Diversity of Anal HPV and Non-HPV Sexually Transmitted Infections and Concordance with Genital Infections in HIV-Infected and HIV-Uninfected Women in the Tapajós Region, Amazon, Brazil.

The aim of this study was to classify the diversity of anal HPV and non-HPV sexually transmitted infections (STIs) and compare the concordance between anal and genital infections in HIV-infected and uninfected women living in the Tapajós region, Amazon, Brazil. A cross-sectional study was performed with 112 HIV-uninfected and 41 HIV-infected nonindigenous women. Anal and cervical scrapings were collected and analyzed for HPV, Chlamydia trachomatis (CT), Neisseria gonorrheae (NG), Trichomonas vaginalis (TV), Mycoplasma genitalium (MG), and Human alphaherpesvirus 2 (HSV-2). The Kappa test evaluated the concordance between anal and genital infections. The overall prevalence of anal HPV infection was 31.3% in HIV-uninfected and 97.6% in HIV-infected women. The most frequent anal high-risk HPV (hrHPV) types were HPV18 and HPV16 in HIV-uninfected women and HPV51, HPV59, HPV31, and HPV58 in HIV-infected women. Anal HPV75 Betapapillomavirus was also identified. Anal non-HPV STIs were identified in 13.0% of all participants. The concordance analysis was fair for CT, MG, and HSV-2, almost perfect agreement for NG, moderate for HPV, and variable for the most frequent anal hrHPV types. Thus, a high prevalence of anal HPV infection with moderate and fair concordance between anal and genital HPV and non-HPV STIs was observed in our study.

Relação entre desmame precoce, desenvolvimento de hábitos bucais deletérios e maloclusões na infância / Relationship between early weaning, development of harmful oral habits and malocclusions in childhood

Objetivo: o objetivo desta pesquisa foi analisar a associação do tempo de aleitamento materno com a prática de hábitos deletérios e o desenvolvimento de maloclusões na primeira infância. Materiais e Métodos: foram aplicados 171 formulários às mães com 18 anos ou mais que acompanharam o processo de amamentação de seu(s) filho(s) de 4 a 10 anos de idade. O questionário foi composto por 22 perguntas, sendo 15 específicas e relacionadas ao aleitamento, hábitos deletérios da criança (uso de chupeta, sucção de dedo e sucção do próprio lábio) e maloclusões específicas (mordida cruzada posterior, mordida cruzada anterior e mordida aberta). Resultados: a prevalência da amamentação exclusiva por 6 meses ou mais foi de 52%, enquanto 29,8% das crianças foram amamentadas de 0 a 5 meses e 18,2% nunca foram amamentadas no seio exclusivamente. Quanto a alimentação complementar, grande parcela (37,4%) encontrada evidenciou início após os 6 meses; 57,9% das mães relataram algum tipo de dificuldade para amamentar e 34,5% das crianças fizeram uso de chupeta. Em relação aos hábitos deletérios, 10% desenvolveram sucção de dedo. O relato de desenvolvimento de maloclusões foi de 9,4% das crianças com mordida cruzada posterior; 7,6% mordida cruzada anterior e 18,7% mordida aberta anterior. Conclusão: sendo assim, é possível inferir que o tempo preconizado pela Organização Mundial da Saúde para aleitamento materno exclusivo foi fundamental para o não desenvolvimento de hábitos deletérios e maloclusões. Em contrapartida, quanto mais cedo a introdução de alimentos complementares, e a interrupção do aleitamento exclusivo nos seis primeiros meses, maior o risco do desenvolvimento de hábitos e consequentemente maloclusões.

Objective: the objective of this research was to analyze the association of breastfeeding duration with the practice of deleterious habits and the development of malocclusions in early childhood. Materials and Methods: a total of 171 forms were applied to mothers aged 18 years or older who followed the breastfeeding process of their child(ren) between 4 and 10 years of age. The questionnaire consisted of 22 questions, 15 of which were specific and related to breastfeeding, the child's deleterious habits (use of a pacifier, finger sucking and lip sucking) and specific malocclusions (posterior crossbite, anterior crossbite and open bite). Results: the prevalence of exclusive breastfeeding for 6 months or more was 52%, while 29.8% of children were breastfed from 0 to 5 months and 18.2% were never exclusively breastfed. As for complementary feeding, a large portion (37.4%) found to start after 6 months; 57.9% of the mothers reported some type of difficulty in breastfeeding and 34.5% of the children used a pacifier. Regarding deleterious habits, 10% developed finger sucking. The report of development of malocclusions was 9.4% of children with posterior crossbite; 7.6% anterior crossbite and 18.7% anterior open bite. Conclusion: therefore, it is possible to infer that the time recommended by the World Health Organization for exclusive breastfeeding was fundamental for the non-development of deleterious habits and malocclusions. On the other hand, the earlier the introduction of complementary foods and the interruption of exclusive breastfeeding in the first six months, the greater the risk of developing habits and, consequently, malocclusions.

