ABSTRACT
Multidrug resistance associated with the overexpression of ATP-dependent binding cassette (ABC) proteins is widely accepted as an important cause of treatment failure in patients with neoplastic or infectious diseases. Some of them play also a pivotal role in detoxification processes. Herein, we investigated the effect of hepatitis C virus (HCV) replication and nonstructural 5A (NS5A) protein on the expression and functional activity of two ABC transport proteins: MDR1 and BCRP. RT-quantitative real-time polymerase chain reaction (qPCR) was carried out for mdr1 and bcrp mRNAs in both Huh7 cells expressing NS5A and Huh7.5 cells containing either full-length- or subgenomic-HCV replicon systems. The functional activity of these pumps was studied by performing a dye efflux assay with DiOC2 and Rhodamine 123. A dose-dependent down-regulation of mdr1 expression was documented in Huh7 cells expressing the NS5A protein, as well as in both replicon systems. In contrast, a significant increase of bcrp expression in both systems was recorded, which were in full agreement with the dye efflux assay results. These results warrant further in vivo studies in HCV patients with cholestasis and/or patients that are refractive to the pharmacotherapy due to the activity of these pumps.
Subject(s)
ATP Binding Cassette Transporter, Subfamily B, Member 1/biosynthesis , Gene Expression , Hepacivirus/physiology , Proto-Oncogene Proteins c-bcr/biosynthesis , Viral Nonstructural Proteins/metabolism , Virus Replication , ATP Binding Cassette Transporter, Subfamily B , Carbocyanines/metabolism , Cell Line , Gene Expression Profiling , Hepatocytes/virology , Humans , Real-Time Polymerase Chain Reaction , Rhodamine 123/metabolismABSTRACT
During two years (1997-1999) an investigation of possible infections of chlamydial etiology in outpatients with follicular conjunctivitis was carried out, through the use of specific assays. Fifty-seven selected patients with presumptive inclusion conjunctivitis were diagnosed by means of ophthalmoscopic examination and bilateral tarsal-conjunctiva swabbing for microorganisms. The possible presence of Chlamydia trachomatis was tested by immunofluorescence microscopy and isolation in cell culture of McCoy line. Of the 57 conjunctivitis patients screened, 37 (65%) proved to be positive by cell culture (CC) and 27 (47%) by direct immunofluorescence (IFD). A good agreement between the two assays was observed, where the CC was more sensitive than IFD. Of these 37 patients with chlamydial conjunctivitis, 23 (62%) were women, with over one-third of them ranging in age from 45 to 65 years. Their clinical records revealed an evolution period of 1 to 12 months. Eighteen (78%) of these women reported previous genital pathology, while 4 (29%) of the 14 men had a history of urethritis by Chlamydia trachomatis. A high frequency of follicular conjunctivitis by Chlamydia (65%) in the screened patients was observed, without any evidence of urogenital signs and symptoms at the moment of the study.
Subject(s)
Chlamydia trachomatis/isolation & purification , Conjunctivitis, Inclusion/microbiology , Adolescent , Adult , Aged , Cell Culture Techniques , Conjunctivitis, Inclusion/diagnosis , Female , Fluorescent Antibody Technique , Humans , Male , Middle AgedABSTRACT
During two years (1997-1999) an investigation of possible infections of chlamydial etiology in outpatients with follicular conjunctivitis was carried out, through the use of specific assays. Fifty-seven selected patients with presumptive inclusion conjunctivitis were diagnosed by means of ophthalmoscopic examination and bilateral tarsal-conjunctiva swabbing for microorganisms. The possible presence of Chlamydia trachomatis was tested by immunofluorescence microscopy and isolation in cell culture of McCoy line. Of the 57 conjunctivitis patients screened, 37 (65
) proved to be positive by cell culture (CC) and 27 (47
) by direct immunofluorescence (IFD). A good agreement between the two assays was observed, where the CC was more sensitive than IFD. Of these 37 patients with chlamydial conjunctivitis, 23 (62
) were women, with over one-third of them ranging in age from 45 to 65 years. Their clinical records revealed an evolution period of 1 to 12 months. Eighteen (78
) of these women reported previous genital pathology, while 4 (29
) of the 14 men had a history of urethritis by Chlamydia trachomatis. A high frequency of follicular conjunctivitis by Chlamydia (65
) in the screened patients was observed, without any evidence of urogenital signs and symptoms at the moment of the study.
ABSTRACT
In this 4-year follow-up in vivo controlled study, 112 human permanent first molars from children between 6 and 11 years old were used to investigate the viability of the carbon dioxide (CO2) laser in promoting caries-free occlusal surfaces in permanent molars as an isolated form of treatment or associated with conventional fissure sealants. The findings suggest that occlusal caries prevention only by means of CO2 laser irradiation is not effective; that the utilization of photoactivated sealants, as well as its association with CO2 laser, applied over the occlusal fissures, are effective means of preventing occlusal caries, and that the application of CO2 laser over occlusal fissures prior to the application of a photoactivated fissure sealant improves the retention of the sealant.