A prospective, randomized clinical trial of antiretroviral therapies on carotid wall thickness

Objective: This article compares the effects of initiating three contemporary antiretroviral therapy (ART) regimens on progression of carotid artery intima-media thickness (IMT) over 3 years. Design: Randomized clinical trial. Setting: Multicenter (26 institutions). Patients: ART-naive HIV-infected individuals (n ¼ 328) without known cardiovascular disease or diabetes mellitus. Intervention: Random assignment to tenofovir/emtricitabine along with atazanavir/ ritonavir (ATV/r), darunavir/ritonavir (DRV/r), or raltegravir (RAL). Main outcome measures: Right-sided carotid IMT was evaluated by B-mode ultrasonography before ART initiation, and then after 48, 96, and 144 weeks. Comparisons of yearly rates of change in carotid IMT used mixed-effects linear regression models that permitted not only evaluation of the effects of ART on carotid IMT progression but also how ART-associated changes in traditional risk factors, bilirubin, and markers of HIV infection were associated carotid IMT progression. Results: HIV-1 RNA suppression rates were high in all arms (>85%) over 144 weeks. Modest increases in triglycerides and non-high-density lipoprotein cholesterol levels were observed in the protease inhibitor-containing arms compared with decreases with RAL. In contrast, carotid IMT progressed more slowly on ATV/r [8.2, 95% confidence interval (5.6, 10.8) mm/year] than DRV/r [12.9 (10.3, 15.5) mm/year, P ¼ 0.013]; changes with RAL were intermediate [10.7 (9.2, 12.2) mm/year, P ¼ 0.15 vs. ATV/r; P ¼ 0.31 vs. DRV/r]. Bilirubin and non-high-density lipoprotein cholesterol levels appeared to influence carotid IMT progression rates. Conclusion: In ART-naive HIV-infected individuals at low cardiovascular disease risk, carotid IMT progressed more slowly in participants initiating ATV/r than those initiating DRV/r, with intermediate changes associated with RAL. This effect may be due, in part, to hyperbilirubinemia

γH2Ax Expression as a Potential Biomarker Differentiating between Low and High Grade Cervical Squamous Intraepithelial Lesions (SIL) and High Risk HPV Related SIL.

BackgroundγH2AX is a protein biomarker for double-stranded DNA breakage; its expression was studied in cervical squamous intraepithelial lesions and carcinomas.MethodsImmunostaining for phospho-γH2AX was performed in sections from histologically confirmed cervical SIL and carcinomas, as well as from normal cervices used as controls. In total, 275 cases were included in the study: 112 low grade SIL (LGSIL), 99 high grade SIL (HGSIL), 24 squamous cell carcinoma (SCC), 12 adenocarcinoma and 28 cervical specimens with no essential lesions. Correlation of histological grading, high risk vs. low risk HPV virus presence, activated vs. non-activated status (by high risk HPV mRNA expression) and γH2AX expression in both basal and surface segments of the squamous epithelium was performed.ResultsGradual increase of both basal and surface γH2AX expression was noted up from normal cervices to LGSIL harboring a low risk HPV type, to LGSIL harboring a high risk virus at a non-activated state (p<0.05). Thereafter, both basal and surface γH2AX expression dropped in LGSIL harboring a high risk virus at an activated state and in HGSIL.ConclusionsγH2AX could serve as a potential biomarker discriminating between LGSIL and HGSIL, as well as between LGSIL harboring high risk HPV at an activated state

γH2Ax Expression as a Potential Biomarker Differentiating between Low and High Grade Cervical Squamous Intraepithelial Lesions (SIL) and High Risk HPV Related SIL

<div><p>Background</p><p>γH2AX is a protein biomarker for double-stranded DNA breakage; its expression was studied in cervical squamous intraepithelial lesions and carcinomas.</p><p>Methods</p><p>Immunostaining for phospho-γH2AX was performed in sections from histologically confirmed cervical SIL and carcinomas, as well as from normal cervices used as controls. In total, 275 cases were included in the study: 112 low grade SIL (LGSIL), 99 high grade SIL (HGSIL), 24 squamous cell carcinoma (SCC), 12 adenocarcinoma and 28 cervical specimens with no essential lesions. Correlation of histological grading, high risk vs. low risk HPV virus presence, activated vs. non-activated status (by high risk HPV mRNA expression) and γH2AX expression in both basal and surface segments of the squamous epithelium was performed.</p><p>Results</p><p>Gradual increase of both basal and surface γH2AX expression was noted up from normal cervices to LGSIL harboring a low risk HPV type, to LGSIL harboring a high risk virus at a non-activated state (p<0.05). Thereafter, both basal and surface γH2AX expression dropped in LGSIL harboring a high risk virus at an activated state and in HGSIL.</p><p>Conclusions</p><p>γH2AX could serve as a potential biomarker discriminating between LGSIL and HGSIL, as well as between LGSIL harboring high risk HPV at an activated state.</p></div

γH2AX as a regulator of cellular responses to HPV infection.

<p>An ATM-dependent DNA damage response is induced by HPV; however it is unclear which set of the ATM factors provide necessary functions. Complex feedback loops induce the phosphorylation of H2AX, that is recruited to regions flanking sites of double strand breaks. Proteins known to directly bind to / interact with H2AX are marked in bold. γH2AX forms complexes with these proteins associated with the viral DNA in HPV positive cells. Many of these loops interact with the HPV genome amplification that occurs during cycle arrest in late S/G2.</p

Histologically confirmed low grade cases (LGSIL) where high risk (HR) HPV type infection (LGSIL-HR) was identified.

<p>Comparison between activated (mRNA positive) and non-activated cases (mRNA negative) for 102 subjects. Statistically significant association are flagged with an asterisk.</p

γH2AX expression (basal and surface segments of epithelium) across histologically confirmed lesions for all patients with available data.

<p>Statistically significant associations (p<0.05) by ANOVA are flagged.</p

γH2Ax Expression as a Potential Biomarker Differentiating between Low and High Grade Cervical Squamous Intraepithelial Lesions (SIL) and High Risk HPV Related SIL - Fig 3

<p>γ2ΗΑΧ immunostaining of histological sections from the following cases: control (a); HGSIL (b); LGSIL harboring High Risk HPV at an activated state (c); and LGSIL harboring High Risk HPV at a non activated state (d). In pictures c and d, arrows depict positive cell (nuclear staining) at basal (white arrow) and surface (black arrow) area of the epithelium. Objective magnifications appear on pictures.</p

Basal and surface γH2AX expression (% positivity in 100 assessed cells) according to histological grade.

<p>Basal and surface γH2AX expression (% positivity in 100 assessed cells) according to histological grade.</p

Cytological vs. histological classification for the studied samples.

<p>Rates are given in percentage (%) for the histological category.</p