Introduction
Azole resistance in Aspergillus fumigatus is increasingly reported. We describe the validation of
AsperGenius® , a new multiplex real-time polymerase chain reaction (PCR) assay ...consisting of two multiplex real-time PCRs: one which identifies the clinically relevant Aspergillus species, and one which detects the TR34, L98H, T289A, Y121F mutations in CYP51A and differentiates susceptible from resistant A. fumigatus strains.
Methods
The diagnostic performance was tested on 37 bronchoalveolar lavage (BAL) samples from
haematology patients and on 40 BAL samples from intensive care unit (ICU) patients using BAL
galactomannan ≥1.0 or positive culture as the gold standard for the presence of Aspergillus.
Results
In the haematology and ICU groups combined, there were 22 BAL samples with IA (2 proven, 9
probable and 11 non-classifiable). Nineteen of the 22 BAL samples were positive according to the gold standard. The optimal cycle threshold value for the presence of Aspergillus was <36. Sixteen of the 19 BAL samples had a positive PCR (2 Aspergillus species and 14 A. fumigatus). This resulted in a sensitivity, specificity, positive and negative predictive value of 88.9%, 89.3%, 72.7% and 96.2% for the haematology group and 80.0%, 93.3%, 80.0% and 93.3% in the ICU group, respectively. The CYP51A real-time PCR confirmed 12 wildtype and 2 resistant strains (1 TR34/L98H and 1 TR46/Y121F/T289A mutant).
Conclusion
The AsperGenius® multiplex real-time PCR allows for a sensitive and fast detection of Aspergillus species directly in BAL samples. More importantly, this assay detects and differentiates wildtype from resistant strains even if BAL cultures remained negative.
Objectives: To investigate the epidemiological determinants responsible for the high number of anthropophilic dermatophytes received by the National Reference Center for Mycosis of Liege (NRCL) ...during the year 2012. To perform a genotypic characterization by the Diversilab® system focusing on the two main isolated species, Microsporum audouinii and Trichophyton violaceum. To present a preliminary study preceding the national survey launched in 2013.
Methods: A total of 51 strains of M. audouinii (50 clinical + 1 reference (ref.) strains) and 15 strains of T. violaceum (14 clinical + 1 ref. strain) originating from different locations through Belgium were included in the study. The fungal strains were first cultivated on Malt agar, then sub-cultured in Sabouraud liquid medium (Fluka). The grown mycelium was processed for DNA extraction following recommendations of the manufacturer (Ultra Clean® DNA Microbial isolation kit, MoBio laboratories). Genotypic analysis was performed using the DiversiLab® system (BioMérieux) for DNA fingerprinting and analysis.
Results: Regarding M. audouinii, four different genotypic groups of strains were separated. Group 1 includes 11 strains and is only found in the Liège surroundings. Group 2 includes only one strain with little differences compared to group 1 and collected from the Liège area. These two groups may be related to each other. Group 3 contains 36 strains and the reference strain. This genotype is distributed in different Belgium locations. The last group, group 4, contains only 3 isolates sharing low similarities in comparison with the 3 other groups. Concerning T. violaceum, 6 different genotypic groups with a mixed geographical distribution were determined. Group 1 includes 8 clinical isolates and the ref. strain. The other five isolates are all different and seem not to be related to each other.
Conclusion: The automated typing DiversiLab® system proved to be an easy and efficient method to investigate the molecular epidemiology of dermatophytes infections. Preliminary results of the study show that, through Belgium, several groups of isolates co-exist for M. audouinii and T. violaceum providing evidence of genetic heterogeneity. This variation can be related to acquired mutations due to environmental adaptation. Further investigations are necessary to better understand the impact of this genotypic variation.
The FLRG gene encodes a secreted glycoprotein that binds to activin and is highly homologous to follistatin, an activin ligand. We cloned the promoter region of the human FLRG gene, and defined the ...minimal region necessary for transcription activation in a reporter-system assay. We showed that the fragment between positions −130 and +6, which consists of multiple consensus Sp1-binding sites, is required for the constitutive expression of the FLRG gene. We demonstrate here that FLRG mRNA expression is rapidly induced by TGFβ or by transfection with Smad protein expression vectors in human HepG2 cells. We investigated the transcription-regulation mechanism of FLRG expression in HepG2 cells following treatment with TGFβ. By deletion and point-mutation analysis of the FLRG promoter, we identified a Smad-binding element involved in the TGFβ-inducible expression of the FLRG gene. Moreover, transactivation of the FLRG promoter by TGFβ was compromised by dominant-negative mutants of Smad3 and Smad4 proteins. In addition, gel electrophoresis mobility-shift assays demonstrated the specific interaction of Smad3 and Smad4 proteins with the Smad-binding element consensus motif found in the FLRG promoter. Taken together, our data imply that Smad proteins participate in the regulation of expression of FLRG, a new target of TGFβ transcription activation.
Summary
The translocation t(14;18) and its t(2;18) and t(18,22) variants, which involve the
BCL
2
genetic hallmark for follicular lymphoma (
FL
), have been reported in several cases of chronic
B
...‐cell lymphoproliferative disease (
CLPD
) and frequently in chronic lymphocytic leukaemia (
CLL
). We describe here the clinical, morphological, immunological, cytogenetic and molecular findings from 37 cases of t(14;18)‐positive
CLPD
, identified from our series of non‐
FL
B‐cell neoplasms (
n
= 993) that were routinely analysed in peripheral blood by conventional cytogenetics analyses. The
FL
diagnosis was excluded by morphology and immunology (the samples were
CD
10 negative in all cases). The
BCL
2
translocations were observed in 22
CLL
cases, including 7 monoclonal
B
‐cell lymphocytosis (
MBL
) cases re‐classified according to the new International Workshop on
CLL
criteria, six small lymphocytic lymphoma (
SLL
) cases, 1 splenic marginal zone lymphoma (
SMZL
) case and eight cases of unclassifiable
CLPD
with overlapping
CLL
/
MZL
features. In the
CLL
cases, the
IGH
/
BCL
2
fusion was remarkably associated with trisomy 12 (13/22) and mutated
IGHV
status (20/21) and did not affect the outcome. Moreover, most of these
CLL
s harboured a low mutation load of
BCL
6
gene and unmutated
FAS
(
CD
95
) loci, which points to a post–germinal‐centre cellular origin.
