The design, development and clinical success of HIV protease inhibitors represent one of the most remarkable achievements of molecular medicine. This review describes all nine currently available FDA-approved protease inhibitors, discusses their pharmacokinetic properties, off-target activities, side-effects, and resistance profiles. The compounds in the various stages of clinical development are also introduced, as well as alternative approaches, aiming at other functional domains of HIV PR. The potential of these novel compounds to open new way to the rational drug design of human viruses is critically assessed.

Human immunodeficiency virus type 1 (HIV-1) latency represents the major barrier to virus eradication in infected individuals because cells harboring latent HIV-1 provirus are not affected by current antiretroviral therapy (ART). We previously demonstrated that DNA methylation of HIV-1 long terminal repeat (5’ LTR) restricts HIV-1 reactivation and, together with chromatin conformation, represents an important mechanism of HIV-1 latency maintenance. Here, we explored the new issue of temporal development of DNA methylation in latent HIV-1 5’ LTR. In the Jurkat CD4+ T cell model of latency, we showed that the stimulation of host cells contributed to de novo DNA methylation of the latent HIV-1 5’ LTR sequences. Consecutive stimulations of model CD4+ T cell line with TNF-α and PMA or with SAHA contributed to the progressive accumulation of 5’ LTR DNA methylation. Further, we showed that once established, the high DNA methylation level of the latent 5’ LTR in the cell line model was a stable epigenetic mark. Finally, we explored the development of 5’ LTR DNA methylation in the latent reservoir of HIV-1-infected individuals who were treated with ART. We detected low levels of 5’ LTR DNA methylation in the resting CD4+ T cells of the group of patients who were treated for up to 3 years. However, after long-term ART, we observed an accumulation of 5’ LTR DNA methylation in the latent reservoir. Importantly, within the latent reservoir of some long-term-treated individuals, we uncovered populations of proviral molecules with a high density of 5’ LTR CpG methylation. Our data showed the presence of 5’ LTR DNA methylation in the long-term reservoir of HIV-1-infected individuals and implied that the transient stimulation of cells harboring latent proviruses may contribute, at least in part, to the methylation of the HIV-1 promoter.

Objectives: The aim of this study was to evaluate the incidence and epidemiological characteristics of infective endocarditis (IE) in the Czech Republic. These results represent the first data on the epidemiology of IE from the post-communist countries. Methods:This was a prospective multi-centre observational study monitoring the occurrence of IE in the catchment areas of 29 hospitals during a 12-month period. The total monitored territory involved a population of 3.9 million people (37.7% of the total Czech population). Patients were included in the study if they had a diagnosis of possible or definite endocarditis according to the modified Duke criteria. Results: One hundred and thirty-four episodes of IE in 132 patients were reported. Thus the crude incidence of IE was 3.4 cases/100,000 inhabitants/y. Vegetations were most frequently found on the aortic and mitral valves. The most frequent agent was Staphylococcus aureus (29.9%). The aetiology remained unexplained in 33.6% of cases, mainly because of previous antibiotic therapy. Surgical intervention during antibiotic therapy was performed in 36 patients (27.5%). Thirty-six patients died during hospitalization (in-hospital mortality rate 27.5%). The most common predisposing cardiac factors were remote cardiac surgery (19.4%) and degenerative valvular changes (11.9%). The most common extracardiac factors were pyogenic infections of skin and soft tissues (13.0%) and chronic haemodialysis (8.2%). Conclusions: Our results document the changing epidemiological characteristics of IE, namely an increasing incidence of the disease and an increasing role of Staphylococcus aureus as a major pathogen. A shift was evident in predisposing factors for IE: almost 39% of IE episodes were associated with both cardiac and extracardiac modern medical procedures.

Depletion and functional impairment of circulating plasmacytoid dendritic cells (pDCs) are characteristic attributes of HIV-1-infection. The mechanism of dysfunction of pDCs is unclear. Here, we studied the development of phenotype of pDCs in a cohort of HIV-1-infected individuals monitored before the initiation and during a 9-month follow up with antiretroviral therapy (ART). Using polychromatic flow cytometry, we detected significantly higher pDC-surface expression of the HIV-1 receptor CD4, regulatory receptor BDCA-2, Fcγ receptor CD32, pDC dysfunction marker TIM-3, and the marker of killer pDC, TRAIL, in treatment-naïve HIV-1-infected individuals before initiation of ART when compared to healthy donors. After 9 months of ART, all of these markers approached but did not reach the expression levels observed in healthy donors. We found that the rate of decline in HIV-1 RNA level over the first 3 months of ART negatively correlated with the expression of TIM-3 on pDCs. We conclude that immunogenic phenotype of pDCs is not significantly restored after sustained suppression of HIV-1 RNA level in ART-treated patients and that the level of the TIM-3 expressed on pDCs in treatment naïve patients could be a predictive marker of the rate of decline in the HIV-1 RNA level during ART.