66 results on '"Punyadeera, Chamindie"'
Search Results
2. One-step homogeneous C-reactive protein assay for saliva
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Punyadeera, Chamindie, Dimeski, Goce, Kostner, Karam, Beyerlein, Peter, and Cooper-White, Justin
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C-reactive protein , *BIOLOGICAL assay , *SALIVA , *CARDIOVASCULAR diseases , *RISK assessment , *MEDICAL screening , *CORONARY disease , *SERUM - Abstract
Abstract: Background: Cardiovascular disease is the leading cause of death in the world. Human C-reactive protein (CRP) has been used in the risk assessment of coronary events. Human saliva mirrors the body''s health and well-being and is non-invasive, easy to collect and ideal for third world countries as well as for large patient screening. The aim was to establish a saliva CRP reference range and to demonstrate the clinical utility of salivary CRP levels in assessing the coronary events in a primary health care setting. Methods: We have used a homogeneous bead based assay to detect CRP levels in human saliva. We have developed a rapid 15min (vs 90min), sequential, one-step assay to detect CRP in saliva. Saliva was collected from healthy volunteers (n=55, ages 20–70years) as well as from cardiac patients (n=28, ages 43–86years). Results: The assay incubation time was optimised from 90min to 15min and generated a positive correlation (n=29, range 10–2189pg/mL, r2=0.94; Passing Bablok slope 0.885, Intercept 0, p>0.10), meaning we could decrease the incubation time and produce equivalent results with confidence. The mean CRP level in the saliva of healthy human volunteers was 285pg/mL and in cardiac patients was 1680pg/mL (p<0.01). Analysis of CRP concentrations in paired serum and saliva samples from cardiac patients gave a positive correlation (r2=0.84, p<0.001) and the salivary CRP concentration capable of distinguishing healthy from diseased patients. Conclusions: The results suggest that this minimally invasive, rapid and sensitive assay will be useful in large patient screening studies for risk assessment of coronary events. [Copyright &y& Elsevier]
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- 2011
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3. A biomarker panel to discriminate between systemic inflammatory response syndrome and sepsis and sepsis severity.
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Punyadeera, Chamindie, Schneider, E. Marion, Schaffer, Dave, Hsin-Yun Hsu, Joos, Thomas O., Kriebel, Fabian, Weiss, Manfred, and Verhaegh, Wim F. J.
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BIOMARKERS , *SEPSIS , *BLOOD proteins , *SEPTIC shock , *METALLOPROTEINASES , *DIFFERENTIAL diagnosis , *STATISTICAL correlation , *CYTOKINES , *CHEMOKINES , *DIAGNOSIS - Abstract
Introduction: In this study, we report on initial efforts to discover putative biomarkers for differential diagnosis of a systemic inflammatory response syndrome (SIRS) versus sepsis; and different stages of sepsis. In addition, we also investigated whether there are proteins that can discriminate between patients who survived sepsis from those who did not. Materials and Methods: Our study group consisted of 16 patients, of which 6 died and 10 survived. We daily measured 28 plasma proteins, for the whole stay of the patients in the ICU. Results: We observed that metalloproteinases and sE-selectin play a role in the distinction between SIRS and sepsis, and that IL-1α, IP-10, sTNF-R2 and sFas appear to be indicative for the progression from sepsis to septic shock. A combined measurement of MMP-3, -10, IL-1α, IP-10, sIL-2R, sFas, sTNF-R1, sRAGE, GM-CSF, IL-1β and Eotaxin allows for a good separation of patients that survived from those that died (mortality prediction with a sensitivity of 79% and specificity of 86%). Correlation analysis suggests a novel interaction between IL-1α and IP-10. Conclusion: The marker panel is ready to be verified in a validation study with or without therapeutic intervention. [ABSTRACT FROM AUTHOR]
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- 2010
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4. The potential of hydrogel‐free tumoroids in head and neck squamous cell carcinoma.
