Development and validation of a French questionnaire that assesses knowledge, attitude, and practices toward Marburg diseases in sub-Saharan African countries.

OBJECTIVES: Marburg virus, previously referred to as Marburg hemorrhagic fever, is a highly severe and frequently fatal illness that affects humans. This study aimed to develop and validate a French questionnaire to assess knowledge, attitude, and practice toward Marburg virus disease (FKAP-MVD). STUDY DESIGN: An anonymous online survey was used, which was distributed through various platforms and emails. Data were collected from Burkina Faso, Guinea, the Democratic Republic of Congo, and Senegal. METHODS: To conduct the study, an anonymous online survey was used, which was distributed through various platforms such as Facebook, Twitter, WhatsApp, and emails. The survey was uploaded onto a Google form to facilitate data collection. Data were collected from Burkina Faso, Guinea, the Democratic Republic of Congo, and Senegal. RESULTS: Of the total sample of 510 participants, 60.0% were male, their mean age was 28.41 ± 6.32 years, 38.0% were married, 86.6% resided in urban areas and 64.1% had a university education. The questionnaire had good internal consistency; Cronbach's alpha was 0.87. The correlation between knowledge and attitude was 0.002, the correlation between knowledge and practice was 0.204, and the correlation between practice and attitude was relatively weak and negative at -0.060. This indicates the divergent validity of the questionnaire. The KMO value of 0.91 indicates a high level of adequacy, suggesting that the data are suitable for factor analysis. The Bartlett test of Sphericity yielded an approximate χ2 value of 4016.890 with 300 degrees of freedom and a P-value of 0.0001. The confirmatory factor analysis revealed 25 questions in three domains. The normed chi-square value is 1.224. The goodness of Fit Index (GFI) is 0.902, the Comparative Fit Index (CFI) is 0.982, the Root Mean Square Error of Approximation (RMSEA) is 0.033, and the Root Mean Square Residual (RMR) is 0.062. These values indicate a good fit of the model to the data. CONCLUSIONS: In general, the developed questionnaire has significant potential to inform public health initiatives and interventions related to MVD.

Porphyromonas gingivalis Impairs Oral Epithelial Barrier through Targeting GRHL2.

Oral mucosa provides the first line of defense against a diverse array of environmental and microbial irritants by forming the barrier of epithelial cells interconnected by multiprotein tight junctions (TJ), adherens junctions, desmosomes, and gap junction complexes. Grainyhead-like 2 (GRHL2), an epithelial-specific transcription factor, may play a role in the formation of the mucosal epithelial barrier, as it regulates the expression of the junction proteins. The current study investigated the role of GRHL2 in the Porphyromonas gingivalis (Pg)-induced impairment of epithelial barrier functions. Exposure of human oral keratinocytes (HOK-16B and OKF6 cells) to Pg or Pg-derived lipopolysaccharides (Pg LPSs) led to rapid loss of endogenous GRHL2 and the junction proteins (e.g., zonula occludens, E-cadherin, claudins, and occludin). GRHL2 directly regulated the expression levels of the junction proteins and the epithelial permeability for small molecules (e.g., dextrans and Pg bacteria). To explore the functional role of GRHL2 in oral mucosal barrier, we used a Grhl2 conditional knockout (KO) mouse model, which allows for epithelial tissue-specific Grhl2 KO in an inducible manner. Grhl2 KO impaired the expression of the junction proteins at the junctional epithelium and increased the alveolar bone loss in the ligature-induced periodontitis model. Fluorescence in situ hybridization revealed increased epithelial penetration of oral bacteria in Grhl2 KO mice compared with the wild-type mice. Also, blood loadings of oral bacteria (e.g., Bacteroides, Bacillus, Firmicutes, ß-proteobacteria, and Spirochetes) were significantly elevated in Grhl2 KO mice compared to the wild-type littermates. These data indicate that Pg bacteria may enhance paracellular penetration through oral mucosa in part by targeting the expression of GRHL2 in the oral epithelial cells, which then impairs the epithelial barrier by inhibition of junction protein expression, resulting in increased alveolar tissue destruction and systemic bacteremia.

Human Papillomavirus 16 E6 Induces FoxM1B in Oral Keratinocytes through GRHL2.

High-risk human papillomavirus (HPV) is a major risk factor for oral and pharyngeal cancers (OPCs), yet the detailed mechanisms by which HPV promotes OPCs are not understood. Forkhead box M1B (FoxM1B) is an oncogene essential for cell cycle progression and tumorigenesis, and it is aberrantly overexpressed in many tumors. We previously showed that FoxM1B was the putative target of an epithelial-specific transcription factor, Grainyhead-like 2 (GRHL2). In the current study, we demonstrate that HPV type 16 (HPV-16) E6 induces FoxM1B in human oral keratinocytes (HOKs) and tonsillar epithelial cells (TECs) in part through GRHL2. FoxM1B was barely detectable in cultured normal human oral keratinocytes (NHOKs) and progressively increased in immortalized HOKs harboring HPV-16 genome (HOK-16B) and tumorigenic HOK-16B/BaP-T cells. Retroviral expression of HPV-16 E6 and/or E7 in NHOKs, TECs, and hypopharyngeal carcinoma cells (FaDu) revealed induction of FoxM1B and GRHL2 by the E6 protein but not E7. Both GRHL2 and FoxM1B were strongly induced in the epidermis of HPV-16 E6 transgenic mice and HPV+ oral squamous cell carcinomas. Ectopic expression of FoxM1B led to acquisition of transformed phenotype in HOK-16B cells. Loss of FoxM1B by lentiviral short hairpin RNA vector or chemical inhibitor led to elimination of tumorigenic characteristics of HOK-16B/BaP-T cells. Luciferase reporter assay revealed that GRHL2 directly bound and regulated the FoxM1B gene promoter activity. Using epithelial-specific Grhl2 conditional knockout mice, we exposed wild-type (WT) and Grhl2 KO mice to 4-nitroquinolin 1-oxide (4-NQO), which led to induction of FoxM1B in the tongue tissues and rampant oral tumor development in the WT mice. However, 4-NQO exposure failed to induce tongue tumors or induction of FoxM1B expression in Grhl2 KO mice. Collectively, these results indicate that HPV-16 induces FoxM1B in part through GRHL2 transcriptional activity and that elevated FoxM1B level is required for oropharyngeal cancer development.