ABSTRACT
PURPOSE: We have developed a sequencing assay for determining the usage of the genotypic HIV-1 co-receptor using peripheral blood mononuclear cell (PBMC) DNA in virologically suppressed HIV-1 infected patients. Our specific aims were to (1) evaluate the efficiency of V3 sequences in B versus non-B subtypes, (2) compare the efficiency of V3 sequences and tropism prediction using whole blood and PBMCs for DNA extraction, (3) compare the efficiency of V3 sequences and tropism prediction using a single versus a triplicate round of amplification. RESULTS: The overall rate of successful V3 sequences ranged from 100 % in samples with >3,000 copies HIV-1 DNA/10(6) PBMCs to 60 % in samples with <100 copies total HIV-1 DNA /10(6) PBMCs. Analysis of 143 paired PBMCs and whole-blood samples showed successful V3 sequences rates of 77.6 % for PBMCs and 83.9 % for whole blood. These rates are in agreement with the tropism prediction obtained using the geno2pheno co-receptor algorithm, namely, 92.1 % with a false-positive rate (FPR) of 10 or 20 % and of 96.5 % with an FPR of 5.75 %. The agreement between tropism prediction values using single versus triplicate amplification was 98.2 % (56/57) of patients using an FPR of 20 % and 92.9 % (53/57) using an FPR of 10 or 5.75 %. For 63.0 % (36/57) of patients, the FPR obtained via the single amplification procedure was superimposable to all three FPRs obtained by triplicate amplification. CONCLUSIONS: Our results show the feasibility and consistency of genotypic testing on HIV-1 DNA tropism, supporting its possible use for selecting patients with suppressed plasma HIV-1 RNA as candidates for CCR5-antagonist treatment. The high agreement between tropism prediction by single and triple amplification does not support the use of triplicate amplification in clinical practice.
Subject(s)
Genotyping Techniques/methods , HIV Infections/virology , HIV-1/genetics , HIV-1/physiology , Molecular Diagnostic Techniques/methods , Receptors, HIV/metabolism , Viral Tropism , Adult , DNA, Viral/chemistry , DNA, Viral/genetics , DNA, Viral/isolation & purification , Female , HIV Infections/diagnosis , HIV-1/classification , HIV-1/isolation & purification , Humans , Male , Middle Aged , Proviruses/classification , Proviruses/genetics , Proviruses/isolation & purification , Sequence Analysis, DNA , Virus InternalizationABSTRACT
UNLABELLED: Recently a bio-artificial liver (BAL) system has been developed at the Academic Medical Center (AMC) of Amsterdam to bridge patients with acute liver failure (ALF) to orthotopic liver transplantation (OLT). After successful testing of the AMC-BAL in rodents and pigs with ALF, a phase I study in ALF patients waiting for (OLT) was started in Italy. We present the safety outcome of the first 7 patients aged 21-56 years with coma grade III or IV The total AMC-BAL treatment time ranged from 8 to 35 hours. Three patients received 2 treatments with two different BAL's within three days. Six of the 7 patients were successfully bridged to OLT. One patient showed improved liver function after two treatments and did not need OLT. No severe adverse events of the BAL treatment were noted. CONCLUSION: Treatment of ALF patients with the AMC-BAL is a safe and feasible technique to bridge the waiting time for an adequate liver-graft.
Subject(s)
Liver Failure, Acute/therapy , Liver, Artificial , Adult , Extracorporeal Circulation , Female , Humans , Liver Transplantation , Liver, Artificial/adverse effects , Male , Middle Aged , Waiting ListsABSTRACT
The aim of this study was to evaluate efficacy of highly active antiretroviral therapy (HAART) in 64 HIV-1 positive treatment-naive and previously treated patients, with different viral load at baseline. The HAART regimen consisted of one protease inhibitor and two reverse-transcriptase inhibitors. Plasma RNA viral load was measured by RT-PCR (Roche Amplicor Monitor kit) at enrolment and at months 3, 6, 9, and 12 of follow-up. The viral load fell below the threshold of 200 copies/ml in 46.9% of patients at month 6 and this result lasted up to month 12 of follow-up in 42.2% of patients. In these patients the CD4+ cell count increased from a baseline with a median of 194 cells/mmc at month 12. Treatment failure occurred in 35.9% of patients and the proportion was higher among previously treated patients. 7 patients stopped therapy because of poor compliance and 5 because of adverse drug effects. Also in our cases the HAART regimen showed more efficacy in treatment-naive patients, whereas baseline viral load >1x105 was the cause of less efficacy of therapy.
ABSTRACT
200 mg UDPG or placebo respectively were administered after an observation period of 1 week to two randomly constructed groups of patients aged 15-35 yr with acute viral hepatitis but no prior history of disease. In the treated patients, mean decreases were most marked and most rapid in the case of SGOT, SGPT, alkaline phosphatase and total bilirubinaemia. These results offer good reason for supposing that UDPG can be usefully employed in the management of acute viral hepatitis.
