ABSTRACT
Current estimates suggest 50% of glaucoma blindness worldwide is caused by primary angle-closure glaucoma (PACG) but the causative gene is not known. We used genetic linkage and whole genome sequencing to identify Spermatogenesis Associated Protein 13, SPATA13 (NM_001166271; NP_001159743, SPATA13 isoform I), also known as ASEF2 (Adenomatous polyposis coli-stimulated guanine nucleotide exchange factor 2), as the causal gene for PACG in a large seven-generation white British family showing variable expression and incomplete penetrance. The 9 bp deletion, c.1432_1440del; p.478_480del was present in all affected individuals with angle-closure disease. We show ubiquitous expression of this transcript in cell lines derived from human tissues and in iris, retina, retinal pigment and ciliary epithelia, cornea and lens. We also identified eight additional mutations in SPATA13 in a cohort of 189 unrelated PACS/PAC/PACG samples. This gene encodes a 1277 residue protein which localises to the nucleus with partial co-localisation with nuclear speckles. In cells undergoing mitosis SPATA13 isoform I becomes part of the kinetochore complex co-localising with two kinetochore markers, polo like kinase 1 (PLK-1) and centrosome-associated protein E (CENP-E). The 9 bp deletion reported in this study increases the RAC1-dependent guanine nucleotide exchange factors (GEF) activity. The increase in GEF activity was also observed in three other variants identified in this study. Taken together, our data suggest that SPATA13 is involved in the regulation of mitosis and the mutations dysregulate GEF activity affecting homeostasis in tissues where it is highly expressed, influencing PACG pathogenesis.
Subject(s)
Glaucoma, Open-Angle/genetics , Guanine Nucleotide Exchange Factors/genetics , Mutation , Adolescent , Adult , Aged , Cell Division , Cell Nucleus/metabolism , Eye/metabolism , Female , Glaucoma, Open-Angle/pathology , Guanine Nucleotide Exchange Factors/chemistry , Guanine Nucleotide Exchange Factors/metabolism , Humans , Kinetochores/metabolism , Male , Middle Aged , Pedigree , Protein Isoforms/chemistry , Protein Isoforms/genetics , Protein Isoforms/metabolism , Protein TransportABSTRACT
Vimentin, a type III intermediate filament protein, is found in most cells along with microfilaments and microtubules. It has been shown that the head domain folds back to associate with the rod domain and this association is essential for filament assembly. The N-terminally tagged vimentin has been widely used to label the cytoskeleton in live cell imaging. Although there is previous evidence that EGFP tagged vimentin fails to form filaments but is able to integrate into a pre-existing network, no study has systematically investigated or established a molecular basis for this observation. To determine whether a tag would affect de novo filament assembly, we used vimentin fused at the N-terminus with two different sized tags, AcGFP (239 residues, 27 kDa) and 3 × FLAG (22 residues; 2.4 kDa) to assemble into filaments in two vimentin-deficient epithelial cells, MCF-7 and A431. We showed that regardless of tag size, N-terminally tagged vimentin aggregated into globules with a significant proportion co-aligning with ß-catenin at cell-cell junctions. However, the tagged vimentin aggregates could form filaments upon adding untagged vimentin at a ratio of 1:1 or when introduced into cells containing pre-existing filaments. The resultant filament network containing a mixture of tagged and untagged vimentin was less stable compared to that formed by only untagged vimentin. The data suggest that placing a tag at the N-terminus may create steric hinderance in case of a large tag (AcGFP) or electrostatic repulsion in case of highly charged tag (3 × FLAG) perhaps inducing a conformational change, which deleteriously affects the association between head and rod domains. Taken together our results shows that a free N-terminus is essential for filament assembly as N-terminally tagged vimentin is not only incapable of forming filaments, but it also destabilises when integrated into a pre-existing network.
