HPV infections are believed to be a necessary cause of cervical cancer. Viral burden, as a surrogate indicator for persistence, may help predict risk of subsequent SIL. We used results of HPV test and cytology data repeated every 4 -6 months in 2,081 women participating in a longitudinal study of the natural history of HPV infection and cervical neoplasia in São Paulo, Brazil. Using the MY09/11 PCR protocol, 473 women were positive for HPV DNA during the first 2 visits. We retested all positive specimens by a quantitative, low-stringency PCR method to measure viral burden in cervical cells. Mean viral loads and 95% CIs were calculated using log-transformed data. RRs and 95% CIs of incident SIL were calculated by proportional hazards models, adjusting for age and HPV oncogenicity. The risk of incident lesions increased with viral load at enrollment. The mean number of viral copies/cell at enrollment was 2.6 for women with no incident lesions and increased (trend p ‫؍‬ 0.003) to 15.1 for women developing 3 or more SIL events over 6 years of follow-up. Compared to those with <1 copy per cell in specimens tested during the first 2 visits, RRs for incident SIL increased from 1.9 (95% CI 0.8 -4.2) for those with 1-10 copies/cell to 4.5 (95% CI 1.9 -10.7) for those with >1,000 copies/cell. The equivalent RR of HSIL for >1,000 copies/cell was 2.6 (95% CI 0.5-13.2). Viral burden appears to have an independent effect on SIL incidence. Measurement of viral load, as a surrogate for HPV persistence, may identify women at risk of developing cervical cancer precursors. © 2002 Wiley-Liss, Inc. Key words: human papillomavirus; cervical neoplasia; cohort study; longitudinal analysis; viral loadEstablishment of productive infections by oncogenic types of HPV is a key early event in the natural history of cervical cancer. Given the positive relationship between viral load and the likelihood of persistent HPV infection 1 and the strong relationship between the latter and risk of cervical neoplasia, 2 a single measurement of viral load in cervical specimens may be a suitable biomarker of either a transient infection, or the likelihood that an instance of HPV DNA positivity may represent a persistent infection or one that may become persistent over time and lead to the development of cervical SIL. In the latter case, little is known, however, about the relationship between level of viral burden and lesion incidence or lesion grade severity in the natural history of CIN. HPV DNA viral load has been identified as a biomarker for cervical lesions in women, 3,4 primarily with minor-grade cytologic atypia. 5 However, only a few studies have evaluated the predictive effect of viral load on the incidence of cervical cancer precursors. 3,4,6 -8 A few methods of viral load estimation have been evaluated in epidemiologic studies. Although several cross-sectional studies have examined the association between HPV load measured by HC and the presence of SIL, 9 -13 HC cannot accurately quantify HPV DNA in cervical specimens because of the variabl...

Results confirm the gender-dependence of creatinine concentrations in spot specimens and also show age-dependence, indicating the need for these aspects to be considered when the range of acceptable samples is to be set. No significant intra- or inter-day variations were observed for the two parameters. Lastly, the possibility of a comparison of differently adjusted values was indicated by a conversion formula derived from adjustments to creatinine and the corresponding specific gravity of a hypothetical urinary value, as follows: specific gravity adjusted values = 1.48 x creatinine adjusted values.

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The presence of albumin receptors on the plasma membrane of isolated human hepatocytes was investigated employing albumin-coupled latex minibeads. Hepatocyte-latex reaction was visualized by phase contrast and scanning electron microscopy.The experiments demonstrate that hepatocytes exhibit binding activity for polymeric and monomeric forms of glutaraldehyde-treated albumin. Additionally, the reaction was shown to be speciesnonspecific.These findings support the hypothesis that polymerized albumin may act as a bridge between receptors on hepatitis B virus and human hepatocytes.Considerable data support a role of polymerized human serum albumin (P-HSA) and its receptors in hepatitis B virus (HBV) infection (1-3). Indeed, a speciesspecific receptor for P-HSA is detectable on Dane particles and on 22-nm particles of hepatitis B.Imai (2) proposed the hypothesis that P-HSA may act as a bridge between the virus and target liver cells, thus explaining the restricted host and organ tropism of HBV infection. This requires that the human hepatocyte is also provided with P-HSA receptors as has been demonstrated for rabbit hepatocytes (4). So far, human hepatocytic albumin receptors have been shown in liver sections only (5, 6). A direct demonstration of such receptors on the outer aspect of intact liver cells, however, has not been accomplished.By a micro method which permits electron microscopy of small numbers of mechanically isolated hepatocytes and utilizes albumin-coupled latex minibeads, human albumin receptors were demonstrated on human hepatocytes by phase-contrast and scanning electron microscopy (SEM).

