Immunoblot analysis for serodiagnosis of tuberculosis using a 45/47-kilodalton antigen complex of Mycobacterium tuberculosis.

We evaluated the immunoglobulin G (IgG) antibody response to the 45/47-kDa secreted protein of Mycobacterium tuberculosis by immunoblot assay, to assess its potential value for serological diagnosis. Control subjects consisted of healthy volunteers with negative or positive tuberculin skin tests. Most (>98%) scored negative in an immunoblot test when the sera were analyzed at a 1:400 dilution. Approximately 40% of sera (diluted 1 in 400) from tuberculous patients (positive smears) recognized the antigen complex. The sensitivity of the test for patients suffering from extrapulmonary tuberculosis was similar to that for patients suffering from pulmonary tuberculosis but who had negative smears. The frequency of positive reactions among the patients suffering from other pulmonary diseases was similar to that among the control subjects. In tuberculous patients infected with human immunodeficiency virus, the sensitivity of the immunoblot test was significantly lower. Thus, this test based on an antigen complex used in an immunoblot assay to detect the presence of IgG antibody has a specificity of 98% and a sensitivity of 40%. The simultaneous use of different purified antigens, selected at the same high specificity level, may improve the sensitivity of such an assay.     

Retinoids induce Fas(CD95) ligand cell surface expression via RARgamma and nur77 in T cells

Cells from the CD4+ murine T hybridoma line IP-12-7 enter the apoptotic suicide program via the Fas ligand (FasL)/Fas-mediated pathway upon TCR stimulation. This stimulus regulates the sensitization of the Fas death pathway and the cell surface appearance of preformed FasL. The apoptosis is dependent on new mRNA and protein synthesis and involves up-regulation of nur77.Two groups of nuclear receptors for retinoic acids (RA) have been identified: retinoic acid receptors (RAR) and retinoid X receptors. IP-12-7 cells express RARalpha and RARgamma. Here we show that,in the IP-12-7 T cells, RA also induced the expression and DNA binding of nur77, and the cell surface appearance of FasL. The induction was mediated via RARgamma. Despite the induced expression of cell surface FasL, only two structurally related RARgamma-selective compounds, CD437 and CD2325, initiated apoptosis in these cells. The lack of apoptosis induction by natural RA was related to the inability of RARgamma to sensitize the Fas death-pathway. Cell surface FasL, however, was able to induce cell death in Fas-bearing target cells. Natural RA also induced the expression of FasL in phytohemagglutinin-activated peripheral murine T cells. It is proposed that therapeutically administered RA might induce apoptosis in Fas-sensitive cells via induction of FasL expression in activated Tcells.     

Identical abnormality of the short arm of chromosome 18 in two Philadelphia-positive chronic myelocytic leukemia patients with erythroblastic transformation,resulting in duplication of BCR-ABL1 fusion

Two patients with Ph-positive chronic myelocytic leukemia in erythroblastic transformation and rearrangement of the short arm of chromosome 18 are reported. Fluorescence in situ hybridization studies showed that the 18p rearrangement resulted from translocation of the main part of chromosome 22 long arm to 18p, including BCR-ABL1 fusion. The 18p abnormality resulted, thus, in loss of 18p and duplication of BCR-ABL1 in both patients. The possible relation to the erythroblastic type of blastic phase is briefly discussed. In addition an apparently intact germline ABL1 gene was duplicated and inserted into chromosome 6 at band p21 in one of these patients.     

Noninvasive fluorescent study in situ and in real time of glucose effects on the pharmacokinetic of calcein

This study was undertaken to compare the effect of glucose injection on the pharmacokinetic behavior of a soluble dye in normal and tumoral tissues. The measurements were done using a noninvasive fluorescent spectroscopy in situ and in real time. The experiments were performed on three groups of animals with calcein as a soluble pH-insensitive fluorescent dye combined or not with glucose. Glucose solution was injected 5 or 30 min before calcein. Fluorescence emission intensity was recorded on normal and tumor tissues with an optical multichannel analyzer. Calcein concentration was also measured in blood using repetitive blood sampling. In the control group (without glucose injection), calcein is rapidly cleared from the blood, with a slow tissue clearance. Fluorescence of normal tissue was higher than fluorescence measured in tumor tissue. When glucose is injected 5 min before calcein, there was a rapid increase of tissue fluorescence followed by a plateau remaining during the whole experiment. No difference between tumor and normal tissue fluorescence intensity was observed. When glucose was injected 30 min before calcein, the plateau phase was reduced to 50 min in normal tissue. Tumor tissue fluorescence displays no distinct plateau phase. These results clearly showed the effect of glucose injection in situ and in real time, by a noninvasive method, on the pharmacokinetic of a soluble dye in a tumor tissue compared to a normal tissue. Differences between blood compartment and tissues kinetic profiles were also clearly demonstrated.     