A novel evaluation method for Ki-67 immunostaining in paraffin-embedded tissues.

The Ki-67 labeling index is traditionally used to investigate tumor aggressiveness. However, no diagnostic or prognostic value has been associated to the heterogeneous pattern of nuclear positivity. The aims of this study were to develop a classification for the patterns of Ki-67-positive nuclei; to search scientific evidence for the Ki-67 expression and location throughout the cell cycle; and to develop a protocol to apply the classification of patterns of Ki-67-positive nuclei in squamous epithelium with different proliferative activities. Based on empirical observation of paraffin sections submitted to immunohistochemistry for the determination of Ki-67 labeling index and literature review about Ki-67 expression, we created a classification of the patterns of nuclear positivity (NP1, NP2, NP3, NP4, and mitosis). A semi-automatic protocol was developed to identify and quantify the Ki-67 immunostaining patterns in target tissues. Two observers evaluated 7000 nuclei twice to test the intraobserver reliability, and six evaluated 1000 nuclei to the interobserver evaluation. The results showed that the immunohistochemical patterns of Ki-67 are similar in the tumoral and non-tumoral epithelium and were classified without difficulty. There was a high intraobserver reliability (Spearman correlation coefficient > 0.9) and moderate interobserver agreement (k = 0.523). Statistical analysis showed that non-malignant epithelial specimens presented a higher number of NP1 (geographic tongue = 83.8 ± 21.8; no lesion = 107.6 ± 52.7; and mild dysplasia = 86.6 ± 25.8) when compared to carcinoma in Situ (46.8 ± 34.8) and invasive carcinoma (72.6 ± 37.9). The statistical evaluation showed significant difference (p < 0.05). Thus, we propose a new way to evaluate Ki-67, where the pattern of its expression may be associated with the dynamics of the cell cycle. Future proof of this association will validate the use of the classification for its possible impact on cancer prognosis and guidance on personalized therapy.

Hybridization Capture-Based Next-Generation Sequencing to Evaluate Coding Sequence and Deep Intronic Mutations in the NF1 Gene.

Neurofibromatosis 1 (NF1) is one of the most common genetic disorders and is caused by mutations in the NF1 gene. NF1 gene mutational analysis presents a considerable challenge because of its large size, existence of highly homologous pseudogenes located throughout the human genome, absence of mutational hotspots, and diversity of mutations types, including deep intronic splicing mutations. We aimed to evaluate the use of hybridization capture-based next-generation sequencing to screen coding and noncoding NF1 regions. Hybridization capture-based next-generation sequencing, with genomic DNA as starting material, was used to sequence the whole NF1 gene (exons and introns) from 11 unrelated individuals and 1 relative, who all had NF1. All of them met the NF1 clinical diagnostic criteria. We showed a mutation detection rate of 91% (10 out of 11). We identified eight recurrent and two novel mutations, which were all confirmed by Sanger methodology. In the Sanger sequencing confirmation, we also included another three relatives with NF1. Splicing alterations accounted for 50% of the mutations. One of them was caused by a deep intronic mutation (c.1260 + 1604A > G). Frameshift truncation and missense mutations corresponded to 30% and 20% of the pathogenic variants, respectively. In conclusion, we show the use of a simple and fast approach to screen, at once, the entire NF1 gene (exons and introns) for different types of pathogenic variations, including the deep intronic splicing mutations.