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Wong, Michael, Vasani, Sarju, Breik, Omar, Zhang, Xi, Kenny, Lizbeth, and Punyadeera, Chamindie
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EXTRACELLULAR matrix proteins , *LITERATURE reviews , *SQUAMOUS cell carcinoma , *TUMOR microenvironment , *BASAL lamina - Abstract
Background: Head and neck malignancy, and in particular squamous cell carcinoma (SCC), is responsible for a significant disease burden globally. The lack of an optimal in vitro model system to accurately recapitulate in vivo response to therapy in HNSCC remains a challenge. The development of patient‐derived three‐dimensional tumour cultures, or tumoroids, has enabled improved modelling of the tumour microenvironment through simulation of important characteristics such as tumour hypoxia, cell–cell interactions and nutrient diffusion characteristics. Methods: We performed a comprehensive English‐language literature review of current methods of tumoroid development utilising Matrigel and Cultrex Basement Membrane Extract 2 (key terms: tumour organoids, tumoroids, hydrogels, Matrigel, Cultrex, squamous cell carcinoma, head and neck)—two common proprietary murine‐derived hydrogels containing extracellular matrix proteins. Nascent literature on the establishment of a novel hydrogel‐free platform for tumoroid development as distinct from these existing methods was also explored. Results: Whilst useful for facilitating cell‐matrix interactions and providing a scaffold for three‐dimensional cell growth and organisation, murine‐derived cell matrix methods were noted to have notable limitations including temperature sensitivity and the medium forming a barrier to analysis of the supernatant. A novel hydrogel‐free method of establishing in vitro tumoroid cultures has been subject to experimentation in colorectal but not head and neck malignancy. The absence of a hydrogel provides for the de novo synthesis of extracellular matrix native to the tumour and self‐organisation of cells within this scaffold through the use of ultralow attachment plates. This model demonstrates similar structural and physiological properties to native tissue, whilst enabling more accurate biomimicry of the tumour microenvironment for drug testing. Conclusions: In the absence of prior experimentation on a hydrogel‐free method for establishing HNSCC tumoroids, and comparisons between hydrogel and hydrogel‐free models, the future development of a comparative protocol encompassing recruitment, collection, processing and analysis represents a valuable opportunity. [ABSTRACT FROM AUTHOR]
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- 2024
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5. Circulating tumour cells predict recurrences and survival in head and neck squamous cell carcinoma patients.
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Zhang, Xi, Weeramange, Chameera Ekanayake, Hughes, Brett G. M., Vasani, Sarju, Liu, Zhen Yu, Warkiani, Majid, Hartel, Gunter, Ladwa, Rahul, Thiery, Jean Paul, Kenny, Liz, Breik, Omar, and Punyadeera, Chamindie
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SQUAMOUS cell carcinoma , *CANCER relapse , *SURVIVAL rate , *NECK , *TUMORS - Abstract
Patients with head and neck squamous cell carcinoma (HNSCC) are at a high risk of developing recurrence and secondary cancers. This study evaluates the prognostic and surveillance utilities of circulating tumour cells (CTCs) in HNSCC. A total of 154 HNSCC patients were recruited and followed up for 4.5 years. Blood samples were collected at baseline and follow-up. CTCs were isolated using a spiral microfluid device. Recurrence and death due to cancer were assessed during the follow-up period. In patients with HNSCC, the presence of CTCs at baseline was a predictor of recurrence (OR = 8.40, p < 0.0001) and death (OR= ∞, p < 0.0001). Patients with CTCs at baseline had poor survival outcomes (p < 0.0001). Additionally, our study found that patients with CTCs in a follow-up appointment were 2.5 times more likely to experience recurrence or death from HNSCC (p < 0.05) prior to their next clinical visit. Our study highlights the prognostic and monitoring utilities of CTCs' in HNSCC patients. Early identification of CTCs facilitates precise risk assessment, guiding treatment choices and ultimately enhancing patient outcomes. [ABSTRACT FROM AUTHOR]
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- 2024
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6. Electrochemical immunosensor for the quantification of galectin-3 in saliva.
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Pittman, Trey W., Zhang, Xi, Punyadeera, Chamindie, and Henry, Charles S.
