Objectives
Simplified methods for virological monitoring in resource-limited settings are increasingly needed. We evaluated the performance of the VERSANT® HIV-1 RNA (kPCR) assay for the ...determination of HIV-1 viral load from dried blood spots (DBS). Assay sensitivity and correlation with plasma quantification values were assessed.
Methods
A total of 98 DBS were prepared from fresh blood samples of HIV-infected patients. DBS were kept at room temperature for 6 weeks or 7 months before processing while the corresponding plasma samples were stored at −80°C. DBS were first pre-treated in a special DBS buffer. The DBS extracts and the plasma samples were then purified and amplified using the VERSANT assay reagents.
Results
In the first series of tests, performed after 6 weeks of storage, there was good correlation between quantification of viral load in plasma and in DBS (r = 0.95, P < 0.001). The detection rate in DBS was 100% when plasma levels were >1000 copies/mL. The sensitivity and specificity of the DBS assay were 88.2% 95% confidence interval (CI) 79.4-93.6 and 69.2% (95% CI 42.0-87.4), respectively. Using the 5000 copies/mL threshold (defining virological failure in resource-limited settings), both positive and negative predictive values were high (95.2% and 87.5%, respectively). After 7 months of storage there was a modest decrease in the detection rate and less significant correlations for samples with HIV-RNA <5000 copies/mL.
Conclusions
Quantification of HIV-RNA from DBS by the VERSANT automated sample preparation and detection method can be used to diagnose virological failure in HIV-positive patients.
The administration of antiretroviral therapy to lactating women could represent a possible strategy to reduce postnatal HIV transmission. In this study, we assessed the effect of antiretroviral ...treatment on breast milk viral load and determined plasma and breast milk drug concentrations in pregnant women receiving highly active antiretroviral therapy (HAART).
We studied 40 women receiving zidovudine, lamivudine, and nevirapine from 28 weeks of gestation to 1 month postpartum (group A) and 40 untreated pregnant women (group B). Blood and breast milk samples were collected at delivery and 7 days postpartum.
Women in group A had received a median of 85 days of therapy before delivery. Median breast milk concentrations of nevirapine, lamivudine, and zidovudine were 0.6, 1.8, and 1.1 times, respectively, those in maternal plasma. HIV RNA levels in breast milk were significantly lower in group A than in group B (median of 2.3 vs. 3.4 log at delivery and 1.9 vs. 3.6 log at day 7; P < 0.001 for both comparisons).
Antiretroviral drugs administered during the last trimester of pregnancy and after delivery reach levels similar to or higher than plasma concentrations in breast milk and can significantly reduce HIV RNA levels. Our data support the potential role of maternal HAART prophylaxis in reducing the risk of breast-feeding-associated transmission.
Childhood obesity is one of the most serious public health chal-lenges of this century. Overweight and obese children are likely to stay obese into adulthood and more likely to develop ...non-communicable diseases like diabetes and cardiovascular diseases at a younger age. In the WHO European Region one child out of 3, is overweight or obese. Over 60% of children who are overweight before puberty will be overweight in early adulthood. Children and adolescents, aged 5-19 have shown rising obesity rates in almost all nations, including where the situation was far from alarming 40 years ago. Several nations have seen the prevalence almost double: Israel has gone from 5.8% in 1975 to 11.9% in 2016, Andorra from 6.2% to 12.8%, and Malta from 7.4% to 13.4%. Analyzing overweight and obesity, we can see that they follow similar trends and patterns. In 1975 the majority of European countries had a prevalence less than 10% and obesity less than 5%, while no European country had overweight prevalence higher than 30% and obe-sity higher than 10%. In 2016 the trend reversed, showing a worrying increase in the number of European countries with a high prevalence of overweight (over 30%) and obesity (over 10%) (Fig. 1)(29). Starting from the analysis of epidemiological data on obesity in the WHO European Region, the paper analyzes the adopted prevention programs in order to assess their effectiveness and figure out the best strategies to reduce the prevalence of overweight and obesity. The WHO European Childhood Obesity Surveillance Initiative reported that children tend to overeat and not to do enough physical exercise. Different preventive programs have identified different areas of action and corresponding measures: consumption of healthy foods, physical exercise, care before conception and during pregnancy, early childhood, school age children, weight management, monitoring and evaluation. Primary prevention is essential to reduce obesity incidence: it is easier to act on the adoption of healthy eating habits than intervene with diets on children who already have weight issues. Working on pre-vention programs represents an investment for the future of children's health. By simply acting on prevention, particularly on body weight reduction, it could be possible to tackle the spreading of correlated di-seases. Therefore, prevention programs ought to be prioritized priority at a national and international level.
