Exclusive TCRVbeta chain usage of ex vivo generated minor Histocompatibility antigen HA-1 specific cytotoxic T cells: implications for monitoring of immunotherapy of leukemia by TCRBV spectratyping

Tissue expression of minor Histocompatibility antigens HA-1 and HA-2 is limited to the hematopoietic system. Therefore, ex vivo generated HA-1/HA-2 specific cytotoxic T lymphocytes (CTLs) can be applied for adoptive immunotherapy of relapsed leukemia after HLA-matched HA-1/HA-2 mismatched stem cell transplantation. Here we used T cell receptor beta variable chain (TCRBV) spectratyping and/or TCRBV sequencing to monitor the specific TCR usage in eleven HA-1/HA-2 CTLs that were induced ex vivo with peptide pulsed dendritic cells. The HA-2 induced CTLs used different TCRBV. In contrast, the development of HA-1 specific CTLs coincided with prominent skewing of TCRBV7 spectratypes. Sequencing of the TCRBV7 specific PCR products used by these ex vivo generated HA-1 CTLs revealed the exclusive usage of TCRBV7-9*03, identical to the TCRBV used by HA-1 specific CTLs induced in vivo after stem cell transplantation. Thus, monitoring of immunotherapy with HA-1 specific CTLs is now also feasible by TCRBV spectratyping.     

Slug species- and population-specific effects on the end points of the Slug Mucosal Irritation test.

The Slug Mucosal Irritation test seems to be a promising method for the evaluation of the local tolerance of products applied to the mucosa. Furthermore, the Slug Mucosal Irritation test is a reliable method to classify chemicals accurately into three eye irritation categories based on the mucus production and the score for tissue damage. Until now the slug Arion lusitanicus collected in Belgium was always used as test organism. The present study investigated the effects of the slug population and species on the end points of the test and on the eye irritation classification. For this purpose, the eye irritation/damage test procedure and eye reference chemicals were used. Comparison of the results of one Belgian and two Swiss A. lusitanicus populations indicated that the geographic and ecological origins of slug populations did neither influence the mucus production, nor the score for tissue damage. Slug species-specific effects on the test end points were investigated by comparing the data of Belgian Limax flavus with corresponding data of Belgian A. lusitanicus. L. flavus produced more mucus than A. lusitanicus, so that the mucus production cut-off values had to be increased. Therefore, the results indicated that the test procedure and prediction model have to be optimised and validated, if other slug species are used instead of A. lusitanicus.     

Female genital mutilation: knowledge, attitudes and practices of Flemish gynaecologists.

ObJECTIVE: To assess the knowledge, attitudes and practices with regard to female genital mutilation (FGM) among gynaecologists in Flanders, Belgium. METHODS: A questionnaire-based survey was sent to 724 Flemish gynaecologists and trainees. RESULTS: Three-hundred-and-thirty-four questionnaires were returned. The survey revealed gaps in the knowledge of FGM and the provision of care by Flemish gynaecologists to women who had been mutilated. It also appeared that FGM was not properly addressed in the basic and specialized medical training in Flanders, that little was known about codes of conduct issued by the hospitals when these were not lacking altogether, and that knowledge about legislation concerning FGM was deficient. There was much confusion whether re-infibulation is authorized, and what its legal status is. Some respondents considered cosmetic vaginal surgery as a form of FGM and many were in favour of the medicalization of FGM. Gynaecologists were most commonly confronted with complaints related to sexual problems caused by FGM. Finally, the study also showed that only about a third of the gynaecologists were discouraging women from having their daughters excised. CONCLUSION: There is a need for a thorough discussion among all those concerned of the ethical and legal aspects of re-infibulation, medicalization of FGM and cosmetic vaginal surgery.     

The Significance of Recurrent Polyhydramnios

Summary: A study of 30 women who had polyhydramnios in more than 1 pregnancy revealed that 8 of the 36 resultant pregnancies (22.2%) were associated with diabetes mellitus, 14 (37.8%) with fetal macrosomia, and the perinatal mortality rate was 16.2% (6 of 37). The incidence of major fetal malformations or abnormalities was 18.9% (7 of 37); 4 of the 6 deaths resulted from malformations (anencephalus (2), hydrocephalus (1), nonimmune hydrops (1)), and the other 2 deaths were from hyaline membrane disease associated with prematurity. Recurrent polyhydramnios occurred in 1 in 1,720 pregnancies. The onset was acute in 3, subacute in 2 and chronic in 31, the perinatal deaths in these categories being 2, 1 and 3 respectively. The risk of recurrent polyhydramnios is the risk of fetal malformation and premature delivery. The latter may be preventable by prompt therapy with indomethacin, and serial amniocentesis if this therapy fails.     