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GALECTINS , *SALIVA , *CARBON electrodes , *HEALTH services accessibility , *HEART failure , *DETECTION limit - Abstract
Heart failure (HF) is an emerging epidemic and remains a major clinical and public health problem. Advances in the healthcare management of HF may lead to lower morbidity and mortality rates but require diagnostics to guide the process. Current diagnostics/prognostics approaches rely on expensive equipment, centralized facilities and trained personnel, marginalizing healthcare access in developing countries and rural communities. These issues have led researchers to focus on developing portable and affordable diagnostics that can be deployed at the point-of-care (POC). Typically, HF biomarkers are measured in blood not saliva. Recently, our team correlated concentrations of salivary Galectin-3 (Gal-3) to outcomes in patients with HF. We have developed an analytical device which consists of an immunoassay based on a screen-printed carbon electrode (SPCE) to quantify Gal-3 levels in saliva samples. Using 10 µL of saliva, the proposed electrochemical immunoassay achieved a concentration dependent signal response in the clinically relevant range with a limit of detection of 9.66 ng/mL. In addition, the storage stability of the modified electrode was investigated, and only a 10.9 % loss in current response over a 35-day period. The results of the immunoassay on the modified SPCEs suggest validity as a POC biosensor system for the management of HF. • Developed an immunosensor on a screen-printed carbon electrode for heart failure biomarker. • Electrochemical detection of Galectin-3 in saliva. • Sensitive detection in clinically relevant concentration range with LOD of 9.66 ng/mL • an important first step towards an electrochemical, non-invasive POC quantitative test for Gal-3, a HF biomarker. [ABSTRACT FROM AUTHOR]
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- 2024
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7. Human papillomavirus (HPV) DNA methylation changes in HPV-associated head and neck cancer.
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Weeramange, Chameera Ekanayake, Tang, Kai Dun, Irwin, Darryl, Hartel, Gunter, Langton-Lockton, Julian, Ladwa, Rahul, Kenny, Lizbeth, Taheri, Touraj, Whitfield, Bernard, Vasani, Sarju, and Punyadeera, Chamindie
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HUMAN papillomavirus , *DNA methylation , *HEAD & neck cancer , *DROOLING , *METHYLATION - Abstract
Despite the rising incidence, currently, there are no early detection methods for HPV-driven HNC (HPV-HNC). Cervical cancer studies suggest that HPV DNA methylation changes can be used as a biomarker to discriminate cancer patients from HPV-infected individuals. As such, this study was designed to establish a protocol to evaluate DNA methylation changes in HPV late genes and long control region (LCR) in saliva samples of HPV-HNC patients and HPV-positive controls. Higher methylation levels were detected in HPV late genes (L1 and L2) in both tumour and saliva samples of HPV-HNC patients compared with HPV-positive controls. Moreover, methylation patterns between tumours and corresponding saliva samples were observed to have a strong correlation (Passing-Bablok regression analysis; τ = 0.7483, P < 0.0001). Considering the differences between HNC and controls in methylation levels in late genes, and considering primer amplification efficiencies, 13 CpG sites located at L1 and L2 genes were selected for further evaluation. A total of 18 HNC saliva samples and 10 control saliva samples were assessed for the methylation levels in the selected sites. From the CpG sites evaluated statistically significant differences were identified for CpG sites at L2—CpG 6 (P = 0.0004), L1—CpG 3 (P = 0.0144), L1—CpG 2 (P = 0.0395) and L2—CpG 19 (P = 0.0455). Our pilot data indicate that higher levels of DNA methylation in HPV late genes are indicative of HPV-HNC risk, and it is a potential supplementary biomarker for salivary HPV detection-based HPV-HNC screening. [ABSTRACT FROM AUTHOR]
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- 2024
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8. A pilot study to investigate the feasibility of transporting saliva samples at room temperature with MAWI Cell Stabilization buffer.
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Lim, Yenkai and Punyadeera, Chamindie
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PAROTID glands , *ARTIFICIAL saliva , *EXOCRINE secretions , *SALIVARY glands , *SPITTING (Oral habit) - Abstract
Saliva is considered as the front-line of non-invasive diagnostics as novel biomarkers continue to emerge for an array of systemic diseases. Biomarker development pipeline relies heavily on pre-analytical process such as saliva collection, handling, transport and storage. The aim of this study was to systematically evaluate the applicability of MAWI Cell Stabilization (MCS) buffer to transport and store saliva samples at room temperature for downstream applications. Human and bacterial genomic DNA (gDNA) and total protein in saliva samples with and without MCS buffer were quantified for a week at three time-points at room temperature. Based on our findings, MCS buffer was able to preserve human gDNA and total protein within the testing time-points. While bacterial gDNA was accurately preserved, MCS buffer was unable to halt bacterial growth at room temperature. We have identified a non-alcohol-based, non-lytic buffer that could maintain the integrity of both genomic materials and proteins in saliva samples. MCS buffer offers a method to potentially transport and store saliva samples at room temperature, accelerating the translation of salivary assays in remote/rural and resource limited settings. [ABSTRACT FROM AUTHOR]
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- 2018
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9. Prognostic utility of serum NT-proBNP (fragments 1-76aa and 13-71aa) and galectin-3 in predicting death and re-hospitalisation due to cardiovascular events in patients with heart failure.