To identify factors associated with detectable viral load and the emergence of drug resistance in a cohort of HIV-infected pregnant women in Malawi receiving antiretroviral combination regimens for ...the prevention of mother-to-infant transmission.
The study included 260 treatment-naive women who had received a three-drug nevirapine-based regimen from week 25 of gestational age until 6 months after delivery. HIV RNA was determined at month 6 and drug resistance was assessed if viral load was >50 copies/mL. Attendance at the scheduled follow-up visits was used as an indirect measure of treatment adherence.
The rate of detectable HIV RNA at 6 months was 9.6% (25/260). The only significant predictor of this occurrence was the presence of ≥1 missed visit during follow-up (P = 0.012). Resistance was assessed in 19 of these women: 7 (37%) had a wild-type virus and the other 12 (63%) had resistance-associated mutations (nucleoside reverse transcriptase inhibitor, 7/12; non-nucleoside reverse transcriptase inhibitor, 11/12). Three of 12 cases (25%) in which mutations were detected had a viral load <1000 copies/mL. The emergence of resistance was not correlated with the presence of baseline mutations in either plasma or archived DNA.
In this cohort of women, detectable HIV RNA 6 months post-partum was infrequent and associated with low adherence to the treatment programme. Mutations were present in 63% of the women with detectable viral load at 6 months who had samples available for resistance testing. The impact of resistance on treatment re-initiation in women discontinuing drugs after the risk of transmission has ceased can be limited.
The development of drugs that can be used as topical microbicides is currently recognized as a priority area of research.
A preclinical evaluation of the potential effectiveness of TMC120, a ...non-nucleoside reverse transcriptase inhibitor (NNRTI), as a topical microbicide to prevent vaginal HIV-1 transmission in a humanized severe combined immunodeficient (hu-SCID) mouse model.
Reconstituted mice received an intravaginal application of a TMC120-containing gel 20 min prior to a non-invasive vaginal challenge with cell-associated HIV. The possible cytotoxic effect of TMC120-containing-gel on lymphocytes was assessed and their in vivo migration was followed using fluorescently labelled human lymphocytes. Systemic infection was monitored by p24 antigen detection in culture supernatant from cocultured intraperitoneal cells using antigen capture enzyme-linked immunosorbent assay test and by the presence of integrated proviral HIV-1 DNA in DNA extracted from spleen cells. In vivo migration of labelled lymphocytes was examined by analysis of cells isolated from regional lymph nodes.
In this model, systemic infection was successfully inhibited by the presence of TMC120-containing gel at vaginal level. The in vivo migration of human lymphocytes from the vagina to regional lymph nodes, following the deposition of TMC120-containing gel, excluded the possibility that inhibition of systemic infection was a result of NNRTI toxicity.
Vaginal transmission of HIV was successfully prevented by the application of a gel formulation containing TMC120. This is the first evidence of the in vivo effectiveness of a microbicide preparation containing an NNRTI against cell-associated HIV.
Objectives
The aim of the study was to determine the modifications of the mutational archive in proviral HIV‐1 DNA occurring during 24 months of intermittent or continuous highly active ...antiretroviral therapy (HAART).
Methods
The study population included subjects enrolled in the Istituto Superiore di Sanità Pulsed Antiretroviral Therapy (ISS PART) clinical trial. All of these patients were on first‐line HAART and had plasma HIV‐1 RNA below 50 HIV‐1 RNA copies/mL. A genotypic resistance test was performed on HIV‐1 DNA extracted from peripheral blood mononuclear cells (PBMC) at baseline and after 24 months of follow‐up. Resistance‐associated mutations (RAMs) were defined according to the International AIDS Society (IAS) USA classification.
Results
Sixty‐nine subjects were included in the study 36 enrolled in arm A of the ISS PART (continuous HAART) and 33 enrolled in arm B (intermittent HAART). No major modifications of the mutational archive were found in either group after 24 months of follow‐up, in terms of both the proportion of subjects with mutations and the total number of mutations.
Conclusions
In this patient population, the mutational archive in HIV‐1 DNA extracted from PBMC was stable for 24 months, irrespective of HAART modality, whether continuous or intermittent.
Purpose
To evaluate associations between CD4/CD8 ratio and pregnancy outcomes in women with HIV.
