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The effect of geldanamycin (GA), a specific inhibitor of heat shock protein 90 (Hsp90), on gene expression and replication of human cytomegalovirus (HCMV) was studied in human embryonic lung (HEL) fibroblasts. Kinetic analysis indicated that GA delayed synthesis of major immediate early (MIE), early and late viral proteins, and blocked a second tier of the synthesis of these proteins that occurred in untreated cells after 48 h post-infection (pi). Moreover, when HCMV-infected HEL cells were maintained with medium containing 40 nM GA for 6 days, with medium changes at 2-day intervals, the virus yield was reduced to an undetectable level. On a molecular level, the cellular kinase Akt and the transcription factor NFkappaB were activated in HCMV-infected cells within 30 min pi. NFkappaB was shown to be essential for MIE gene expression. However, in GA-treated cells, activation of both Akt and NFkappaB was greatly inhibited. Because LY294002, an inhibitor of cellular phosphatidylinositol 3-kinase (PI3-K), also prohibited HCMV-mediated activation of Akt and NFkappaB and synthesis of the MIE proteins, PI3-K signalling was necessary for expressing the MIE genes. These results suggest that the inhibitory effect of GA on HCMV replication is primarily caused by the disruption of the PI3-K signalling pathway, leading to the activation of NFkappaB, which plays a crucial role in expression of the critical MIE genes.

As part of an ongoing cohort study in the Hokuriku region of Japan, cervical cell samples from histologically confirmed normal (n ‫؍‬ 114) or abnormal (n ‫؍‬ 286) women were examined for the presence of HPV DNA using a second-generation hybrid capture assay (HCA-II) and LCR-E7 PCR. HCA-II detected low-risk (HPV-6, -11, -42, 43 and -44) and high-risk (HPV-16, -18, -31, -33, -35, -39, -45, -51, -52, -56, -58, -59 and -68) HPV types, while LCR-E7 PCR detected an additional 7 HPV types and some uncharacterized types. In screening of high-grade squamous intraepithelial lesions (HSILs) and invasive cervical cancer, the sensitivities of HCA-II and LCR-E7 PCR testing the high-risk HPV types were 83% and 81%, respectively, while the specificity of both assays was 93%. The sensitivity of LCR-E7 PCR increased to 87%, which was significantly higher than that in HCA-II, when testing both highrisk and other HPV types. Sixty-eight inconsistent results (17% of total tested) from HCA-II and LCR-E7 PCR were due to (i) low copy number of HPV genome (false-negative for HCA-II, 5.3% and for LCR-E7 PCR, 1.3%), Key words: hybrid capture assay II; LCR-E7 PCR; human papillomavirus; cervical cancer; high-grade squamous intraepithelial lesionAlthough deaths from cervical cancer in many developed countries have declined in recent decades, it remains the fifth most frequent cancer and the second most common cancer in women worldwide. 1 The adoption of routine cytologic testing for cervical cancer in many developed countries accounts, in large part, for the decrease in deaths from this disease. Problems remain with cytologic testing, however, particularly with the frequency of falsenegatives, the high cost of repeat testing and diagnosis of equivocal cases using the Bethesda system. 2 Given these circumstances, cancer-screening programs using HPV tests are as effective at predicting disease as those using cytologic tests. 3 zur Hausen et al. 4 showed that infection with HPV was closely associated with cervical cancer development. In addition, several previous studies have shown that HPV-6 and -11 are associated with benign anogenital lesions, whereas HPV-16 and -18 are associated with cervical cancer. 5 Currently, more than 80 HPV types have been identified and of these, about 30 distinct HPV types are known to infect the genital tract, 6,7 at least 10 being associated with cancer. 8 Geographic differences in HPV types have been reported to exist between countries 8 and even within the United States. 9,10 The risk factors for cervical cancer and the prevalent HPV types in Japan 11,12 differ from those reported in Western countries. 8 -10 HPV-51, -52 and -58 are more prevalent, whereas HPV-18 and -45 are less prevalent in Japan than in Western countries. 12 Therefore, the definition of high-risk HPV types in Japanese women is important with respect to both clinical management of HPV infection and unequivocal diagnosis of cervical pathology.Highly sensitive HPV DNA tests have been developed as a supplements to cervical cancer screening and fo...

Based on knowledge from traditional Arab herbal medicine, this in vitro study aims to examine the anti-inflammatory mechanism of Hypericum triquetrifolium by measuring the expression and release of pro-inflammatory cytokines, tumor necrosis factor-α (TNF-α) and interleukine-6 (IL-6), and inducible nitric oxide synthase (iNOS) in human monocytic cells, THP-1. The effects were assessed by measuring the levels of secretory proteins and mRNA of TNF-α and IL-6, the levels of nitric oxide (NO) secretion and the expression of iNOS in THP-1 cells. Cells were treated with 5 μg lipopolysaccharide/ml (LPS) in the presence and absence of increasing concentrations of extracts from the aerial parts of H. triquetrifolium. During the entire experimental period, we used extract concentrations (up to 250 μg mL−1) that had no cytotoxic effects, as measured with MTT and LDH assays. Hypericum triquetrifolium extracts remarkably suppressed the LPS-induced NO release, significantly attenuated the LPS-induced transcription of iNOS and inhibited in a dose-dependent manner the expression and release of TNF-α. No significant effects were observed on the release of IL-6. Taken together, these results suggest that H. triquetrifolium probably exerts anti-inflammatory effects through the suppression of TNF-α and iNOS expressions.

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