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Zhang, Xi, Wan, Yunxia, Karunathilaka, Nuwan, Chan, Wandy, Kostner, Karam, Hartel, Gunter, Coats, Andrew J. S., Atherton, John J., and Punyadeera, Chamindie
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HEART failure patients , *GALECTINS , *SURVIVAL analysis (Biometry) , *PROGNOSIS ,CARDIOVASCULAR disease related mortality - Abstract
Patients with heart failure (HF) are at a higher risk of rehospitalisation. In this study, we investigated the prognostic utility of galectin-3 (Gal-3) and NT-proBNP fragments (1-76aa and 13-71aa) as biomarkers to predict outcomes for patients with HF. We collected blood samples from patients with HF (n = 101). Gal-3 and NT-proBNP fragments (1–76aa and 13–71aa) concentrations were measured by immunoassay. Survival analysis and Cox proportional regression models were used to determine the prognostic utility of Gal-3 and NT-proBNP fragments. In patients with increased baseline levels of NT-proBNP1-76 the time to primary endpoint (cardiovascular death or re-hospitalisation) was significantly shorter (p = 0.0058), but not in patient with increased baseline levels of Gal-3 or NTproBNP13-71. Patients with increased levels of NT-proBNP13-71aa at 1 month showed reduced time to the primary endpoint (p = 0.0123). Our findings demonstrated that Gal-3 and NT-proBNP can be used as prognostic biomarkers to stratify patients with HF. [ABSTRACT FROM AUTHOR]
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- 2024
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10. Human saliva as a tool to investigate intimate partner violence
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Punyadeera, Chamindie
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- 2012
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11. Author's reply.
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Punyadeera, Chamindie, Schneider, E. Marion, Schaffer, Dave, Hsin-Yun Hsu, Joos, Thomas O., Kriebel, Fabian, Weiss, Manfred, and Verhaegh, Wim F. J.
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LETTERS to the editor , *BIOMARKERS - Abstract
A response by Chamindie Punyadeera and colleague to a letter to the editor about his article "A biomarker panel to discriminate between SIRS and sepsis and sepsis severity," in the 2010 issue is presented.
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- 2010
12. Proteome profiling of salivary small extracellular vesicles in glioblastoma patients.
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Müller Bark, Juliana, Trevisan França de Lima, Lucas, Zhang, Xi, Broszczak, Daniel, Leo, Paul J., Jeffree, Rosalind L., Chua, Benjamin, Day, Bryan W., and Punyadeera, Chamindie
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EXTRACELLULAR vesicles , *GLIOBLASTOMA multiforme , *APOPTOSIS , *CELL communication , *PROGNOSIS - Abstract
Background: Extracellular vesicles (EVs) play a critical role in intercellular communication under physiological and pathological conditions, including cancer. EVs cargo reflects their cell of origin, suggesting their utility as biomarkers. EVs are detected in several biofluids, and their ability to cross the blood–brain barrier has highlighted their potential as prognostic and diagnostic biomarkers in gliomas, including glioblastoma (GBM). Studies have demonstrated the potential clinical utility of plasma‐derived EVs in glioma. However, little is known about the clinical utility of saliva‐derived EVs in GBM. Methods: Small EVs were isolated from whole mouth saliva of GBM patients pre‐ and postoperatively. Isolation was performed using differential centrifugation and/or ultracentrifugation. EVs were characterized by concentration, size, morphology, and EVs cell‐surface protein markers. Protein cargo in EVs was profiled using mass spectrometry. Results: There were no statistically significant differences in size and concentration of EVs derived from pre‐ and post GBM patients' saliva samples. A higher number of proteins were detected in preoperative samples compared to postoperative samples. The authors found four highly abundant proteins (aldolase A, 14‐3‐3 protein ε, enoyl CoA hydratase 1, and transmembrane protease serine 11B) in preoperative saliva samples from GBM patients with poor outcomes. Functional enrichment analysis of pre‐ and postoperative saliva samples showed significant enrichment of several pathways, including those related to the immune system, cell cycle and programmed cell death. Conclusions: This study, for the first time, demonstrates the feasibility of isolating and characterizing small EVs from pre‐ and postoperative saliva samples from GBM patients. Preliminary findings encourage further large cohort validation studies on salivary small EVs to evaluate prognosis in GBM. The current study demonstrates the feasibility of isolation and characterization of small extracellular vesicles (EVs) from saliva samples of glioblastoma (GBM) patients. It also demonstrates that the proteome content of EVs revealed GBM‐related proteins, some already described in previous studies using GBM cell‐conditioned media and plasma of GBM patients. Preliminary findings encourage further studies on salivary small EVs as a source of noninvasive prognostic biomarkers to predict outcomes in patients with GBM. [ABSTRACT FROM AUTHOR]
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- 2023
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13. Circulating tumour DNA alterations: emerging biomarker in head and neck squamous cell carcinoma.
