Comparative proteomic study of dog and human saliva

<div><p>Saliva contains many proteins that have an important role in biological process of the oral cavity and is closely associated with many diseases. Although the dog is a common companion animal, the composition of salivary proteome and its relationship with that of human are unclear. In this study, shotgun proteomics was used to compare the salivary proteomes of 7 Thai village dogs and 7 human subjects. Salivary proteomes revealed 2,532 differentially expressed proteins in dogs and humans, representing various functions including cellular component organization or biogenesis, cellular process, localization, biological regulation, response to stimulus, developmental process, multicellular organismal process, metabolic process, immune system process, apoptosis and biological adhesion. The oral proteomes of dogs and humans were appreciably different. Proteins related to apoptosis processes and biological adhesion were predominated in dog saliva. Drug-target network predictions by STITCH Version 5.0 showed that dog salivary proteins were found to have potential roles in tumorigenesis, anti-inflammation and antimicrobial processes. In addition, proteins related to regeneration and healing processes such as fibroblast growth factor and epidermal growth factor were also up-regulated in dogs. These findings provide new information on dog saliva composition and will be beneficial for the study of dog saliva in diseased and health conditions in the future.</p></div

Uncovering Ubiquitin and Ubiquitin-like Signaling Networks

Microscopic imaging and technolog

Radiosensitivity Enhancement by Eurycomalactone in Lung Adenocarcinoma via G2/M Cell Cycle Arrest Induction and Delay of DNA Double-strand Break Repair

Background: Radiotherapy (RT) is an important treatment for non-small cell lung cancer (NSCLC). However, the low radiation sensitivity of lung cancer cells turns out to be one major obstacle to RT efficacy. Therefore, application of a sensitizer that increases radiation effect without dose escalation will be beneficial for NSCLC treatment. Eurycoma longifolia Jack is an herbal medicinal plant of South-East Asian origin. The anticancer activity of this plant has also been proved scientifically. Interestingly, among all active quassinoids isolatedfrom E. longifolia, eurycomalactone (ECL) exhibited most potent cytotoxicity against lung cancer cells. Aim: This study aimed to investigate whether ECL would enhance the radiosensitivity of NSCLC cells. Methods: Cytotoxicity of ECL on NSCLC cell lines (A549, Calu-1) and human lung fibroblast (WI-38) wasdetermined using MTT assay. The effects of ECL combined with X-rays on clonogenicity, cell cycle progression, cell apoptosis and the G2/M regulatory proteins expression were analyzed with clonogenic assay, flow cytometry and immunoblotting, respectively. DNA double-strand break (DSB) and the repaircapacity were evaluated by immunofluorescence and immunoblotting.Results/Conclusions: ECL exhibited selective cytotoxicity against A549 cells as compared to the normal one. ECL treatment prior to irradiation synergistically decreased the A549 colony formation, suggesting the enhancement of radiosensitivity by ECL. Moreover, ECL was able to reduce expression of CDK1/2 and CyclinB1 leading to cell cycle arrest at radiosensitive G2/M phase. ECL markedly delayed the repair of radiation-induced DSB. Pre-treatment with ECL followed by X-ray not only delayed the γ-H2AX foci resolving and blocked the 53BP1 foci formation at the DSB sites, but also attenuated expression of DNA repair proteins, Ku80 and KDM4D. Consequently, these effects led to an increase in the cell apoptosis after irradiation. This study offers a great potential for ECL as an alternative safer radiosensitizer to improve the RT efficacy of NSCLC.16th International Congress of Radiation Researc

Enhancement of Radiosensitivity by Eurycomalactone in Human Lung Adenocarcinoma Cells through G2/M Cell Cycle Arrest and Delayed DNA Double-strand Break Repair