7See acknowledgments for details Viral load measurements may predict whether human papillomavirus (HPV) type 16 infections may become persistent and eventually lead to cervical lesions. Today, multiple PCR methods exist to estimate viral load. We tested three protocols to investigate viral load as a predictor of HPV clearance. We measured viral load in 418 HPV16-positive cervical smears from 224 women participating in the Ludwig-McGill Cohort Study by low-stringency PCR (LS-PCR) using consensus L1 primers targeting over 40 known HPV types, and quantitative real-time PCR (qRT-PCR) targeting the HPV16 E6 and L1 genes. HPV16 clearance was determined by MY09/11 and PGMY PCR testing on repeated smears collected over 5 years. Correlation between viral load measurements by qRT-PCR (E6 versus L1) was excellent (Spearman's rank correlation, r50.88), but decreased for L1 qRT-PCR versus LS-PCR (r50.61). Viral load by LS-PCR was higher for HPV16 and related types independently of other concurrent HPV infections. Median duration of infection was longer for smears with high copy number by all three PCR protocols (log rank P,0.05). Viral load is inversely related to HPV16 clearance independently of concurrent HPV infections and PCR protocol.

This randomized, blinded study evaluated the immunogenicity and safety of a booster dose of Gardasil (qHPV) or Cervarix (bHPV) when administered to 12-13 year-old girls who were vaccinated at the age of 9-10 with 2 doses of qHPV (0-6 months). 366 out of 416 eligible girls participated in this follow-up study. Antibody titers were measured just before and one month post-booster. A Luminex Total IgG assay was used for antibody assessment and results are presented in Liminex Units (LU). Three years post-primary vaccination, 99-100% of subjects had detectable antibodies to 4HPV genotypes included in the qHPV with GMTs varying from 50 to 322 LU depending on genotype. After a booster dose of qHPV, a ≥4 fold increase of antibody titers to genotypes included in the vaccine was observed in 88-98% of subjects. Post-booster GMTs varied from 1666 to 4536 LU depending on genotype. These GMTs were 1.1 to 1.8-fold higher when compared to those observed one month post-second dose. After a booster of bHPV, a ≥4 fold increase of antibody titers to HPV16 and HPV18 was observed in 93-99% of subjects. The anti-HPV16 and HPV18 GMTs were 5458 and 2665 LU, respectively. These GMTs were 1.2 and 1.8 higher than those observed in the qHPV group (both P < 0.01). In bHPV group a 1.4-1.6-fold increase of antibody GMTs to HPV6 and HPV11was also observed (P < 0.001). The safety profile was acceptable for both vaccines. Both qHPV and bHPV increase antibody titers when given as a booster to girls previously vaccinated with 2 doses of qHPV. The magnitude of the immune response after booster is vaccine-dependent and has the same pattern as that reported after primary vaccination with qHPV or bHPV. When given as a booster, both vaccines have an acceptable safety profile. Longer follow-up studies are warranted to assess the need of booster doses.

SUMMARY The presence of hepatitis B surface (HBsAg) and core (HBcAg) antigens was investigated by immunofluorescence in specimens of liver tissue obtained at necrospy in 107 patients with primary hepatic carcinoma. HBsAg was detected in the cytoplasm of liver cells in 16 cases, and in eight of them the antigen was also found in malignant cells.HBcAg, which was present in the nuclei of liver cells in eight cases, was detected in the nuclei of tumour cells in six of these and also in two other cases showing HBsAg, but not HBcAg, in the nonneoplastic tissue. Although most of the primary hepatic carcinomas studied were associated with cirrhotic changes in the non-neoplastic tissue, HBsAg and HBcAg were also detected in the absence of underlying cirrhosis.Hepatitis B virus markers were demonstrated in non-neoplastic tissue, mainly in patients with a well-differentiated carcinoma, and only in these cases were they found also in the neoplastic tissue.These results show that hepatitis B virus antigens, including HBcAg, can be detected in the neoplastic cells of well-differentiated carcinoma of the liver. Although these cells could have been infected after the malignant transformation, a direct oncogenic role of the virus cannot be excluded.

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