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Huang, Xiaomin, Duijf, Pascal H. G., Sriram, Sharath, Perera, Ganganath, Vasani, Sarju, Kenny, Lizbeth, Leo, Paul, and Punyadeera, Chamindie
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SQUAMOUS cell carcinoma , *CIRCULATING tumor DNA , *PROGNOSIS , *HEAD & neck cancer , *ONCOGENIC viruses , *BIOMARKERS - Abstract
Head and Neck cancers (HNC) are a heterogeneous group of upper aero-digestive tract cancer and account for 931,922 new cases and 467,125 deaths worldwide. About 90% of these cancers are of squamous cell origin (HNSCC). HNSCC is associated with excessive tobacco and alcohol consumption and infection with oncogenic viruses. Genotyping tumour tissue to guide clinical decision-making is becoming common practice in modern oncology, but in the management of patients with HNSCC, cytopathology or histopathology of tumour tissue remains the mainstream for diagnosis and treatment planning. Due to tumour heterogeneity and the lack of access to tumour due to its anatomical location, alternative methods to evaluate tumour activities are urgently needed. Liquid biopsy approaches can overcome issues such as tumour heterogeneity, which is associated with the analysis of small tissue biopsy. In addition, liquid biopsy offers repeat biopsy sampling, even for patients with tumours with access limitations. Liquid biopsy refers to biomarkers found in body fluids, traditionally blood, that can be sampled to provide clinically valuable information on both the patient and their underlying malignancy. To date, the majority of liquid biopsy research has focused on blood-based biomarkers, such as circulating tumour DNA (ctDNA), circulating tumour cells (CTCs), and circulating microRNA. In this review, we will focus on ctDNA as a biomarker in HNSCC because of its robustness, its presence in many body fluids, adaptability to existing clinical laboratory-based technology platforms, and ease of collection and transportation. We will discuss mechanisms of ctDNA release into circulation, technological advances in the analysis of ctDNA, ctDNA as a biomarker in HNSCC management, and some of the challenges associated with translating ctDNA into clinical and future perspectives. ctDNA provides a minimally invasive method for HNSCC prognosis and disease surveillance and will pave the way in the future for personalized medicine, thereby significantly improving outcomes and reducing healthcare costs. [ABSTRACT FROM AUTHOR]
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- 2023
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14. Method optimisation to enrich small extracellular vesicles from saliva samples.
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Jangholi, Abolfazl, Bark, Juliana Müller, Trevisan França de Lima, Lucas, Lima, Luize Goncalves, Möller, Andreas, Kenny, Lizbeth, Vasani, Sarju, Rao, Sudha, Dolcetti, Riccardo, and Punyadeera, Chamindie
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EXTRACELLULAR vesicles , *SALIVA , *GEL permeation chromatography - Abstract
Venn diagram comparing common and unique proteins identified in sEV samples derived from saliva (A) and plasma (B) isolated by UC, UCF, SEC and DG. In saliva sEVs, UC (63) and DG (54) showed a higher representation of the top 100 EV proteins when compared to UCF (45) and SEC (53). Seven micrograms of protein for each isolation method using saliva and plasma samples (N1, N2 and N3) was separated and then blotted in the presence of different antibodies. In contrast, for plasma-derived sEVs, UC, UCF and DG displayed 22, 21 and 20 common proteins, respectively, when compared to the top 100 EV proteins, whilst SEC showed 52 common proteins. [Extracted from the article]
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- 2023
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15. Salivary exosomes as potential biomarkers in cancer.