Radiotherapy (RT) is an important treatment for non-small cell lung cancer (NSCLC). However, the major obstacles to successful RT includes the low radiosensitivity of cancer cells and the restricted radiation dose which is given without damaging normal tissues. Therefore, the sensitizer that increases RT efficacy without dose escalation will be beneficial for NSCLC treatment. Eurycomalactone (ECL), an active quassinoid isolated from Eurycoma longifolia Jack, has been demonstrated to possess anticancer activity. In this study, we aimed to investigate the effect of ECL on sensitizing NSCLC cells to X-radiation (X-ray) as well as the underlying mechanisms. The results showed that ECL exhibited selective cytotoxicity against the NSCLC cells; A549 and COR-L23 as compared to the normal lung fibroblast. Clonogenic survival results indicated that ECL treatment prior to irradiation synergistically decreased the A549 and COR-L23 colony number. ECL treatment reduced the expression of CyclinB1 and CDK1/2 leading to induce cell cycle arrest at the radiosensitive G2/M phase. Moreover, ECL markedly delayed the repair of radiation-induced DNA double strand breaks (DSB). In A549 cells, pre-treatment with ECL not only delayed the resolving of radiation-induced γ-H2AX foci but also blocked the formation of 53BP1 foci at the DSB sites. In addition, ECL pre-treatment attenuated expression of DNA repair proteins, Ku80 and KDM4D in both NSCLC cells. Consequently, these effects led to an increase of apoptosis in irradiated cells. Thus, ECL radiosensitized the NSCLC cells to X-ray via G2/M arrest induction, delayed the repair of X-ray-induced DSB. This study offers a great potential for ECL as an alternative safer radiosensitizer for increasing the RT efficiency against NSCLC. Materials and methods: Cytotoxicity of ECL was determined using MTT assay on non-small cell lung cancer (NSCLC) cell lines (A549, Calu-1) and human lung fibroblast (WI-38) cells. The effects of ECL combined with X-rays were analyzed using clonogenic assays and that on cell cycle progression and apoptosis by flow cytometry and Western blotting. DNA double-strand breaks (DSB) and DSB repair capacity were evaluated by immunofluorescence and immunoblotting of DSB repair proteins.Results: ECL selectively inhibited the viability of A549 cells but not Calu-1 or WI-38 cells. Pre-treatment with non-cytotoxic (IC20) or cytotoxic (IC50) doses of ECL prior to irradiation synergistically decreased the colony formation rate of cells with the sensitizer enhancement ratios at 10% cell survival of 1.20 and 1.36, respectively. ECL reduced expression of CDK1/2 and cyclinB1, then induced cell cycle arrest at the radiosensitive G2/M transition, leading to increased apoptosis. ECL followed by irradiation delayed the number of γ-H2AX foci, blocked 53BP1 foci formation at the DSB sites, and attenuated expressionof Ku80 and KDM4D. Conclusion: ECL radiosensitized the A549 cell to X-rays via induction of G2/M arrest, delayed X-rays-induced DNA damage repair and enhanced apoptosis. Finally, ECL holds promise as a radiosensitizer for increasing the efficiency of X-ray therapy against lung cancer

Salivary and serum interleukin-17A and interleukin-18 levels in patients with type 2 diabetes mellitus with and without periodontitis.

OBJECTIVE:Interleukin (IL)-17A and IL-18 have been proposed to play important roles in periodontitis and type 2 diabetes mellitus (DM), but human data are conflicting. The present study aimed to investigate the roles of IL-17A and IL-18 in periodontitis and DM by measuring salivary and serum levels, respectively. MATERIALS AND METHODS:A total of 49 participants with type 2 DM and 25 control subjects without type 2 DM were recruited. A periodontal screening and recording (PSR) index (0, 1-2, 3, and 4) was used to classify whether these subjects had periodontitis. Salivary and serum IL-17A and IL-18 levels were measured by enzyme-linked immunosorbent assay. Multiple linear regression analyses were used to evaluate the associations between these cytokines and clinical parameters. RESULTS:Salivary IL-17A levels were not significantly different between patients with DM and controls, however, the levels were significantly higher in controls with periodontitis than those without periodontitis (p = 0.031). Salivary IL-17A levels were significantly associated with the PSR index (β = 0.369, p = 0.011). Multiple linear regression analyses revealed the association of salivary IL-18 levels and fasting plasma glucose (β = 0.270, p = 0.022) whereas serum IL-18 levels were associated with HbA1C (β = 0.293, p = 0.017). No correlation between salivary and serum levels of IL-17A and IL-18 was found. CONCLUSION:Salivary IL-17A was strongly associated with periodontitis, whereas salivary IL-18 was associated with FPG and serum IL-18 was associated with HbA1C. These results suggest the role of these cytokines in periodontal inflammation and DM

Putative salivary protein biomarkers for the diagnosis of oral lichen planus: a case-control study

Abstract Background Salivary protein biomarkers for screening and diagnosis of oral lichen planus (OLP) are not well-defined. The objective of this study was to identify putative protein biomarkers for OLP using proteomic approaches. Methods Pooled unstimulated whole saliva was collected from five OLP patients and five healthy control participants. Saliva samples were then subjected to two-dimensional gel electrophoresis, followed by mass spectrometry to identify putative protein biomarkers. Subsequently, a subset of these putative biomarkers were validated in 24 OLP patients and 24 age-matched healthy control subjects, using an enzyme-linked immunosorbent assay (ELISA). Immunoblotting analyses were then performed in 3 pairs of age- and sex-matched OLP patients and healthy controls to confirm results from the ELISA study. Results Thirty-one protein spots were identified, corresponding to 20 unique proteins. Notably, fibrinogen fragment D and complement component C3c exhibited increased expression in OLP patients, while cystatin SA exhibited decreased expression in OLP patients, compared with healthy control subjects. ELISA analyses indicated increased expression of fibrinogen fragment D and complement component C3c, and decreased expression of cystatin SA, in the saliva of OLP patients. Statistical differences in the expression of salivary complement C3c were observed between OLP patients and healthy control subjects. Immunoblotting analyses confirmed the results of our ELISA study. Conclusion Complement C3c, fibrinogen fragment D and cystatin SA may serve as salivary biomarkers for screening and/or diagnosis of OLP