Estudo da resposta imune em lesões intraepiteliais anais em pacientes infectados e não infectados pelo HIV e HPV / Study of immune response in intraepithelial lesions in infected and non-infected patients by HIV and HPV

O Papiloma Vírus Humano (HPV) é o principal agente etiológico do câncer do trato anogenital. Uma maior prevalência e incidência de desenvolvimento do carcinoma e doenças associadas ao HPV têm sido observadas em indivíduos infectados pelo HIV. A história natural da infecção pelo HPV ainda não está totalmente elucidada, assim como aresposta imune que ocorre na co-infeção pelo HIV/HPV,principalmente na mucosa anal.Objetivo do presente estudo foi avaliar a resposta imune “in situ” em amostras de biópsias de indivíduos infectados pelo HIV em acompanhamento no Instituto Nacional de Infectologia/FIOCRUZ, RJ. Um total de 114 biópsias foi analisado através de TissueMicro-Array, sendo 15 de indivíduos Não infectados pelo HIV, todos sem lesão, e 99 deindivíduos Infectados pelo HIV: 21 sem lesão, 39 com NIA1, e 39 com NIA2/3. Foirealizado PCR e sequenciamento para identificação do DNA de HPV e imunohistoquímicapara análise dos marcadores imunes CD4, CD8, Foxp3, T-bet e IL-10 e SLPI. A análiseestatística foi feita no software SPSS 15.0 aplicando os testes: Kruskall-Wallis, Teste Quiquadradoe Teste Exato de Fisher. Pacientes Infectados pelo HIV com NIA 2/3apresentaram nadir CD4+ <50 cél/mm³ comparados aos pacientes normais (p = 0,01).Quanto aos marcadores imunes, indivíduos Infectados pelo HIV apresentaram uma maiorexpressão de Foxp3 e IL-10 de acordo com a gravidade da lesão (p= 0,002)...

Human Papillomavirus (HPV) is the main etiologic agent for lower genital tract cancers. Ahigher incidence and prevalence of these cancers has been reported in HIV infectedindividuals. The natural history of HPV infection is still unclear and the immune responsewith HIV/HPV co-infection, particularly in the anal mucosa, is poorly understood. The aimof this study was to evaluate the immune response in situ in ano-rectal biopsies fromHIV-infected patients followed up at the National Institute of Infectious Diseases /FIOCRUZ, RJ. A total of 114 biopsies were analyzed by Tissue Micro-Array: 15 were fromHIV negative individuals without any lesions and 99 from HIV-positive individuals: 21without lesions, 39 with AIN 1 and 39 with AIN 2/3. PCR and sequencing was run toidentify HPV DNA. CD4, CD8, Foxp3, T-bet, IL-10 and SLPI were analyzed byimmunohistochemistry. Statistical analysis was carried out using SPSS 15.0. TheKruskal-Wallis, chi-square and Fisher's exact tests were applied. HIV-positive patients withAIN 2/3 showed a nadir count of CD4 + <50 cells/mm³ compared to normal subjects (p =0.01). HIV positive individuals showed a higher expression of FoxP3 and IL-10 accordingto the severity of the lesion (p = 0.002). A positive coefficient correlation was foundbetween FoxP3 and IL-10 (0.34; p = 0.27). The analysis showed that 93.4 percent (101/107) ofthe samples had HPV DNA, and the most common types were: HPV 16 (26.9 percent), followedby HPV 6 (15.7 percent), HPV 59 (13 percent) and HPV 18 (10.2 percent). Interestingly, samples fromindividuals with high-risk oncogenic HPV DNA were negative for SLPI and there was lessexpression in AIN 2/3 compared to the no-lesion group of HIV + individuals, showing aninverse association with HPV type and degree of AIN...

Dysphonia as a sign of HPV laryngeal infection: a case report.

BACKGROUND: Voice dysfunction or dysphonia may be associated with several clinical conditions. Among these, laryngeal human papillomavirus (HPV)-induced lesions should be considered as a possible causative factor. We report a case of dysphonia in a patient presenting with an HPV laryngeal lesion. We also discuss the clinical features of the disease, its histopathological findings, and treatment and rigorous follow-up. CASE PRESENTATION: We report a case of laryngeal papilloma in a 29-year-old, Afro-descendant, male patient with dysphonia. He was a non-smoker and was not a drug user. Videolaryngostroboscopy revealed signs suggestive of pharyngolaryngeal reflux. The right vocal fold presented with a papillomatous aspect in the posterior third, which underwent excision. Histopathological examination showed a nodular lesion of the right vocal fold, conclusive of squamous papilloma with absence of malignancy. CONCLUSION: Patients presenting with persistent voice dysfunction or dysphonia should be investigated for possible laryngeal HPV infection. Diagnostic confirmation by HPV genotyping is important for follow-up of potential recurrence.