Search

Since most patients with hepatitis C virus (HCV) infection now receive treatment irrespective of liver disease severity, special attention to patient quality of life (QoL), including psycho-social aspects, is required. No QoL questionnaire is specific for patients with HCV. To develop and validate a short Italian questionnaire (HepaDisk) assessing the QoL of patients affected by HCV with intuitive graphic results that is understandable by patients and physicians. A questionnaire, drafted by a steering committee, underwent a Delphi survey. A multicenter, observational study was conducted to validate the developed HepaDisk versus other tools (CLDQ-I, SF-36, WPAI:HCV), and to evaluate its correlation with disease severity in Italian patients with HCV. The 10-item questionnaire was validated in 214 patients. HepaDisk showed a high correlation with CLDQ overall score and WPAI:HCV activity impairment (Spearman's rank correlation: 0.651 and 0.595, respectively) and a lower correlation with SF-36. Strong internal consistency (Cronbach coefficient: 0.912), good test–retest reliability (Pearson's correlation coefficient: 0.789; 95% CI, 0.714–0.865), and responsiveness to changes among improved patients were demonstrated. HepaDisk is a reliable and user-friendly tool that can monitor disease impact on patient QoL over time, providing a visual representation easily understandable by both patients and physicians. [ABSTRACT FROM AUTHOR]

Background The used first generation protease inhibitors may be hampered by virological failure in partially interferon-sensitive patients. Aim To investigate early hepatitis C virus (HCV)-RNA decay and quasispecies modifications, and disclose viral dynamics underlying failure. Methods Viraemia decay at early time-points during telaprevir treatment was modelled according to Neumann et al. (1998). NS3-sequences were obtained by population-sequencing and ultradeep-454-pyrosequencing. Results 13 treatment-experienced (8 non-responders, 5 relapsers), and two cirrhotic naïve patients, received telaprevir + pegylated-interferon-α + ribavirin. Viraemia decay was biphasic. In all patients, first-phase was rapid and consistent, with a median [interquartile-range] viraemia decay of 2.8 [2.6–3.2] log IU/ml within 48 h. Second-phase decay was slower, especially in failing patients: 3/3 showed <1 log IU/ml decay between 48 h and 2 weeks, and HCV-RNA >100 IU/ml at week 2. Only one patient experiencing sustained viral response showed similar kinetics. By pyrosequencing, mutational freeze was observed in all 15 patients within the first 24 h, but only in patients with sustained response afterwards. Indeed, 2/2 failing patients showed early resistance, as minor (V36A-T54A: prevalence <26% at 48 h) or major (V36M/A-R155K: prevalence, 99.8% at week 2) variants. Conclusions Following telaprevir administration, first-phase HCV-RNA decay is consistent with mutational freeze and limited/no viral replication, while second-phase is significantly slower in failing patients (with appearance of resistance), suggesting the usefulness of early HCV-RNA monitoring. [ABSTRACT FROM AUTHOR]

Abstract: Background: HIV infection is an independent risk factor for cardiovascular disease. HIV-sustained impairment of endothelial progenitor cells (EPCs) could contribute to this process, so that it is important to assess whether antiviral therapy (ART) is able to revert these abnormalities. Methods: We quantified in 21 naïves and 34 treated patients two functionally distinct clonogenic progenitors which have been acknowledged important for vascular repair: the hematopoietic progenitor colony forming unit - endothelial cells (CFU-EC) and the true endothelial progenitor, the endothelial colony forming cells (ECFC). We correlated results obtained with conventional vascular risk factors and with HIV-related parameters. Results: We found that these progenitors behaved differently in naive and treated patients. In particular, CFU-EC level was significantly low in all naive patients and slowly recovered during ART. In contrast, the ECFC level was abnormally high in naive patients while it decreased upon ART. The CFU-EC level was related to conventional cardiovascular risk factors, as reported in general population, but also to inflammatory indexes and CD4 cell count. In contrast, the ECFC number was exclusively related to viral replication activity and to CD4 cell count. Conclusions: In HIV-infected people, the levels of CFU-EC and ECFC are related to classical cardiovascular risk factors but, in addition, they are also significantly influenced by the infection itself and by antiviral therapy. [Copyright &y& Elsevier]