Enhanced expression of IFN-gamma mRNA in CD4(+)or CD8(+)tumour-infiltrating lymphocytes compared to peripheral lymphocytes in patients with renal cell cancer

The mRNA expression of the cytokines IFN-gamma, IL-10 and TNF-alpha and the proapoptotic factor Fas ligand (FasL) was compared in freshly isolated CD4(+)and CD8(+)tumour-infiltrating lymphocytes (TIL) and simultaneously obtained autologous CD4(+)and CD8(+)peripheral blood lymphocytes (PBL) from 20 patients with renal cell carcinomas (RCC). TIL were isolated from mechanically disaggregated tumour material and PBL from peripheral blood by gradient centrifugation. The cells of the interphase were depleted from tumour cells with anti-human epithelial antigen magnetic beads and then positive selection was performed with anti-CD4 or anti-CD8 magnetic beads. In these pure lymphocyte preparations the constitutive expression of cytokine and FasL mRNAs was determined by using a PCR-assisted mRNA amplification assay. In the CD4(+)TIL from the 20 patients with RCC, levels of mRNAs encoding for IFN-gamma (P相似文献   

Absence of behavioral deficits following neonatal undernutrition in the rat

Previous studies of the effects of early undernutrition on behavior in adult rats have confounded underfeeding with maternal deprivation or membership in a large litter. In the present experiment food-deprived rats received full-time maternal care and lived in the same-sized litters as well-fed controls. In contrast to previous findings food-deprived animals in the present study did not differ from controls in the open-field test, in a test of motor coordination, and in 2 learning tasks. However, food-deprived animals were more active than controls in a residential plus maze. Females showed less effect of food deprivation on body growth, but a much greater effect on activity, than males. These findings suggest that early undernutrition when not confounded with social and maternal deprivation may have more restricted effects on adult behavior than has been previously believed.     

Nebulization of four commercially available amphotericin B formulations in persistently granulocytopenic rats with invasive pulmonary aspergillosis: evidence for long-term biological activity

The nebulization of amphotericin B desoxycholate (AMB-DOC), liposomal amphotericin B (L-AMB), amphotericin B lipid complex (ABLC) and amphotericin B colloidal dispersion (ABCD) has been investigated. Particle sizes of generated aerosol droplets were measured. Pulmonary amphotericin B deposition and amphotericin B concentration in blood directly after nebulization and at six weeks after nebulization was measured in healthy rats. The efficacy of nebulized amphotericin B formulations was evaluated in persistently granulocytopenic rats with invasive pulmonary aspergillosis. Treatment was given either after or before fungal inoculation. The endpoint was survival of animals. Aerosol particle sizes, expressed as the values for the mass median diameter were 1.38, 2.43, 0.90 and 2.29 microm for AMB-DOC, L-AMB, ABLC and ABCD, respectively. Amphotericin B concentrations in the lungs directly after nebulization exceeded the minimum inhibitory concentration of Aspergillus fumigatus and amphotericin B was still detected in lungs of rats at six weeks after nebulization. Treatment, started at 16 h after fungal inoculation, resulted in a significantly prolonged survival as compared with sham-treated rats for all four formulations. Prophylactic treatment at one week before fungal inoculation resulted in a significantly prolonged survival for all four formulations. Aerosol treatment given at two weeks before inoculation was effective only for AMB-DOC and L-AMB, whereas treatment given at six weeks resulted in a significantly prolonged survival for L-AMB only. All commercially available amphotericin B preparations could be nebulized efficiently and may be of value in the prophylactic treatment of invasive pulmonary aspergillosis.     

Modelling the association between adherence and viral load in HIV-infected patients

The primary objective of this paper is to investigate the effect of adherence to prescribed antiretroviral therapy on virologic response measured repeatedly over time in HIV-infected patients. To this end observations on plasma viral load (HIV RNA) assessed in copies/ml are categorized into four clinically meaningful states, [0--50[, [50--400[, [400--2000[, [2000 and up. A time-dependent continuation ratio model is used to analyse longitudinal ordinal responses. The main challenge lies in modelling dependencies over time and using information contained in the data efficiently to establish a dynamic relation between observed patient adherence and viral load. Among the several measures of adherence investigated, two specifically account for long periods of time without intake. One is derived from the third moment of the inter-dose interval distribution, while the second reflects internal drug exposure using pharmacokinetic parameters. The approach is applied to a clinical trial involving 35 patients who were followed over 12 months. Results demonstrate a significant relation between patient adherence and virologic response.