Subject(s)
Cytoskeleton , Intermediate Filaments , Amino Acid Sequence , Cytoskeleton/metabolism , Intermediate Filaments/metabolism , Transfection , Vimentin/metabolismABSTRACT
BACKGROUND: The concept of head and neck cancers (HNSCC) having unique molecular signatures is well accepted but relating this to clinical presentation and disease behaviour is essential for patient benefit. Currently the clinical significance of HNSCC molecular subtypes is uncertain therefore personalisation of HNSCC treatment is not yet possible. METHODS: We performed meta-analysis on 8 microarray studies and identified six significantly up- (PLAU, FN1, CDCA5) and down-regulated (CRNN, CLEC3B and DUOX1) genes which were subsequently quantified by RT-qPCR in 100 HNSCC patient margin and core tumour samples. RESULTS: Retrospective correlation with sociodemographic and clinicopathological patient details identified two subgroups of opposing molecular signature (+q6 and -q6) that correlated to two recognised high-risk HNSCC populations in the UK. The +q6 group were older, male, and excessive alcohol users whilst the -q6 group were younger, female, paan-chewers and predominantly Bangladeshi. Additionally, all patients with tumour recurrence were in the latter subgroup. CONCLUSIONS: We provide the first evidence linking distinct molecular signatures in HNSCC with clinical presentations. Prospective trials are required to determine the correlation between these distinct genotypes and disease progression or treatment response. This is an important step towards the ultimate goal of improving outcomes by utilising personalised molecular-signature-guided treatments for HNSCC patients.
Subject(s)
Squamous Cell Carcinoma of Head and Neck/etiology , Squamous Cell Carcinoma of Head and Neck/metabolism , Biomarkers, Tumor , Computational Biology/methods , Data Mining , Disease Susceptibility , Female , Gene Expression Profiling , Genetic Predisposition to Disease , Humans , Kaplan-Meier Estimate , Male , Neoplasm Staging , Prognosis , Retrospective Studies , Selection, Genetic , Squamous Cell Carcinoma of Head and Neck/diagnosis , Squamous Cell Carcinoma of Head and Neck/mortality , TranscriptomeABSTRACT
Introduction: Pediatric oral pathology encompasses a wide range of diseases and conditions affecting the oral cavity in children. In this study we conducted an analysis on a dataset from one academic center in Saudi Arabia to investigate the prevalence of various oral lesions in pediatric patients. Methods: To conduct this study we analyzed oral pathology reports of pediatric patients (age range: 0-18 years) who underwent biopsies at our institution's oral pathology laboratory between January 2012 and December 2022. The data we collected included demographic information and diagnostic findings. Statistical analysis was performed to determine the prevalence rates of and associations between different variables. Results: The study analyzed a total of 183 pediatric cases with oral pathologies. The most common conditions observed were mucocele (12 %), periapical granuloma (7.7 %), dentigerous cyst (6.6 %), and radicular cyst (6.6 %). Some conditions showed variations in prevalence based on age and gender. For example, mucoceles were more prevalent among patients who were 6-12 years of age and female. Conclusion: This retrospective study provides valuable insights into the prevalence and demographic characteristics of pediatric oral pathologies. By understanding the prevalence of conditions in this population and recognizing differences in distribution compared with those cited in other studies, we highlighted the importance of considering regional and demographic influences. Further prospective studies are needed to investigate factors that may contribute to these variations.
ABSTRACT
BACKGROUND: Oral-pigmented lesions have raised aesthetic concerns, leading to multiple depigmentation techniques. This systematic review and network meta-analysis aimed to assess the efficacy of different treatments for oral hyperpigmentation. METHODS: A computerized search was conducted on Science Direct, Medline via PubMed, Scopus, and Web of Science using the relevant keywords. English-language studies published between 2013 and 2023 that focused on patients with oral pigmented lesions subjected to different treatment modalities, such as laser or surgical intervention, were compared to determine their efficacy and safety profile. Data were analyzed using R software, applying frequentist models. RESULTS: A total of 27 studies were included. In contrast to the CO2 laser, Er: YAG laser was linked to a higher risk of bleeding (RR = 2.73, p < 0.01), whereas the diode laser had the most favorable score in minimizing bleeding index (P-score = 0.86). In terms of lower risk and postoperative pain score (RR = 0.01, p < 0.01), the Er,Cr:YSGG laser had the most favorable result (P-score = 1.00). The Er: YAG laser demonstrated the highest probability of preventing recurrence (RR = 0.28, p < 0.01), followed by the diode laser (RR = 0.42, p < 0.01). CONCLUSIONS: The choice of treatment for oral pigmentation should be based on individual patient needs and the desired outcomes. The Er: YAG laser seems highly effective in preventing pigment recurrence, the diode laser emerges as a top contender in managing bleeding risks, and the Er,Cr:YSGG laser is particularly efficacious in managing postoperative pain.
