BackgroundMicrobial dysbiosis and microbiome-induced inflammation have emerged as important factors in oral squamous cell carcinoma (OSCC) tumorigenesis during the last two decades. However, the “rare biosphere” of the oral microbiome, including fungi, has been sparsely investigated. This study aimed to characterize the salivary mycobiome in a prospective Sudanese cohort of OSCC patients and to explore patterns of diversities associated with overall survival (OS).Materials and MethodsUnstimulated saliva samples (n = 72) were collected from patients diagnosed with OSCC (n = 59) and from non-OSCC control volunteers (n = 13). DNA was extracted using a combined enzymatic–mechanical extraction protocol. The salivary mycobiome was assessed using a next-generation sequencing (NGS)-based methodology by amplifying the ITS2 region. The impact of the abundance of different fungal genera on the survival of OSCC patients was analyzed using Kaplan–Meier and Cox regression survival analyses (SPPS).ResultsSixteen genera were identified exclusively in the saliva of OSCC patients. Candida, Malassezia, Saccharomyces, Aspergillus, and Cyberlindnera were the most relatively abundant fungal genera in both groups and showed higher abundance in OSCC patients. Kaplan–Meier survival analysis showed higher salivary carriage of the Candida genus significantly associated with poor OS of OSCC patients (Breslow test: p = 0.043). In contrast, the higher salivary carriage of Malassezia showed a significant association with favorable OS in OSCC patients (Breslow test: p = 0.039). The Cox proportional hazards multiple regression model was applied to adjust the salivary carriage of both Candida and Malassezia according to age (p = 0.029) and identified the genus Malassezia as an independent predictor of OS (hazard ratio = 0.383, 95% CI = 0.16–0.93, p = 0.03).ConclusionThe fungal compositional patterns in saliva from OSCC patients were different from those of individuals without OSCC. The fungal genus Malassezia was identified as a putative prognostic biomarker and therapeutic target for OSCC.
Efficient seed dispersal in flowering plants is enabled by the development of fruits, which can be either dehiscent or indehiscent. Dehiscent fruits open at maturity to shatter the seeds, while indehiscent fruits do not open and the seeds are dispersed in various ways. The diversity in fruit morphology and seed shattering mechanisms is enormous within the flowering plants. How these different fruit types develop and which molecular networks are driving fruit diversification is still largely unknown, despite progress in eudicot model species. The orchid family, known for its astonishing floral diversity, displays a huge variation in fruit dehiscence types, which have been poorly investigated. We undertook a combined approach to understand fruit morphology and dehiscence in different orchid species to get more insight into the molecular network that underlies orchid fruit development. We describe fruit development in detail for the epiphytic orchid species Erycina pusilla and compare it to two terrestrial orchid species: Cynorkis fastigiata and Epipactis helleborine . Our anatomical analysis provides further evidence for the split carpel model, which explains the presence of three fertile and three sterile valves in most orchid species. Interesting differences were observed in the lignification patterns of the dehiscence zones. While C. fastigiata and E. helleborine develop a lignified layer at the valve boundaries, E. pusilla fruits did not lignify at these boundaries, but formed a cuticle-like layer instead. We characterized orthologs of fruit-associated MADS-domain transcription factors and of the Arabidopsis dehiscence-related genes INDEHISCENT (IND)/HECATE 3 (HEC3), REPLUMLESS (RPL) and SPATULA (SPT)/ALCATRAZ (ALC) in E. pusilla , and found that the key players of the eudicot fruit regulatory network appear well-conserved in monocots. Protein-protein interaction studies revealed that MADS-domain complexes comprised of FRUITFULL (FUL), SEPALLATA (SEP) and AGAMOUS (AG) /SHATTERPROOF (SHP) orthologs can also be formed in E. pusilla , and that the expression of HEC3, RPL , and SPT can be associated with dehiscence zone development similar to Arabidopsis. Our expression analysis also indicates differences, however, which may underlie fruit divergence.
Background: Tumor immune infiltrate has been explored in oral squamous cell carcinoma (OSCC), but studies on simultaneous characterization of multiple immune cell subtypes separately in stromal and intraepithelial tumor compartments are limited.Objectives: We aimed to investigate the immune cell infiltrate in OSCC by using immunohistochemistry (IHC) for a panel of inflammatory cells in stromal and epithelial tumor compartments for a better characterization of the tumors. Methods: Thirty-six OSCC lesions and nine normal oral mucosa (NOM) samples from patients attending Khartoum Dental Teaching Hospital, Sudan were investigated for presence of tumor infiltrating lymphocytes, tumor-associated macrophages, tumorassociated neutrophils, and PD-L1 positive cells in the inflammatory infiltrate by single and double IHC. Digital quantitative analysis (Aperio Technologies Inc.) was performed separately for stromal and epithelial compartments.Results: OSCC cases displayed a higher inflammatory infiltrate in the associated stroma, but not in the epithelial compartment when compared to NOM. The immunosuppressive type of inflammatory infiltrate, that is, T regulatory cells (FoxP3 + cells) was identified to be significantly higher in the epithelial compartment of tumors with Elisabeth Sivy Nginamau and Daniela Elena Costea contributed equally to this work.
