Switching virally suppressed, treatment-experienced patients to a raltegravir-containing regimen does not alter levels of HIV-1 DNA

Background

Current HIV-1 antiretroviral therapy (ART) greatly reduces virus replication but does not significantly affect the viral reservoir. Raltegravir, a recently introduced integrase inhibitor, could, at least theoretically, reduce residual viremia in patients on ART and affect the viral reservoir size. The aim of this study was to assess whether switching therapy in treatment-experienced patients that were virally suppressed to a raltegravir-containing regimen reduces the size of the viral reservoir, and if such treatment leads to a change in levels of HIV 2-LTR circles in this patient group.

Methods

14 ART experienced individuals with a suppressed viral load (<50 HIV-1 RNA copies/mL plasma) at baseline (for at least 2 months) were switched to a raltegravir-containing regimen. Blood samples were taken at baseline and at ≥2 timepoints up to 48±6 weeks. Levels of total HIV-1 DNA and 2-LTR circles in peripheral blood mononuclear cells (PBMCs) were measured using real-time PCR assays.

Results

There was no significant change in HIV-1 total DNA levels over the study duration (p = 0.808), median slope 0.24 (conservative nonparametric 95% CI: −11.78, 26.23). Low levels of 2-LTR circles were detected in 2 patients. One had 16 copies/10⁶ PBMCs at baseline and the other had 34 copies/10⁶ PBMCs at week 51.

Conclusions

The switch to a raltegravir containing regimen was not associated with a significant change in HIV-1 total DNA levels in this cohort. There were no observed changes in the levels of HIV-1 2-LTR circles associated with raltegravir treatment initiation.     

Cysteine peptidases, secreted by Trichomonas gallinae, are involved in the cytopathogenic effects on a permanent chicken liver cell culture

Trichomonas gallinae, the aetiological agent of avian trichomonosis, was shown to secrete soluble factors involved in cytopathogenic effect on a permanent chicken liver (LMH) cell culture. The present study focused on the characterization of these molecules. The addition of specific peptidase inhibitors to the cell-free filtrate partially inhibited the monolayer destruction, which implied the presence of peptidases in the filtrate and their involvement in the cytopathogenic effect. One-dimensional substrate (gelatin) SDS-PAGE confirmed the proteolytic character of the filtrate by demonstrating the proteolytic activity within the molecular weight range from 38 to 110 kDa. In addition, the proteolytic activity was specifically inhibited by addition of TLCK and E-64 cysteine peptidase inhibitors implying their cysteine peptidase nature. Furthermore, variations in the intensity and the number of proteolytic bands were observed between cell-free filtrates of low and high passages of the same T. gallinae clonal culture. Two-dimensional substrate gel electrophoresis of concentrated T. gallinae cell-free filtrate identified at least six proteolytic spots. The mass spectrometric analysis of spots from 2-D gels identified the presence of at least two different Clan CA, family C1, cathepsin L-like cysteine peptidases in the cell-free filtrate of T. gallinae. In parallel, a PCR approach using degenerated primers based on the conserved amino acid sequence region of cysteine peptidases from Trichomonas vaginalis identified the coding sequences for four different Clan CA, family C1, cathepsin L-like cysteine peptidases. Finally, this is the first report analyzing molecules secreted by T. gallinae and demonstrating the ubiquity of peptidases secreted by this protozoon.     

Characteristic new members of the phthiocerol and phenolphtiocerol families from Mycarbacterium ulcerans

Abstract Dtacyl phtluodlolone A and phenolphthtodlo-lone A hpids were isolated from two stratus of Mycobactertum ulcerans . The dlol umts of the phthione A and phenolphth]odlolone A components were shown to have erythro stercochemistry by infrared spectroscopy and proton nuclear magnetic resonance of an acetal derivative This stereoehemistry is shared only by related dvols from M. marmum , the dhols from M. boys, M kansasu, M leprae and M. luberculosiss having threo stereochemlstry.     

Effect of gamma irradiation on rheological properties of polysaccharides exuded by A. fluccosus and A. gossypinus

In this study, Iranian gum tragacanth (GT) exudates from Astragalus fluccosus (AFG) and Astragalus gossypinus (AGG) were irradiated at 3, 7, 10 and 15 kGy. Fourier transform infrared spectroscopy (FTIR) data showed that irradiation did not induce changes in the chemical structure of either type of gum. Although particle size distribution and both steady shear and dynamic rheological properties were considerably affected by the irradiation process, the magnitude of the effect of irradiation on each of the rheological and size variables was different for the hydrocolloids. For instance, for AGG, increasing the irradiation dose from 3 to 10 kGy, the d(0.5) and D[3,2] values were reduced by one-sixth to one-eighth fold. Colour measurement revealed that the radiation process led to an increase in the yellow index and b* values for both types of GT in powder form, but it was more pronounced for AGG samples. Irradiation led to an approximate 13-fold increase in redness in AFG. Surface and shape changes of the gum crystals were studied by scanning electron microscope (SEM) and a smoother surface for irradiated samples was detected. The notable changes in functional properties of each variety of irradiated gum should be taken into consideration before using the radiation technology as a commercial tool for sterilisation.