Chemokines are cytokines with chemotactic properties on inflammatory cells and other cell types. Chemokine (C-C motif) ligand 2 (CCL2), which is also called monocyte chemotactic protein 1 (MCP-1), is a potent chemotactic molecule that attracts lymphocytes, monocytes, mast cells, and memory T cells, but not neutrophils. CCL2/MCP-1 represents a link between the activation of monocytes, lymphocytes, basophils, mast cells, and eosinophils in inflammatory disorders, such as the late-phase allergic reaction. This C-C chemokine also plays a role in regulating Th-cell cytokine production and leukocyte trafficking. Laboratory of allergic diseases (LAD) cells is the first reported human mast cell line that closely resembles a primary culture of CD34+-derived human mast cells. These cells were cultured in vitro and treated with different concentrations of substance P (SP) for the production of CCL2/MCP-1. We used calcium ionophore as a positive control for stimulating transcription and translation of CCL2/MCP-1. The stimulation of SP on CCL2/MCP-1 was statistically significant (P <0.05) compared with the control (untreated cells). In this study, we determined the expression and secretion of CCL2/MCP-1 from SP-activated LAD2 human mast cells in vitro. The levels of CCL2/MCP-1 from SP-activated LAD2 human mast cells were higher at 10 μM and at 18 h incubation compared with controls. This effect was also revealed on CCL2/MCP-1 messenger RNA (mRNA) expression, as determined by reverse transcriptase polymerase chain reaction (RT-PCR) analysis. Our data suggest that SP is an important neurotransmitter that can stimulate the chemokine CCL2, which plays a fundamental role in inflammation by recruiting inflammatory cells to specific cites. [Copyright &y& Elsevier]

Background: Few studies are available on possible hepatitis C virus (HCV)-specific T- cell immune response in health care workers (HCWs) involved in the care of patients with HCV infection. We aimed to investigate whether a HCV-specific interferon (IFN)-¿ T-cell response, known to be involved in infection resolution, was present in those HCWs involved in the management of patients with persistent HCV infection. Methods: Our study involved 30 subjects, classified as group A (20 consecutive patients, 16 males and 4 females, with histologically proven chronic hepatitis), or group B (10 HCWs, 7 males and 3 females, with at least 7 years of health care experience and HCV-RNA and anti-HCV negative). As a control group, we used 10 blood samples from healthy donors at a blood donor center (group C). HCV-RNA was measured by real-time polymerase chain reaction. Blood samples (at least 35 mL) were collected from all group A and group B subjects in our hospital. Specific IFN-¿ was stimulated with HCV pool peptides (core, 2 ¿g/mL), with influenza Mp peptides used as a positive control. Results: Levels of HCV-specific IFN-¿¿positive cells were higher in the HCWs (group B) compared with the infected patients (group A) and healthy blood donors (group C) (Mann-Whitney U test, P < .001). Conclusion: A clinically silent persistent exposure to HCV, through some as-yet undetermined mechanism, may induce a virus-specific IFN-¿¿producing CD8+ T-cell response in healthy aviremic HCWs. This finding suggests that possible unapparent parenteral routes may stimulate host defenses with no evidence of hepatitis. [Copyright &y& Elsevier]

Abstract: To obtain information on the mechanisms responsible of the generation of ragged red fibers (RRF) during aging, we analyzed the mitochondrial genotype of single skeletal muscle fibers of healthy individuals having an age comprised between 45 and 92 years. The sequencing of the D-loop region showed many sequence changes with respect to the Cambridge reference sequence (CRS), in both RRF and normal fibers. These changes were more abundant in RRF and their number increased between 50 and 60, and 61 and 70 years and then remained approximately constant. The analysis of the sequence changes showed that each subject contained one or more changes associated to RRF in positions of D-loop region that either do not change or that change very rarely. In general the same type of RRF-associated change was not found in more than one individual; exceptions were changes in positions 189, 295, 374 and 514, detected in 20–50% of analyzed subjects. In particular the A189G age-associated mutation was found only in old individuals and prevalently in RRF. Sequencing of other two mtDNA regions showed no relevant changes in the 16S/ND1 region and two RRF-associated original mutations, G5847A and A5884C, in two very conserved positions of tRNATyr. These results indicate that each subject has its own pattern of RRF-associated mutations in both coding and non-coding region of human mtDNA. [Copyright &y& Elsevier]

Type II fiber loss and reactive oxygen species (ROS)-induced damage are hallmarks of muscle aging. The aim of this study was to analyze whether there exists a relationship between age-dependent changes in cellular antioxidant capacity and type II fiber loss in aged human skeletal muscles. Forty-five male and female subjects ranging in age from 65 to 90 year-old were divided into +40 and −40% type II fiber groups. We measured both total and Mn superoxide dismutase (total and MnSOD), glutathione peroxidase (GSHPx) and catalase (CAT) activities. We also measured the reduced and oxidized forms of glutathione (GSH and GSSG) and lipid peroxide (LPO) levels. Total SOD activity was lower in the −40% type II fiber group than in the +40% group; MnSOD tended to be lower but data are not statistically consistent. Both GSHPx and CAT activities remained unchanged; as did GSH, GSSG and GSH/GSSG ratio. Finally, muscle samples with −40% type II fibers had a significantly higher LPO content compared to those with +40% type II fibers. In summary, a relationship between human skeletal muscle aging, type II fiber loss and ROS reactions seems to exist. [Copyright &y& Elsevier]

