A recombinant adenovirus encoding hepatitis C virus core and E1 proteins protects mice against cytokine-induced liver damage

Hepatitis C virus (HCV) infection has a strong tendency to evolve to chronicity despite up-regulation of proapoptotic cytokines in the inflamed liver. The mechanisms responsible for persistent viral replication in this inflammatory environment are obscure. It is conceivable that viral replication would be facilitated if the infected hepatocytes are rendered resistant to cytokine-induced cytotoxicity. In this study, we investigated if an adenovirus encoding HCV core and E1 (RAdCE1) could reduce liver cell injury in different in vivo models of cytokine-mediated hepatotoxicity in mice. We show that RAdCE1 markedly attenuates hepatocellular apoptosis and the increase in serum transaminase levels after concanavalin A (con A) challenge. This protective effect is accompanied by an inhibition of nuclear translocation of nuclear factor kappaB (NF-kappaB); reduced expression of inducible nitric oxide synthase (iNOS); decreased hepatic messenger RNA levels of chemokines macrophage inflammatory protein 2 (MIP-2), monocyte chemoattractant protein 1 (MCP-1), and interferon-inducible protein 10 (IP-10); and abrogation of liver leukocyte infiltration. RAdCE1 also causes a reduction in serum transaminase levels and inhibits hepatocellular apoptosis in mice given tumor necrosis factor (TNF)-alpha plus D-galactosamine. In conclusion, HCV structural antigens can protect liver cells against the proapoptotic effects of proinflammatory cytokines. The antiapoptotic status of infected liver cells may represent a mechanism favoring viral persistence. Our findings also suggest that, in chronic hepatitis C, the burden of hepatocellular damage mainly affects noninfected liver cells.     

MicroRNAs as potential regulators of platelet function and bleeding diatheses

Although a growing number of studies suggest that microRNAs (miRNAs) play a relevant role in platelet biology, their implications in bleeding diatheses are starting to be investigated. Indeed, several studies have shown that alterations in the intracellular levels of highly expressed platelet miRNAs provoke a thrombotic phenotype. On the other hand, primary immune thrombocytopenia (ITP), which is considered the hallmark of acquired bleeding disorders, has been recently associated with altered levels of miRNAs in peripheral blood mononuclear cells, plasma, and platelets. In this review, we will focus on miRNAs that may affect the hemostatic and thrombotic functions of platelets, and we will discuss the different studies that have attempted to associate miRNAs with regulatory mechanisms of ITP.     

Determination of Clara cell protein urinary elimination as a marker of tubular dysfunction

Clara cell 16-kDa protein (CC16) is a protein expressed primarily by the bronchial cells. It is rapidly eliminated by glomerular filtration, reabsorbed almost entirely, and catabolized in proximal tubule cells. To date, normal values for urinary CC16 in healthy children have not been determined. We have studied 63 pediatric patients (mean age 8.17 ± 3.91 years) and 31 healthy children (control group; mean age 8.83 ± 3.65 years). In the control group, the CC16/creatinine ratio was 1.22 ± 1.52 μg/g. In 16 out of 31 control children, the value of the ratio was zero. Fourteen patients (22.2%) showed a high CC16/creatinine ratio; in contrast, among these same patients, the ratio N-acetyl-β-d-glucosaminidase (NAG)/creatinine was elevated in seven cases (11.1%) and the ratio β2-microglobulin/creatinine was elevated in seven cases (11.1%). The three parameters were in agreement in 51 patients (80.9%). Among the patients, the CC16/creatinine ratio was correlated with both the β2-microglobulin/creatinina ratio (r = 0.76, P < 0.001) and the NAG/creatinine ratio (r = 0.6, P < 0.001). Our findings indicate that CC16 is a good marker of proximal tubular function in childhood. The highest observed values were in children with proximal tubulopathies, in children with chronic renal failure, and in those treated with cyclosporine.     