Abstract 4549
Patients with high risk AML have poor outcomes. However, the only approach with curative potential remains allogeneic HSCT. With the aim to improve the effect of allogeneic HSCT by ...sequential use of chemotherapy, RIC regimen and pDLI, we are currently conducting a prospective pilot study for high risk AML. High risk AML was defined by unfavorable cytogenetics, adverse molecular abnormality, secondary AML, and AML requiring 2 induction courses to obtain CR1.
After identification of a HLA 10/10 donor, patients are received the sequential regimen consisted of fludarabine (30mg/m2/d), Ara-c (2g/m2/d) and amsacrine (100mg/m2/d) (FLAMSA) chemotherapy for 4 days. After 3 days of rest, RIC regimen consisted of 4 Gy TBI, cyclophosphamide for 2 days (40 mg/kg in case of matched related donors, and 60 mg/kg for unrelated or mismatched donors), and ATG (5mg/kg total dose) (German regimen) or Busulfan 3.2mg/kg/d during 4 days followed by ATG (5mg/kg total dose) (French regimen). The modified regimen has been established after our results in refractory AML patients (ASH 2011, poster 1957). Prophylactic donor lymphocyte transfusion was given from day +120 in patients who were not receiving immunosuppression and were free of GvHD.
Our objective is to include 20 patients and to compare with a control cohort of patients with the same high risk AML treated according to the conventional strategy during the same period.
Between August 2010 and November 2011, we have included 12 consecutive patients in first complete response who underwent an allogeneic HSCT after sequential FLAMSA-RIC regimen with a median follow-up of 12 months (range 7–22). Nine patients were < 55 years old (median age: 54 28–64), 7 patients had an unrelated donor and 5 patients had a related donor. The stem cell source was PBSC for 11 patients and two cord blood unit for 1 patient. Before FLAMSA-RIC regimen, 3 patients had received two induction courses to obtain CR1. All patients had adverse cytogenetics or molecular abnormalities and 1 patient had a secondary AML.
At the last follow-up, 6 patients (50%) are alive in CR. (Figure 1.) Four patients (33.3%) died in remission. The cause of death was infection for 2 patients, aGvHD for 1 patient and graft failure for 1 patient. Only one patient died from relapse 6 months after transplantation.
Five patients (41.6%) experienced aGvHD and 2 patients (16.6%) had an extensive cGvHD including the patient who has been transplanted with 2 cord blood unit. Six patients (75%) in a group of 8 patients aged > 45 years experienced complications (infection (n=3) and GvHD (n=3)). One patient (25%) from a group of 4 patients aged < 45 years had infectious complication after transplantation. Prophylactic donor lymphocyte transfusion was given in 6 patients, the causes of no administration were GvHD for 2 patients, cord blood unit as stem cell source for 1 patient and 3 patients were dead before 120 days after transplantation. From the 6 patients who had received pDLI, 5 patients are alive in CR and 1 patient died from GvHD.
The FLAMSA-RIC regimen before allogeneic HSCT is a new approach for high risk AML. Between 2012 January and 2012 July, 8 additionnal patients have been included and the results for the whole study will be communicated later. Our primary results are promising especially for young patients (< 45 years) who seem to better profit from this sequential FLAMSA-RIC regimen. Display omitted
Nicolini:Novartis, Bristol Myers-Squibb, Pfizer, ARIAD, and Teva: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding.
Il s'agit d'une enquête réalisée à Amiens du 1er mars 1993 au 28 février 1994 chez les femmes venant d'accoucher pour évaluer leur niveau d'immunité vis à vis de la toxoplasmose et leur connaissances ...des mesures préventives. L'analyse des données s'est faite grâce au locigiel Epidemio sur un total de 987 femmes. La séroprévalence est de 58% et l'analyse montre que c'est la consommation de viande peu ou pas cuite et la présence d'un chat dans l'entourage qui sont les 2 facteurs de risque majeurs. La quasi totalité des femmes a pu citer au moins 2 moyens de prévention efficaces et les reccommandations sont largement suivies. En conclusion, les résultats sont rassurants. Cependant, une enquête nationale serait nécessaire pour permettre une évaluation sur le plan national.
Quantitative monitoring of imatinib mesylate (IM)-resistant, mutated BCR-ABL
+ cells during the follow-up of CML could be useful for optimizing therapeutic management. We retrospectively analyzed ...T315I mutated BCR-ABL clones throughout the CML history of two patients by nested-PCR-RFLP. At the time of progression, the T315I mutation represented 100% of the BCR-ABL transcripts. During follow-up, we showed that (i) despite a molecular response to IM, a high proportion of T315I transcripts were present (>85%) and predictive of relapse, (ii) interruption of IM and switching to other therapies resulted in a significant reduction in mutant transcript level while total BCR-ABL
+ transcripts remained stable.
A case of severe amebic colitis in a 21 year-old french patient is described. It raises the problems of modality and place of contamination,
Entamoeba histolytica levels of virulence and ...differenciation between
E. histolytica and
E. dispar.
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