尼埃病患者随机分为观察组38例和对照组35例,对照组所有患者根据病情予以倍他司汀注射液静脉滴注,并配以减轻水肿、镇静、止呕等对症治疗;观察组使用泽泻汤加味,并随症加减,1剂/d,早晚2次分服。2组均以发作期的治疗为主,5天为1个疗程,共2个疗程。结果：第1个疗程后总有效率观察组为68.42%,对照组为60.00%,2组相比差异有统计学意义（P〈0.05）。第2个疗程后总有效率观察组为89.47%,对照组为77.14%,2组相比差异有统计学意义（P〈0.05）。不良反应发生率观察组为28.57%,对照组为10.53%。2组相比差异有统计学意义（P〈0.05）。结论：泽泻汤加味治疗梅尼埃病综合征,疗效满意,不良反应小。     

Implications d'ALK (anaplastic lymphoma kinase) en oncohématologie

The anaplastic lymphoma kinase gene (ALK) code for a receptor tyrosine-kinase. The fusion proteins from the ALK gene have been identified in oncohaematology malignancies including ALK positive anaplastic lymphoma large cell and non small cells lung cancer with EML4-ALK fusion gene. Constitutive activation generated by modification of this protein leads activation of anti apoptotic and survey pathways that makes it a prime target for these 2 subtypes of disease. Strategies and therapeutic molecules targeting the fusion protein are under development and preliminary results are encouraging. Therefore the mapping of the tumors is essential to help provide treatment specific to each entity. The best example is the chronic myeloid leukemia and the discovery of the fusion gene bcr-abl and of imatinib.     

Marqueurs biologiques de résistance à l'hormonothérapie dans les cancers du sein

During the past decade, molecular signatures allowed a better classification of breast carcinoma and a better evaluation of their prognosis. However, we still need predictive factors of treatment and /or prognosis factors specific of each patient. Regarding hormonal therapy, expression of hormone receptors is essential, but not sufficient to accurately predict response to treatment for all patients. To date, numerous data have identified different pathways of resistance to hormonal treatment, associated with heterogeneity in response to long term treatment. The challenge remains to identify and to anticipate these changes for each patient.     

The role of T-agglutinin in the disappearance of erythrocytes artificially aged by desialylation

The chance discovery of two mouse strains, one with and one without a high presence of serum T-agglutinin, permitted the investigation of the role of this antibody in the disappearance of desialylated erythrocytes, which may be regarded as a model for ageing. The proportional relationship between the quantity of sialic acid removed and the diminution of half-life is not affected by the presence or absence of T-agglutinin. Opsonization by T-agglutinin would therefore appear to be an improbable mechanism. Other possible theories are discussed.     

The formation of actin-containing gels by extracts of bovine splenic nerve

Extracts prepared from bovine splenic nerve at low ionic strength were concentrated by vacuum dialysis. When incubated with magnesium, adenosine triphosphate and the calcium chelating agent ethyleneglycol-bis(β-amino-ethyl)N,N'-tetra-acetic acid at 37°C for 1 h, the concentrated extracts formed strong, invertible gels. Guanosine triphosphate but not adenyl methylene diphosphate could substitute for adenosine triphosphate. Calcium ions and KCl above 0.1 M inhibited gelation. Gelation did not occur at 0°C. Gelation was prevented by cytochalasin and phalloidin but not by colchicine, podophyllotoxin or vinblastine.Sodium dodecyl sulphate electrophoresis of proteins sedimented from the invertible gels showed polypeptides which co-migrated with actin, α and β tubulin, bovine plasma albumin and myosin. Amongst the other polypeptides there was one with a molecular weight of about 215,000 and a doublet corresponding in mobility with the microtubule accessory proteins of high molecular weight.It is concluded that extracts of splenic nerve are able to form gels stabilized by fibres of actin which might be crosslinked by the polypeptides of molecular weight 215,000. The origin of actin found in extracts of splenic nerve is discussed.