Effect of surface treatments on the bond strength of brackets bonded with a compomer

PURPOSE: To evaluate the effectiveness of the compomer Dyract AP for bonding brackets when conditioning the enamel with phosphoric acid and a non-rinsing conditioner (NRC), in comparison with a control group in which the resin orthodontic adhesive system Transbond XT was used. METHODS: The brackets were bonded to extracted premolars which were divided into three groups: (1) Acid/Transbond XT, (2) NRC/Dyract AP and (3) Acid/Dyract AP. Shear bond strength was measured with a universal testing machine. The crosshead speed was 1mm/minute. The adhesive remnant on the tooth after debonding was determined using image analysis equipment. RESULTS: The bond strength of Acid/Transbond XT was significantly higher than bond strengths of Acid/Dyract AP and NRC/Dyract AP (P< 0.017). No significant differences were observed between Acid/Dyract AP and NRC/Dyract APBond strength values (P> 0.017). Acid/Transbond XT left significantly more adhesive on the tooth than Acid/Dyract AP and NRC/Dyract AP, whereas NRC/Dyract AP left significantly less adhesive than Acid/Dyract AP (P< 0.05).     

Influence of fluoride varnish on shear bond strength of a universal adhesive on intact and demineralized enamel

The aim was to evaluate the effect of fluoride varnish on the shear bond strength (SBS) on polished and non-polished intact and demineralized enamel. Bovine incisors (half demineralized) were used. Bifluorid 12? was applied. Bonding was made with Futurabond^®M?+?and GrandioSO, 24 h and 7 days after varnishing. In some groups, varnish was removed by polishing before bonding. SBS was measured. Fracture type was determined by stereomicroscopy and scanning electron microscope (SEM) observations of the enamel surface were made. Between-group differences were determined by one-way ANOVA and the Tukey test. Associations between study factors and fracture modes were analysed using contingency tables and Pearson’s chi-squared test. For intact enamel, SBS on varnished enamel at 24 h was significantly less than in the other groups. SBS recovered 7 days after varnishing. Varnish elimination after 24 h significantly increased the SBS. However, removal at 7 days did not modify SBS. SBS on demineralized enamel groups was significantly less than in intact enamel, except for demineralized enamel varnished and removed at 7 days. Demineralized enamel was associated with cohesive enamel fractures and intact enamel with cohesive fractures of the composite and adhesive fractures. SEM of varnish surfaces showed a homogenous layer scattered with amorphous precipitate. In conclusion, on intact enamel fluoride varnish had a negative effect on SBS at 24 h, which disappeared after 7 days. On demineralized enamel, varnish did not reduce SBS at either time. Polishing the varnished enamel surface showed a similar SBS to intact enamel after 7 days.     

A multicenter phase II study of irinotecan in patients with advanced colorectal cancer previously treated with 5-fluorouracil

This multicenter, open-label, phase II study was performed to assess the efficacy and toxicity of irinotecan 350 mg/m2 intravenously every 3 weeks in patients with advanced colorectal cancer (CRC) previously treated with 5-fluorouracil (5-FU). The study enrolled 115 patients and a total of 558 cycles (median, 6 per patient) were administered. The overall objective response rate on an intent-to-treat basis was 18% (with 1 complete response and 20 partial responses), whereas 42 patients (37%) showed stable disease. Median time to progression was 4.8 months and median survival was 13.6 months. Grade 3/4 toxicities included delayed diarrhea (19.1%), nausea/vomiting (10.4%), and neutropenia (8.7%). There were 2 toxic deaths, 1 from delayed diarrhea and 1 from hemorrhage and grade 4 mucositis. In conclusion, the present study confirms the antitumor efficacy of irinotecan monotherapy in patients with CRC pretreated with 5-FU.     

Vaccination with an adenoviral vector encoding hepatitis C virus (HCV) NS3 protein protects against infection with HCV-recombinant vaccinia virus

Cellular immune response plays an important role in the clearance of hepatitis C virus (HCV). Thus, development of efficient ways to induce anti-viral cellular immune responses is an important step toward prevention and/or treatment of HCV infection. With this aim, we have constructed a replication-deficient recombinant adenovirus expressing HCV NS3 protein (RAdNS3). The efficacy of RAdNS3 was tested in vivo by measuring the protection against infection with a recombinant vaccinia virus expressing HCV-polyprotein (vHCV1-3011). Immunisation with 10(9)pfu of RAdNS3 induced anti-NS3 humoral, T helper and T cytotoxic responses. We identified eight epitopes recognised by IFN-gamma producing cells, five of them exhibiting lytic activity. Moreover, we show that RAdNS3 immunised mice were protected against challenge with vHCV1-3011 and that this protection was mediated by CD8(+) cells. In conclusion, our results suggest that adenoviral vectors encoding NS3 might be useful for the induction of prophylactic and/or therapeutic anti-HCV immunity.