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ABSTRACT Objective The objective of this study was to clarify differences regarding HPV16 infection and gene amplification between the oral cavity and oropharynx in healthy individuals. Material and Methods The subjects were 94 healthy asymptomatic individuals (41 males, 53 females; mean age 58.6 years, range 16-97 years) who visited the Department of Oral and Maxillofacial Reconstructive Surgery of the Hiroshima University Hospital from 2014 to 2015. Oral epithelial cells were collected from oral rinse and pharynx gargle samples and placed in saline. The human endogenous retrovirus gene ERV3-1 was used as a reference to estimate the number of human cells in each sample. DNA samples were extracted from approximately 10,000 human cells and tested for HPV16 DNA by PCR using a type-specific primer. Similarly, we analyzed the HPV16 viral copy number in HPV16-positive cases using real-time PCR to examine genomic amplification. Results The percentage of HPV16-positive cases was higher in the gargle (28.7%) as compared to the rinse (16.0%) samples. In the oral rinse samples, males (26.8%) showed a significantly higher rate of HPV16 than females (7.5%) (P=0.021). Importantly, in older subjects (aged 60-89 years), gargle samples showed a significantly higher rate of HPV16 (33.3%) than oral rinse samples (13.7%) (P=0.034). The average number of viral copies was approximately 8 times higher in the gargle than in the oral rinse samples (0.16±0.27 vs. 1.35±1.26 copy numbers per cell), a significant difference (P<0.001). Conclusion Our findings suggest that the oropharynx is more susceptible to HPV16 infection as compared to the oral cavity, while HPV16 gene amplification is also more commonly found in the oropharynx.
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Humanos , Masculino , Femenino , Adolescente , Adulto , Persona de Mediana Edad , Anciano , Anciano de 80 o más Años , Adulto Joven , Orofaringe/virología , Amplificación de Genes/fisiología , Infecciones por Papillomavirus/epidemiología , Infecciones por Papillomavirus/virología , Papillomavirus Humano 16/genética , Boca/virología , Factores de Tiempo , ADN Viral , Recuento de Células , Prevalencia , Factores de Riesgo , Factores de Edad , Variaciones en el Número de Copia de ADN , Reacción en Cadena en Tiempo Real de la Polimerasa , Japón/epidemiologíaRESUMEN
La incidencia de candidiasis sistémica se ha incrementado en asociación a la sobrevida de pacientes neutropénicos. El diagnóstico precoz presenta limitaciones debido a la baja sensibilidad y especificidad de los métodos diagnósticos en uso; esto explica que el diagnóstico y tratamiento se basan en muchos casos en la sospecha clínica. El tratamiento de la candidiasis sistémica no es siempre exitoso y presenta una alta incidencia de reacciones adversas y complicaciones. En la actualidad, la profilaxis constituye la medida más importante para disminuir la morbimortalidad de eta infección, sin embargo, para establecer medidas eficaces de control, es necesario ahondar en la epidemiología de las infecciones por Candida spp, lo que requiere de sistemas adecuados de caracterización de cepas involucradas. Este estudio presenta los resultados obtenidos mediante el procedimiento denominado amplificación aleatoria de fragmentos de DNA (RAPD-PCR) utilizado como un método de análisis espidemiológico molecular
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Candida albicans/genética , Técnicas In Vitro , Amplificación de Genes/fisiología , Análisis de Secuencia de ADN/métodos , Candida albicans/patogenicidad , Candidiasis/microbiologíaRESUMEN
The discovery of Dna sequence amplification in sciarid flies and investigations into its control and biological significance are reviewed. Results thus far show that amplification of specific salivary gland polytene chromosome bands is a general phenomenon in sciarids. It brought about as part of a final endoreplication cycle by the rising titer of ecdysterone that occurs as the Larvae approach the prepupal period. Amplification and transcription of these bands is a late, multistep effect of this hormone.The Dna puffs which form in amplified region produce mRNAs which are translated into polypeptides that appear to be involved in coccon formation. Application of molecular cloning techniques to the study of Dna amplification has allowed precise quantitation of amplification for several Dna puffs and is yielding maps of their transcription units.These techniques will ultimately help to define the origins of Dna puff replication and contribute to an understanding of the mechanism and control of the amplification phenomenon in sciaridae. Projections for future experimental approaches are presented