Expression and characterization of HPV-16 L1 capsid protein in Pichia pastoris

Human papillomaviruses (HPVs) are responsible for the most common human sexually transmitted viral infections. Infection with high-risk HPVs, particularly HPV16, is associated with the development of cervical cancer. The papillomavirus L1 major capsid protein, the basis of the currently marketed vaccines, self-assembles into virus-like particles (VLPs). Here, we describe the expression, purification and characterization of recombinant HPV16 L1 produced by a methylotrophic yeast. A codon-optimized HPV16 L1 gene was cloned into a non-integrative expression vector under the regulation of a methanol-inducible promoter and used to transform competent Pichia pastoris cells. Purification of L1 protein from yeast extracts was performed using heparin-sepharose chromatography, followed by a disassembly/reassembly step. VLPs could be assembled from the purified L1 protein, as demonstrated by electron microscopy. The display of conformational epitopes on the VLPs surface was confirmed by hemagglutination and hemagglutination inhibition assays and by immuno-electron microscopy. This study has implications for the development of an alternative platform for the production of a papillomavirus vaccine that could be provided by public health programs, especially in resource-poor areas, where there is a great demand for low-cost vaccines.

Risk factors for dental erosion in a group of 12- and 16-year-old Brazilian schoolchildren

Background. Dental erosion is a multifactorial disease and is associated with dietary habits in infancy and adolescence. Aim. To investigate possible associations among dental erosion and diet, medical history and lifestyle habits in Brazilian schoolchildren. Design. The sample consisted of a random single centre cluster of 414 adolescents (12- and 16-years old) of both genders from private and public schools in Bauru (Brazil). The O`Brien [Children`s Dental Health in the United Kingdom, 1993 (1994) HMSO, London] index was used for dental erosion assessment. Data on medical history, rate and frequency of food and drinks consumption, and lifestyle habits were collected by a self-reported questionnaire. Odds ratios with 95% confidence intervals were used to assess the univariate relationships between variables. Analysis of questionnaire items was performed by multiple logistic regression analysis. The statistical significance level was set at 5%. Results. The erosion present group comprised 83 subjects and the erosion absent group 331. There were no statistically significant correlations among dental erosion and the consumption of food and drinks, medical history, or lifestyle habits. Conclusion. The results indicate that there was no correlation between dental erosion and the risk factors analysed among adolescents in Bauru/Brazil and further investigations are necessary to clarify the multifactorial etiology of this condition.

Dental Erosion in a Group of 12- and 16-year-old Brazilian Schoolchildren

Purpose: This study`s purpose was to assess the prevalence of dental erosion in adolescents and to investigate the association between erosion and sociodemographic characteristics. Methods: This study consisted of a single center cluster random sample of 414 adolescents (12 and 16 years old) of both genders from private and public schools in Bauru, Sao Paulo, Brazil. Two previously calibrated examiners (kappa=0.85) used the O`Brien (7994) index for assessment of dental erosion on the buccal and palatal surfaces of the permanent maxillary incisors and on the occlusal surfaces of the permanent first molars. Data on gender, type of school, and mean family income were collected by a questionnaire completed by the adolescents. Descriptive statistics were applied to the data, and the associations between erosion and sociodemographic variables were investigated by chi-square test (P<0.05). Results: The prevalence of dental erosion was 20% (N=83), with only enamel being involved (score 1). The labial surface was the most affected (16%). There were no significant differences between the presence of erosion and gender, type of school, and mean family income. Conclusions: The results indicate that dental erosion restricted to enamel was observed among adolescents in Bauru/Brazil, and there was no correlation between this condition and sociodemographic factors. (Pediatr Dent 201733:23-8) Received August 7, 2009 1 Last Revision October 13, 2009 I Accepted October 14, 2009