Transmitted drug resistance, selection of resistance mutations and moderate antiretroviral efficacy in HIV-2: Analysis of the HIV-2 Belgium and Luxembourg database

BACKGROUND: Guidelines established for the treatment of HIV-1 infection and genotype interpretation do not apply for HIV-2. Data about antiretroviral (ARV) drug efficacy and resistance mutations is scarce. METHODS: Clinical data about HIV-2 infected patients in Belgium and Luxembourg were collected and the effect of ARV therapy on plasma viral load and CD4 counts were analysed. Viral RNA encoding for protease (PR) and reverse transcriptase (RT) from ARV-naive and treated patients were sequenced. RESULTS: Sixty-five HIV-2 infected patients were included in this cohort. Twenty patients were treated with 25 different ARV combinations in a total of 34 regimens and six months after the start of ARV therapy, only one third achieved viral load suppression. All of these successful regimens bar one contained protease inhibitors (PIs). Mean CD4 gains in the group of viral load suppressors and the group of patients treated with PI-containing regimens were respectively significantly higher than in the group of non-suppressors and the group of PI-sparing regimens. The most frequent mutations selected under therapy (compared to HIV-2 ROD) were V71I, L90M and I89V within PR. Within RT, they were M184V, Q151M, V111I and K65R. All of these mutations, except K65R and M184V, were also found in variable proportions in ARV-naive patients. CONCLUSION: Despite a high rate of ARV treatment failure, better virological and immunological results were achieved with PI-containing regimens. The analysis of polymorphic positions and HIV-2 specific mutations selected during therapy showed for the first time that transmission of drug resistant viruses has occurred in Belgium and Luxembourg. The high heterogeneity in ARV combinations reflects a lack of guidelines for the treatment of HIV-2 infection

Breast cancer screening programmes--results of studies in foreign countries--situation in Belgium.

Results of randomized trials and case-control studies on breast cancer screening are reviewed. A combined analysis of data from the randomized studies indicates that a mortality reduction (29%) can be achieved by mammographic screening in women. Regular breast cancer screening by mammography for women aged 50-69 years reduces breast cancer mortality by 40% (confidence interval [CI] 95% about 27-55) in women who are screened at least once. In younger women, the results are inconclusive. The programme Europe against Cancer has recommended the setting-up of a network of pilot studies on breast cancer screening by mammography in the European Community in order to obtain the necessary experience, with a view to implementation of a national screening programme before the year 2000. In the Brussels project, the main objective is to develop a programme of quality assurance. In Belgium, there are other projects with different designs, for which the results are not yet available

Enregistrement du depistage des cancers dans la population. Enfin legal!

As a consequence of the privacy law of 8th December 1992, the registration of medical personal data was not permitted without the written consent of the person concerned. This law became an impediment for the elaboration of a screening register in the framework of population research for breast and cervical cancer. Even conclusive data protection measures were not taken into account as an exemption to this law. On 11th December 1998 this privacy law was adapted. Population research was included as an exeption on the ban on processing data that concern the health of natural persons. This clears the way for a Flemish screening registry for breast and cervical cancer within a legal framework.SCOPUS: ar.jinfo:eu-repo/semantics/publishe

Purification of human milk fucosyltransferase

The enzyme has been purified from skimmed milk concentrated on an Amicon PM 10 membrane and desalted on a Sephadex G 25 column equilibrated with 0.02 M MOPS buffer pH 7.4 contaning 10 mM MgCl2. This 2.5 fold concentrated preparation has been purified by affinity chromatography on a GDP Sepharose 4B column according to Barker et al. The fucosyltransferase active fractions were eluted by a 0.02 M MOPS buffer pH 7.4 containing 0.3 M lactose and 0.02 M EDTA. More than 99.4% of total proteins were eliminated in one step by this procedure. Two activity peaks were detected. This suggests the existence of different lactose: fucosyltransferases. In a second method, the concentrated human milk serum has been submitted to chromatography on DEAE cellulose suspended in 0.025 M phosphate buffer pH 6.9 and eluted with the same buffer. The first fractions contained maximal fucosyltransferase activity and were submitted to affinity chromatography. In this case, only one peak of fucosyltransferase activity has been detected. Acrylamide gel electrophoresis in the presence of sodium dodecylsulphate has been performed at each step of purification.SCOPUS: ar.jinfo:eu-repo/semantics/publishe

Specific and quantitative detection of human polyomaviruses BKV and JCV by LightCycler real-time PCR.