Abstract: The aim of the study was to investigate the efficacy of the quorum sensing inhibitor FS3 and daptomycin in preventing prosthesis biofilm in a rat model of staphylococcal vascular graft infection. Graft infections were established in the back subcutaneous tissue of adult male Wistar rats by implantation of Dacron prostheses followed by topical inoculation with 2×107 colony-forming units of Staphylococcus aureus, strain Smith diffuse. The study included a control group, a contaminated group that did not receive any antibiotic prophylaxis and three contaminated groups that received: (i) intraperitoneal daptomycin, (ii) FS3-soacked graft, and (iii) daptomycin plus FS3-soaked graft, respectively. Each group included 15 animals. The infection burden was evaluated by using sonication and quantitative agar culture. Moreover, an in vitro binding-study was performed to quantify the how much FS3 was coated to the surface of the prosthesis. The in vitro studies showed, that minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values for daptomycin were lower in presence of FS3. In in vivo studies, when tested alone, daptomycin and FS3 showed good efficacies. Their combination showed efficacies significantly higher than that of each single compound. Daptomycin is an important candidate for prevention of staphylococcal biofilm related infection and FS3 could serve as an interesting anti-staphylococcal antibiotic enhancer. [Copyright &y& Elsevier]

Background: Alkaline phosphatase (ALPase) is found in blood plasma or serum and leukocytes and regulates intercellular processes, maintaining phosphoryl metabolites in a steady state, as well as synthesizing and hydrolyzing phosphate esters on membranes. ALPase supervises the active transport of inorganic phosphates, fats, proteins, carbohydrates and the sodium/potassium pump mechanisms. The formed elements of blood such as polymorphonuclear (PMNs) leucocytes, macrophages (MP) and some lymphocytes are high in ALPase concentrations. Methods: In this study we have tested whether the interleukin-1 receptor antagonist (IL-lra) could influence ALPase generation in IL-1β or lipopolysaccharide (LPS)-stimulated neutrophils and MP. Human neutrophils were isolated from heparin-anticoagulated blood drawn from healthy individuals by centrifugation in a two-step gradient, Ficoll-Hypaque. ALPase activity was assessed spectrophotometrically in test tubes containing isolated neutrophils and adherence PBMCs treated with LPS, IL-1β and IL-1ra, alone or in combination. Results: IL-lβ or LPS enhanced ALPase in both PMNs and MP, whereas IL-1ra could not inhibit ALPase activity. We performed time course experiments at 0 min, 5 min, 1 h, 24 h, and 43 h (LPS 20 μg/mL, IL-1β 10 ng/mL). No significant increase in ALPase activity was seen until 1 h; however, there was a rapid rise over the next few hours. In another set of experiments using IL-1ra (500 ng/mL), there was no difference between treated cells and control cells. The combination of IL-1β plus IL-1ra did not reduce the ability of IL-1β to induce ALPase activity. Conclusions: These data suggest that IL-1β stimulates ALPase through other mechanisms than the release of arachidonic acid products, which are inhibited by IL-lra. [Copyright &y& Elsevier]

Abstract: Aging is related to the accumulation of reactive oxygen species (ROS)-mediated oxidative damage. Considering the heterogeneity of age-related changes and the involvement of muscles in different functions, we compared the aging process in different functional muscles. We studied age-related changes in rectus abdominis (RA) and vastus lateralis (VL) in subjects of different age (18–48- and 66–90-year-old). We analysed fiber distribution, antioxidant enzymatic systems: Mn and CuZn superoxide dismutase (MnSOD, CuZnSOD), glutathione peroxidase (GSHPx), catalase (CAT), as well as oxidative damage markers: lipoperoxide levels (LPO), carbonylated proteins (CP), reduced and oxidized glutathione (GSH, GSSG) content and the GSH/GSSG ratio. In the muscles analysed, type I fiber increases during aging with a consequent decrease in type II distribution. In the elderly group RA MnSOD showed higher activity than VL. Furthermore, in RA MnSOD was higher in the elder group than in the younger group. CuZnSOD, as well as GSHPx and CAT activities remained unchanged. LPO levels in VL increase with age; moreover, in the elderly group VL showed higher value than RA. CP, GSH and GSSG remained unchanged, while GSH/GSSG decreases in RA during aging. In conclusion, a relationship between aging and ROS seems to exist, but oxidative processes could evolve in different ways in muscles with different functions. [Copyright &y& Elsevier]