Background Although several markers have been used for enrichment of cells with stem cell‐like properties in oral squamous cell carcinoma (OSCC), isolation of a pure subpopulation is still a challenging task. Normal oral and esophageal keratinocyte stem cells have been previously isolated using the low‐affinity nerve growth factor receptor p75NTR. Objective To investigate the potential of p75NTR as a marker for identification and isolation of oral cancer cells with stem cell‐like properties. Methods Subpopulations of cells with high or low expression of p75NTR were sorted from OSCC‐derived cells and compared for sphere/colony formation, in vivo tumor formation ability, expression of stem cell‐related molecules, cell cycle distribution and drug resistance. Results p75NTRHigh cells exhibited statistically significant higher stem cell properties than p75NTRLow cells in all assays performed. Nevertheless, p75NTRLow subpopulation did also exhibit some stem cell features, but to a lesser extent. Propagation of p75NTRLow cells for several passages in culture showed that the expression of p75NTR could rise spontaneously. This finding was also supported by the similar expression of p75NTR by the xenografts generated by both subpopulations in NOD\SCID IL2Rgnullmice. Conclusion p75NTR can be used for isolating a subpopulation enriched for cells with stem cell‐like properties in OSCC. De novo generation of p75NTRHigh cells from p75NTRLow cells suggests either that there is another subpopulation with stem cell features within the p75NTRLow cells, or that the p75NTRLow cells can dedifferentiate due to a contextually regulated equilibrium between stem cell‐like cells and transit‐amplifying neoplastic progenitors.
Objective: to determine the agreement between the conventional and digital radiographs in detection of intra canal separated files. Material & Methods: A cross sectional study was conducted at Operative Dentistry Department, Institute of Dentistry, Liaquat University of Medical and Health Sciences, Jamshoro from 01-03-2020 to 31-08-2020. A total of 71 Permanent single straight rooted, consisting of single canal extracted teeth were included in this study. Preoperatively radiograph was taken from mesiodistal (M-D) and buccolingual (B-L) direction to confirm the inclusion criteria. Root canal was prepared and the file was inserted to the working length, a counterclockwise motion was performed till the separation of file occurs. Then radiograph was taken to confirm the separated file at working length. Results: Agreement between the conventional and digital radiographs in detection of intra canal separated files was 90.4%. The kappa value between the conventional and digital radiographs was 0.534 which is statistically significant and considered as a moderate degree of agreement. Practical Implication: Correct radiographic detection helps identify the cause, improves in therapy choice, and offers medico legal security in cases of retreatment. Conclusion: In conclusion, both modalities can be used for detection and facilitate in finding the cause and helps in decision making for treatment and provides medico legal security in the retreatment cases. Keywords: Conventional radiograph, Digital Radiographs, Endodontic treatment, Separated file.
Expression of cancer-associated biomarkers represents a great potential for a more precise diagnosis for patient stratification and more personalized treatment of oral squamous cell carcinoma (OSCC). The aim of this study was to develop an immunohistochemistry-based malignancy index diagnostic system (IHC-MIDS) based on the expression of a panel of several molecules expressed by different tumor components and involved in different oncogenic pathways. IHC was performed on paraffin embedded formalin fixed tissue samples from 102 patients for the following biomarkers: p16, p53, ki67, p75NTR, LoxL4, aSMA, Foxp3, CD68, CD 163, CD80, FVIII and D2-40. The stained tissues were scanned and quantified using Immage J open source software. An IHC-MIDS score was developed using correlation-based feature selection algorithm. IHC-MIDS was able to stratify OSCC lesions for several clinical pathological features, including the risk of recurrence. This study shows that a scoring system based on expression of a panel of biomarkers is more precise than the use of a single biomarker and that it can be used for risk stratification of OSCC lesions in order to guide further management. Citation Format: Martha Rolland Jacobsen, Harsh Dongre, Israa Ahmed, Vidisha Tuljaurkar, Prathamesh S. Pai, Asawari Patil, Dipak Sapkota, Anne Christine Johannessen, Nenad Filipovic, Milind Vaidya, Sharda Sawant, Daniela Elena Costea. Development of a molecular diagnostic tool for more precise diagnosis of oral squamous cell carcinoma [abstract]. In: Proceedings of the AACR-AHNS Head and Neck Cancer Conference: Optimizing Survival and Quality of Life through Basic, Clinical, and Translational Research; April 23-25, 2017; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2017;23(23_Suppl):Abstract nr 45.
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