Pre- and postnatal depression and coping: a comparative approach

BACKGROUND: The assessment of perinatal depressions and coping style. Methods: With depression scales (EPDS, GHQ.12) and Carver's BriefCope, the authors compared the prevalence rates of pre and postnatal depression in a cohort of 277 French women. RESULTS: Their work revealed very high levels of prenatal depression (almost 20%) and less important but nonetheless sizeable rates (11%) of postnatal depression, making such perinatal depressions a major public health concern. The coping styles proposed in Carver's public health BriefCope questionnaire make it possible to significantly differentiate during these two periods between depressive women and their non-depressed counterparts. CONCLUSIONS: This enables us to underline factors of risk and protection suggesting the importance of setting up compensatory and preventive systems and evaluating their pertinence in the framework of future research.     

Aggressive large granular lymphocyte lymphomas in five dogs: a clinical cytohistological and immunological study

Among 276 canine lymphomas referred to the Haematology–Cytology–Immunology laboratory of the Lyon Veterinary School between 1997 and 2003, there were five aggressive large granular lymphocyte (LGL) malignancies. The five dogs were clinically examined and followed up. Cytological and histological analyses of the liver, spleen, lymph nodes, intestine and bone marrow were performed. The immunophenotype and proliferation index were established. The most significant clinical finding was that of an aggressive clinical course, the presence of hepatomegaly and/or splenomegaly, an abdominal lymphadenopathy, anaemia in four cases and blood and bone-marrow involvement in two cases. The cytological presentation was a diffuse infiltration of atypical, large granular lymphoid cells. Two cases were of the null type (CD3–, CD79a–, CD4–, CD8–), and three were of the T-cell type (CD3+, CD79a–, CD4–, CD8+). The proliferation index was high in all cases, with a median of 54.4%. The histological presentation of the null-type cases was an infiltration of the livers portal triads and the spleens red pulp. The T CD8+ cases showed two different patterns, characterised by infiltration: in the first, of all the intestines layers, the livers portal triads, the spleens red pulp and the lymph nodes and, in the second, infiltration of the livers sinusoids, the spleens red pulp. Although these aggressive LGL lymphomas are still poorly known, they may be compared to three types of human lymphoma: aggressive NK cell lymphoma/leukemia, hepatosplenic T-cell lymphoma and enteropathy-type T-cell lymphoma.     

Mycobacterium bovis BCG induces similar immune responses and protection by rectal and parenteral immunization routes

We compared cellular immune responses to rectal, subcutaneous, and intradermal administration of Mycobacterium bovis BCG for 5 to 20 weeks in mice, guinea pigs, and macaques. Strong lymphoproliferative responses were induced in spleen cells after in vitro stimulation with purified protein derivative in guinea pigs and macaques, whatever the route of immunization. Comparable high numbers of gamma interferon- and tumor necrosis factor alpha-producing cells were found in the spleen after rectal, subcutaneous, and intradermal immunization of mice and macaques. Similar levels of precursors of cytotoxic T lymphocytes specific for mycobacterial antigens were observed in mice for all immunization routes. In macaques, cytotoxic activity, determined only at the end of the experiment (20 weeks), was similar after rectal and intradermal immunization. Six months after immunization, rectal and subcutaneous routes induced in mice similar levels of protective immunity against challenge with a virulent Mycobacterium tuberculosis strain (H37Rv). Rectal immunization gave immune responses and protective capacity similar to those for parenteral immunization and seemed to be a promising new route of vaccination against tuberculosis; in our study, immunization via the rectal route never induced side effects associated with parenteral routes (axillary adenitis) and could also effectively reduce the risks of viral transmission associated with unsafe injections in the developing world.     

Transvaginal sonography and rectal endoscopic sonography for the assessment of pelvic endometriosis: a preliminary comparison

BACKGROUND: Endometriosis and possible rectal involvement are difficult to assess by physical examination. Previous studies have shown the diagnostic value of magnetic resonance imaging and rectal endoscopic sonography (RES) in this setting, but not that of transvaginal sonography (TVS). The aims of this study were to compare the accuracy of TVS and RES for the diagnosis of pelvic endometriosis, and to compare the results with histological findings. PATIENTS AND METHODS: In a prospective study, 30 consecutive patients referred with clinical signs of endometriosis underwent TVS and RES; the images were interpreted blindly with regard to physical findings. RESULTS: Endometriosis was confirmed histologically in 28 (93%) of the 30 patients. Endometriomas were also present in 67% of cases. For the diagnosis of uterosacral endometriosis, the sensitivity, specificity, and positive and negative predictive values of TVS and RES were 75 and 75%, 83 and 67, 95 and 90%, and 45 and 40% respectively. For the diagnosis of rectosigmoid endometriosis, the sensitivity, specificity, and positive and negative predictive values of TVS and RES were 95 and 82%, 100 and 88%, 100 and 95%, and 89 and 64% respectively. CONCLUSION: Despite the large proportion of our patients who had intestinal endometriosis, representing a possible source of bias, our results suggest that TVS is as efficient as RES for detecting posterior pelvic endometriosis and should therefore be used as the first-line examination.