Genotypic testing on HIV-1 DNA as a tool to assess HIV-1 co-receptor usage in clinical practice: results from the DIVA study group.

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.

Cost-benefit evaluation of influenza vaccination in the elderly in the Italian region of Liguria.

Influenza causes considerable morbidity and mortality and the damage to public health can be considerable. The most effective measures available for the prevention of influenza is vaccination. In most industrialised countries the objective of vaccination is to limit the disease among individuals at risk, especially the elderly. During the winter of 2000/2001, General Practitioners (GPs) monitored 14,818 elderly individuals. The objective was to evaluate the weekly incidence of the disease. Furthermore, we carried out a prospective study on 512 elderly individuals, arranged according to vaccination (304 vaccinated and 208 non-vaccinated), with the main objective of assessing the costs of the disease and the efficacy of vaccination. Finally, in order to assess the percentage of vaccinated elderly individuals, we carried out a telephone survey on 500 subjects. Our clinical surveillance study enabled us to establish that morbidity was particularly low in elderly individuals.The results of the prospective study allowed us to estimate the cost-benefit ratio at 8.22, with a net saving of 110.20 Euros for each vaccinated subject. We were also able to establish that the vaccine coverage among elderly individuals was 63%. Our study, though carried out during a low epidemic year, confirms the economic advantage of vaccination in the elderly.

[Chemico-experimental data on the principal bile constituents in patients with obstructive jaundice]. / Rilievi chimico-sperimentali sui principali costituenti biliari in pazienti con ittero ostruttivo.

Bile samples from patients suffering from cholestasis were tested. Cholesterol, phospholipids, and bile acids (cholic, lithocholic, deoxycholic, chenodeoxycholic) were measured and the methods for the gas-chromatographic determination of cholesterol and major bile acids as well as for the colorimetric determination of phosphorus in phospholipids of human bile are described in extenso. Bile samplings were first carried out on the day the drainage tube was positioned and were repeated every 5 days for four times. Between the first and the last sampling, 1250 mg of phosphatidylcholine was intravenously administered to each patient daily. The aim of the experiment was to evaluate the possible variations in the bile constituents occurring over the specified time.

Preparation of small volume, leuko and erythrocyte very poor platelet concentrates.

Recently developed automated discontinuous flow centrifuge (DFC) separators can produce leuko- and erythrocyte-poor platelet concentrates (PC). According to general experience with these machines it is difficult to obtain more than 4 X 10(11) platelets, though a second program set up by Coffe et al. appears to produce PC containing approximately 5 X 10(11) platelets suspended in a plasma volume of 390 ml. At our center we employed a new Dideco cell separator equipped with the surge pump and a technique developed for the production of small volume, RBC and WBC-very poor PC. In 60 routine procedures we obtained the following results: mean processing time 87 +/- 11 minutes; final volume of PC 136 +/- 19 ml, with a mean platelet yield of 5.21 X 10(11) platelets. WBC contamination was 1.8 X 10(8) (93% lymphocytes) and RBC were 3.1 X 10(8). Plasma volume as well as WBC and RBC contamination were reduced by recirculating PC after the 6th pass. The demand for single donor platelet concentrates (PC) is increasing progressively. Recently developed automated cell separators can produce leukocyte (WBC) and erythrocyte (RBC) poor PC. With these machines it may be difficult to obtain PC containing at least 4 X 10(11) platelets and less than 1 X 10(9) leukocytes (1, 2, 3) since donor variables such as hematocrit, precounts, buffy coat formation and initial plasma light transmission are of paramount importance for the efficiency of the program. At our center a prototype discontinuous flow centrifuge (DFC) cell separator equipped with the surge pump was studied.(ABSTRACT TRUNCATED AT 250 WORDS)