ABSTRACT
Licorice (Radix glycyrrhizae) is a plant root extract widely used in various applications, including cosmetics, food supplements, and traditional medicine. It has a long history of medicinal use in different cultures due to its diverse pharmacological properties. Licorice has traditionally been used for treating gastrointestinal problems, respiratory infections, cough, bronchitis, arthritis, and skin conditions. In recent years, the potential therapeutic benefits of licorice for oral health have gained significant interest. This paper aims to provide a comprehensive review of the effects of licorice extracts and their bioactive components on common oral diseases such as dental caries, periodontitis, halitosis, candidiasis, and recurrent aphthous ulcers. The chemical composition of licorice has shown the presence of several bioactive compounds such as glycyrrhizin, glabridin, isoliquiritigenin (ISL), and licochalcone exhibiting various pharmacological activities, including anti-inflammatory, antimicrobial, antioxidative, and immunomodulatory effects. Interestingly, in certain patients, licorice has shown a promising potential to inhibit the spread of viruses, prevent biofilm formation, reduce inflammation, boost immune responses, alleviate pain, and exert antioxidative effects. In this review, we provide a brief overview of the current understanding of licorice's therapeutic benefits in the treatment of oral ailments, emphasising its potential as an alternative treatment option for oral diseases. Further research is warranted to explore its efficacy, safety, and clinical applications using placebo-controlled clinical trials.
ABSTRACT
Abnormal keratinocyte differentiation is fundamental to pathologies such as skin cancer and mucosal inflammatory diseases. The ability to grow keratinocytes in vitro allows the study of differentiation however any translational value is limited if keratinocytes get altered by the culture method. Although serum lipids (SLPs) and phenol red (PR) are ubiquitous components of culture media their effect on differentiation is largely unknown. We show for the first time that PR and SLP themselves suppress expression of differentiation-specific keratins K1, K10 and K2 in normal human epidermal keratinocytes (NHEK) and two important cell lines, HaCaT and N/TERT-1. Removal of SLP increased expression of K1, K10 and K2 in 2D and 3D cultures, which was further enhanced in the absence of PR. The effect was reversed for K1 and K10 by adding all-trans retinoic acid (ATRA) but increased for K2 in the absence of PR. Furthermore, retinoid regulation of differentiation-specific keratins involves post-transcriptional mechanisms as we show KRT2 mRNA is stabilised whilst KRT1 and KRT10 mRNAs are destabilised in the presence of ATRA. Taken together, our results indicate that the presence of PR and SLP in cell culture media may significantly impact in vitro studies of keratinocyte differentiation.
Subject(s)
Cell Differentiation/drug effects , Cell Differentiation/genetics , Gene Expression Regulation/drug effects , Gene Expression Regulation/genetics , Gene Expression/drug effects , Gene Expression/genetics , Keratin-10/genetics , Keratin-10/metabolism , Keratin-1/genetics , Keratin-1/metabolism , Keratin-2/genetics , Keratin-2/metabolism , Keratinocytes/metabolism , Keratinocytes/physiology , Lipids/physiology , Phenolsulfonphthalein/pharmacology , Tretinoin/pharmacology , Cells, Cultured , HaCaT Cells , Humans , RNA, Messenger/genetics , RNA, Messenger/metabolismABSTRACT
Keratin 15 (K15), a type I keratin, which pairs with K5 in epidermis, has been used extensively as a biomarker for stem cells. Two commercial antibodies, LHK15, a mouse monoclonal and EPR1614Y, a rabbit monoclonal, have been widely employed to study K15 expression. Here we report differential reactivity of these antibodies on epithelial cells and tissue sections. Although the two antibodies specifically recognised K15 on western blot, they reacted differently on skin sections and cell lines. LHK15 reacted in patches, whereas EPR1614Y reacted homogenously with the basal keratinocytes in skin sections. In cultured cells, LHK15 did not react with K15 deficient NEB-1, KEB-11, MCF-7 and SW13 cells expressing only exogenous K8 and K18 but reacted when these cells were transduced with K15. On the other hand, EPR1614Y reacted with these cells even though they were devoid of K15. Taken together these results suggest that EPR1614Y recognises a conformational epitope on keratin filaments which can be reconstituted by other keratins as well as by K15. In conclusion, this report highlights that all commercially available antibodies may not be equally specific in identifying the K15 positive stem cell.