Hepatitis C virus E2 envelope protein induces dendritic cell maturation.

Maturation is a critical process for dendritic cells (DC) to gain or enhance their functions in antigen presentation and T-cell activation. In this study, we investigated the effect of hepatitis C virus (HCV) envelope protein E2 on DC maturation and related functions. We show that binding of E2 protein to DC leads to a change from immature to mature phenotype as detected by an increased expression of cell surface molecules including CD83, CD80, CD86, CD11c and MHC class II. The E2-matured DC showed higher capacity to stimulate T-cell proliferation and interferon-gamma production and displayed higher levels of interleukin-12 production when compared with immature DC. The induction of DC maturation by E2 is both time- and dose-dependent and can be inhibited by anti-E2 antibodies. In addition, DC matured by E2 showed decreased uptake of bovine serum albumin and latex beads, indicating their decreased activities of endocytosis and phagocytosis upon maturation. Taken together, our results demonstrated that E2 protein is able to induce dendritic cell maturation and suggested that E2 protein may play an important role in regulation of immune responses during HCV infection.

Varicose veins possess greater quantities of MMP-1 than normal veins and demonstrate regional variation in MMP-1 and MMP-13.

BACKGROUND: Studies have reported that structural proteins such as elastin and collagen are decreased in varicose veins compared to normal controls. We hypothesized that the changes observed in varicose vein wall composition may be related to alterations in extracellular matrix remodeling proteins, such as the matrix metalloproteases and serine proteases. In addition we hypothesized that there may be regional variation in the expression of these enzymes within the leg. PATIENTS AND MATERIALS: One-centimeter segments of the proximal and distal greater saphenous vein (GSV) were obtained from patients undergoing ligation and stripping for venous insufficiency (vv) (n = 15) or GSV harvest in conjunction with coronary artery bypass grafting (CABG) (n = 7). All vv patients had incompetence of the GSV by color flow duplex. Vein specimens were examined for MMP-1, 3, and 13, tryptase, and GAPDH mRNA using semiquantitative RT-PCR analysis. Quantification of MMP-1 and 13 (active/latent forms) and tryptase was performed using Western blot analysis. Western blots were analyzed using scanning densitometry and standardized to normal controls and values expressed as the median densitometric index (D.I.). Nonparametric statistical methods (Wilcoxan signed rank test and Mann-Whitney U test) were used for analysis. RESULTS: We were able to amplify MMP-1, MMP-13, and tryptase mRNA from both proximal and distal segments of all greater saphenous veins studied. MMP-3 mRNA, however, was not found in either segment of any of the veins examined. A semiquantitative analysis of RT-PCR products comparing the ratio of MMP-1, MMP-13, or tryptase mRNA to GAPDH mRNA showed no difference between cases and controls nor proximal vs distal vein segments. Western blot analysis revealed larger quantities of MMP-1 in varicose veins than in nondiseased veins from CABG patients (48.0 +/- 36.7 D.I. vs 12.5 +/- 6.8 D.I., P = 0.036). Investigation into the regional variation of proteases revealed lower amounts of MMP-1 in distal than in proximal vein segments (37.9 +/- 35.0 D.I. vs 44.1 +/- 41.6 D.I., P = 0.01). Similarly, we found significantly less MMP-13 in distal segments of varicose veins than in proximal segments (152.8 +/- 130.0 D.I. vs 206.7 +/- 173.3 D.I., P = 0.006). CONCLUSIONS: This study found that MMP-1 protein is increased in varicose veins when compared to controls despite no differences in mRNA expression. In addition we found that there is regional variation of MMP-1 and MMP-13 in diseased varicose veins. Lower leg veins have significantly reduced amounts of these proteolytic enzymes when compared to veins of the upper thigh. These data suggest that posttranscriptional regulatory controls could be responsible for the observed differences.

Stability of cisplatin, doxorubicin, and mitomycin combined with loversol for chemoembolization.

OBJECTIVE: To evaluate the chemical stability of a mixture of cisplatin 10 mg/mL, doxorubicin 5 mg/mL, and mitomycin 1 mg/mL in sodium chloride 0.9% (NS), and sodium chloride 0.9% and ioversol 68% (NSI). METHODS: The agents were reconstituted with NS and NSI in two groups. One syringe was made and stored protected from light at 4, 25, and 37 degrees C. Triplicate HPLC determinations were performed on each syringe at each time period. Drug concentration at T0, 4, 12, 24, 72, and 120 hours were recorded. RESULTS: Cisplatin in NS remained stable for five days at all three temperatures and lost 12.7% at 37 degrees C in 120 hours in NSI. Doxorubicin in NS remained stable throughout the five days at 4 and 25 degrees C, but lost 13.1% at 37 degrees C in 24 hours. Doxorubicin in NSI remained stable for five days at 4 degrees C and lost 16.3% at 25 degrees C by day 5, and 18.8% at 37 degrees C in 24 hours. Mitomycin in NS lost 15.8% at 4 degrees C within 24 hours, and 24.7% at 25 degrees C and 36% at 37 degrees C within four hours. Mitomycin in NSI was stable for 72 hours at 4 degrees C, 24 hours at 25 degrees C, and less than four hours at 37 degrees C. CONCLUSIONS: These agents in NS are stable for 12 hours at 4 degrees C. In NSI, they are stable for 72 hours at 4 degrees C.