HCV Induces Oxidative and ER Stress, and Sensitizes Infected Cells to Apoptosis in SCID/Alb-uPA Mice

Hepatitis C virus (HCV) is a blood-borne pathogen and a major cause of liver disease worldwide. Gene expression profiling was used to characterize the transcriptional response to HCV H77c infection. Evidence is presented for activation of innate antiviral signaling pathways as well as induction of lipid metabolism genes, which may contribute to oxidative stress. We also found that infection of chimeric SCID/Alb-uPA mice by HCV led to signs of hepatocyte damage and apoptosis, which in patients plays a role in activation of stellate cells, recruitment of macrophages, and the subsequent development of fibrosis. Infection of chimeric mice with HCV H77c also led an inflammatory response characterized by infiltration of monocytes and macrophages. There was increased apoptosis in HCV-infected human hepatocytes in H77c-infected mice but not in mice inoculated with a replication incompetent H77c mutant. Moreover, TUNEL reactivity was restricted to HCV-infected hepatocytes, but an increase in FAS expression was not. To gain insight into the factors contributing specific apoptosis of HCV infected cells, immunohistological and confocal microscopy using antibodies for key apoptotic mediators was done. We found that the ER chaperone BiP/GRP78 was increased in HCV-infected cells as was activated BAX, but the activator of ER stress–mediated apoptosis CHOP was not. We found that overall levels of NF-κB and BCL-xL were increased by infection; however, within an infected liver, comparison of infected cells to uninfected cells indicated both NF-κB and BCL-xL were decreased in HCV-infected cells. We conclude that HCV contributes to hepatocyte damage and apoptosis by inducing stress and pro-apoptotic BAX while preventing the induction of anti-apoptotic NF-κB and BCL-xL, thus sensitizing hepatocytes to apoptosis

Estimation of relative fecundity in<i>Eimeria tenella</i>strains by a mixed infection method

SUMMARYThe relative fecundity of populations ofEimeria tenellawas estimated by means of mixed infection using electrophoretic variation of glucose-phosphate isomerases (GPIs) as a genetic marker. A decoquinate-resistant strain with GPI-9 isozyme (DR-NIAH), a decoquinate-sensitive one with GPI-1 (DS-Iwate), and three decoquinate-resistant lines (No. 3, 4, and 5) derived from cross-fertilization between DR-NIAH and DS-Iwate, were used. The GPI phenotypes of the No. 3 and No. 4 lines are GPI-9, and that of No. 5 is GPI-1. Mixed infection experiments were performed between DR-NIAH and DS-Iwate, No. 3 and No. 5, and No. 4 and No. 5. DR-NIAH was predominant over DS-Iwate in the mixed infection, whereas, in single infections, the total oocyst output of DR-NIAH was similar to or less than that of DS-Iwate. Among three lines, No. 4 was predominant over No. 5, and No. 5 was predominant over No. 3 in the mixed infection. Relative fecundity between No. 3 and No. 5 and their patterns of oocyst output in single infections were similar to those in the mixed infection. In contrast, No. 5 produced more oocysts than No. 4 in single infections, suggesting that the oocyst production in the mixed infection may be influenced by the mutual interference or competition between the populations ofE. tenellain the chicken caeca.</jats:p

Effect of Tween 80 on formation of the superficial L1 layer of the Mycobacterium avium-Mycobacterium intracellulare complex

The effects of Tween 80 supplementation of liquid culture medium on the formation of the superficial L1 layer of the Mycobacterium avium-Mycobacterium intracellulare complex (MAC) were examined by serological and scanning electron microscopic experiments. Specific antiserum to the glycopeptidolipids on the L1 layer of M. avium S-139, made in a rabbit, was used for seroagglutination reactions with antigens prepared from strain S-139 grown in medium supplemented with various levels of Tween 80 (0, 0.05, 0.5, 5, and 50 mg/ml). The agglutination titers gradually decreased as the concentration of Tween 80 rose. Scanning electron microscopy showed that the fibrillar materials consisting mainly of glycopeptidolipids on the L1 layer of strain S-139 also disappeared with increases in the concentration of Tween 80. In addition, there was no obvious correlation between (i) the plasmid DNAs and serotypes of MAC and (ii) formation of the L1 layer of MAC. It is therefore concluded that Tween 80 used to supplement liquid culture medium affects formation of the L1 layer, which has been considered to be one of the pathogenic factors of MAC.</jats:p