Methods
We evaluated, in a national study of pregnant women with HIV receiving antiretroviral ...treatment (ART), values of CD4/CD8 ratio at entry in pregnancy, changes between first and third trimester, and possible associations with preterm delivery, low birthweight, and HIV-RNA < 50 copies/ml at third trimester in univariate and multivariate analyses.
Results
Among 934 women, 536 (57.4%) were already on ART at conception. CD4/CD8 ratio (baseline value 0.570) increased significantly between the first and third trimesters, particularly in women who started ART in pregnancy (+ 0.163, vs. + 0.036 in women already on treatment). The rate of CD4/CD8 ratio normalization, defined by achieving a ratio ≥ 1 at the third trimester, was 13.2%. In multivariable analyses, women who entered pregnancy with a CD4/CD8 ratio < 0.3, compared to women with ratio ≥ 1, were almost four-times less likely to have third-trimester HIV-RNA < 50 copies/ml (AOR 0.258, 95%CI 0.111–0.601), and more than twice as likely to have preterm delivery (AOR 2.379, 95%CI 1.082–5.232). For preterm delivery, also a baseline CD4/CD8 ratio between 0.3 and 0.45 was significantly associated with an increased risk (AOR: 3.415, 95%CI 1.690–6.900).
Conclusion
We described for the first time independent associations of low CD4/CD8 ratio with preterm delivery and HIV-RNA suppression.
Abstract Background and aims Meta-analyses of randomized control trials investigating the association between incident diabetes and statin use showed an increased risk of new-onset diabetes (NOD) ...from 9% to 13% associated with statins. However, short follow-up period, unpowered sample size, and lack of pre-specified diagnostic criteria for diabetes detection could be responsible of an underestimation of this risk. We conducted a meta-analysis of published observational studies to evaluate the association between statins use and risk of NOD. Methods and results PubMed, EMBASE and MEDLINE databases were searched from inception to June 30, 2016 for cohort and case–control studies with risk of NOD in users vs nonusers, on ≥1000 subjects followed-up for ≥1 year. Two review authors assessed study eligibility and risk of bias and undertook data extraction independently. Pooled estimates were calculated by a random-effects model and between-study heterogeneity was tested and measured by I2 index. Furthermore, stratified analyses and the evaluation of publication bias were performed. Finally, the meta-analysis included 20 studies, 18 cohort and 2 case–control studies. Overall, NOD risk was higher in statin users than nonusers (RR 1.44; 95% CI 1.31–1.58). High between-study heterogeneity (I2 = 97%) was found. Estimates for all single statins showed a class effect, from rosuvastatin (RR 1.61; 1.30–1.98) to simvastatin (RR 1.38; 1.19–1.61). Conclusions The present meta-analysis confirms and reinforces the evidence of a diabetogenic effect by statins utilization. These observations confirm the need of a rigorous monitoring of patients taking statins, in particular pre-diabetic patients or patients presenting with established risk factors for diabetes.
Objective
We used data from a national study of pregnant women with HIV to evaluate the prevalence of congenital abnormalities in newborns from women with HIV infection.
Design
Observational study.
...Setting
University and hospital clinics.
Population
Pregnant women with HIV exposed to antiretroviral treatment at any time during pregnancy.
Methods
The total prevalence of birth defects was assessed on live births, stillbirths, and elective terminations for fetal anomaly. The associations between potentially predictive variables and the occurrence of birth defects were expressed as odds ratios (ORs) with 95% confidence intervals (95% CIs) for exposed versus unexposed cases, calculated in univariate and multivariate logistic regression analyses.
Main outcome measures
Birth defects, defined according to the Antiretroviral Pregnancy Registry criteria.
Results
A total of 1257 pregnancies with exposure at any time to antiretroviral therapy were evaluated. Forty‐two cases with major defects were observed. The total prevalence was 3.2% (95% CI 1.9–4.5) for exposure to any antiretroviral drug during the first trimester (23 cases with defects) and 3.4% (95% CI 1.9–4.9) for no antiretroviral exposure during the first trimester (19 cases). No associations were found between major birth defects and first‐trimester exposure to any antiretroviral treatment (OR 0.94, 95% CI 0.51–1.75), main drug classes (nucleoside reverse transcriptase inhibitors, OR 0.95, 95% CI 0.51–1.76; non‐nucleoside reverse transcriptase inhibitors, OR 1.20, 95% CI 0.56–2.55; protease inhibitors, OR 0.92, 95% CI 0.43–1.95), and individual drugs, including efavirenz (prevalence for efavirenz, 2.5%).
Conclusions
This study adds further support to the assumption that first‐trimester exposure to antiretroviral treatment does not increase the risk of congenital abnormalities.
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