BACKGROUND: BK virus (BKV) and JC virus (JCV) are the only two known human polyomavirus that typically establish subclinical persistent infections. In immunocompromised hosts reactivation of the JCV infection is the cause of the central nervous system disease progressive multifocal leucoencephalopathy (PML), while BKV may cause renal nephropathy and haemorrhagic cystitis. OBJECTIVES: The goal of this study was to develop a specific quantification assay for each polyomavirus by LightCycler real-time polymerase chain reaction (PCR) based on SYBR Green I detection. STUDY DESIGN: DNA fragments of 138bp and 233bp from the "large T antigen" region of JCV and BKV, respectively, were amplified. The ability of the designed primer sets to separately quantify BKV DNA or JCV DNA and the PCR efficiency were assessed on reference DNA samples. Known amounts of cloned JCV DNA and BKV DNA from TEBU-BIO nv (Boechout, Belgium) were used to generate standard curves for the quantification assays. Species-specificity of the PCR was evaluated with cloned DNA and with DNA from patient samples. RESULTS: The assay allowed a specific quantification over a 7log dynamic range. Seventeen copies each of the viral genes were reproducibly and accurately detected. The primer sets generated specific DNA fragments for each virus confirmed by agarose gel analysis and by cycle sequencing. The similarities of the amplified gene sequences by BLAST analysis were 99% and 100% for BKV and JCV, respectively. There was no cross-reactivity within the dynamic range of the standard dilutions. CONCLUSIONS: We developed LightCycler real-time PCR assay based on SYBR Green I detection that provided rapid and specific quantification of polyomavirus load

Evaluation of a pilot quality assurance programme for breast cancer screening in the Brussels area

Background: a pilot quality assurance project for breast cancer screening was set up in the Brussels area in 1994. This paper aims to assess the performance of this programme after 4 years of activity, and the specific impact of consensual double reading of mammograms. Methods: each screening mammogram of women aged 50-69 year was submitted to a consensual double reading. Results of readings were registered with standardised forms. Follow-up data were traced for every positive mammogram. Results: 15.624 mammograms were performed in 12.239 women; recall rate at first round was 7,8%, open biopsy rate was 1%, cancer detection rate was 5,8%, positive predictive value of biopsy recommendation was 53,4%, benign to malignant biopsy ratio was 0,87:1, small size (less or equal to 10 mm) cancer proportion was 40%, proportion of cancers free of nodal involvement was 65%. Double reading yielded a 6% gain in sensitivity, while recall rate dropped from 8,1% to 7,8%. Conclusion: apart from a too high recall rate, screening performance was comparable with other published results in the same context; performance indicators ranged within norms recommended by "Europe against cancer". However, impact of double reading was weak and should be re-evaluated in the future perspective of a larger scale organised programme.SCOPUS: ar.jinfo:eu-repo/semantics/publishe

SE plots of the CT coding nucleotides.

<p>We created a SE plot for all the nucleotides (A) or for each nucleotide by codon (B). In graph A and B, the line represents moving averages (30 positions for total sequence in black; 10 positions for the separated nucleotides): blue for the first nucleotides of each codon in the Env reading frame (<i>env</i> N1), red for the second nucleotides (<i>env</i> N2) and green for the third nucleotides (<i>env</i> N3). The N1 variability has a major impact on Env and Rev and a small impact on Tat/Nef, the N2 variability has a major impact on Env and Tat/Nef and a small impact on Rev and finally the <i>env</i> N3 variability has a major impact on Tat/Nef and Rev and a small impact on Env. Above each plot are shown key regions of the different proteins. A: The first conserved region containing the endocytosis signal. B: The region containing the Kennedy-like sequences. C: The hydrophobic region. D: LLP-2 region. E: LLP-3 region. F: LLP-3 / LLP-1 interegion. G: LLP-1 region + position 166 for the stop codon.</p

Normalised SE plot of the CT by HIV-2 group.

<p>19 sequences from group A and 8 from group B are included. By contrast to the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0079129#pone-0079129-g002" target="_blank">Figure 2</a>, the SE are normalized by the natural log of the number of sequences to remove the entropy factor due to the sequences number. The lines represent the moving average of 10 positions (blue: group A, red: group B). The colours and letters below the plot represent the different studied regions: A: The first conserved region containing the endocytosis signal, B: The region containing the Kennedy-like sequences, C: The hydrophobic region, D: LLP-2 region, E: LLP-3 region, F: LLP-3 / LLP-1 inter-region, G: LLP-1 region + position 166 for the stop codon. </p