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Nair, Soumyalekshmi, Tang, Kai Dun, Punyadeera, Chamindie, and Kenny, Liz
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EXOSOMES , *CANCER , *METASTASIS , *SALIVA , *CELL membranes , *ASCITIC fluids - Abstract
Over the past decade, there has been emerging research in the field of extracellular vesicles, especially those originating from endosomes, referred to as 'exosomes. Exosomes are membrane-bound nanovesicles secreted by most cell types upon fusion of multivesicular bodies (MVBs) to the cell plasma membrane. These vesicles are present in almost all body fluids such as blood, urine, saliva, breast milk, cerebrospinal and peritoneal fluids. Exosomes participate in intercellular communication by transferring the biologically active molecules like proteins, nucleic acids, and lipids to neighboring cells. Exosomes are enriched in the tumour microenvironment and growing evidence demonstrates that exosomes mediate cancer progression and metastasis. Given the important biological role played by these nanovesicles in cancer pathogenesis, these can be used as ideal non-invasive biomarkers in detecting and monitoring tumours as well as therapeutic targets. The scope of the current review is to provide an overview of exosomes with a special focus on salivary exosomes as potential biomarkers in head and neck cancers. [ABSTRACT FROM AUTHOR]
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- 2018
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16. Salivary micro RNAs as biomarkers for oropharyngeal cancer.
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Ekanayake Weeramange, Chameera, Tang, Kai Dun, Barrero, Roberto A., Hartel, Gunter, Liu, Zhen, Ladwa, Rahul, Langton‐Lockton, Julian, Frazer, Ian, Kenny, Lizbeth, Vasani, Sarju, and Punyadeera, Chamindie
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MICRORNA , *OROPHARYNGEAL cancer , *HEAD & neck cancer , *TUMOR markers , *HUMAN papillomavirus , *NON-coding RNA - Abstract
Background: Despite the rising incidence, particularly of the human papillomavirus (HPV)‐associated fraction of oropharyngeal cancer (OPC), there are no early detection methods for OPC. Considering the close association between saliva and head and neck cancers, this study was designed to investigate salivary micro RNA (miRNAs) associated with OPC, especially focusing on HPV‐positive OPC. Methods: Saliva was collected from OPC patients at diagnosis and patients were clinically followed up ≤5 years. Salivary small RNA isolated from HPV‐positive OPC patients (N = 6), and HPV‐positive (N = 4) and negative controls (N = 6) were analysed by next‐generation sequencing to identify dysregulated miRNAs. Discovered miRNAs were validated by quantitative PCR using two different assays in a separate cohort of patients (OPC = 91, controls = 92). The relative expression was calculated considering SNORD‐96A as the normalizer. Candidate miRNAs with diagnostic and prognostic potential were evaluated by generalized logistic regression. Results: A panel consisting of nine miRNAs was identified to have the best diagnostic performance to discriminate HPV‐positive OPC from HPV‐positive controls (AUC‐ validation‐1 = 94.8%, validation‐2 = 98%). Further, a panel consisting of six miRNAs were identified to discriminate OPC from controls regardless of the HPV status (AUC‐ validation‐1 = 77.2%, validation‐2 = 86.7%). In addition, the downregulation of hsa‐miR‐7‐5p was significantly associated with poor overall survival of OPC patients (HR = 0.638). A panel consisting of nine miRNAs were identified for the prediction of the overall survival of the OPC patients (log‐rank test‐p = 0.0008). Conclusion: This study highlights that salivary miRNAs can play an essential role in the detection and prognostication of OPC. [ABSTRACT FROM AUTHOR]
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- 2023
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17. A pilot study: Metabolic profiling of plasma and saliva samples from newly diagnosed glioblastoma patients.
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Bark, Juliana Muller, Karpe, Avinash V., Doecke, James D., Leo, Paul, Jeffree, L., Chua, Benjamin, Day, Bryan W., Beale, David J., and Punyadeera, Chamindie
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GLIOBLASTOMA multiforme , *MULTIVARIATE analysis , *SALIVA , *PROGNOSIS , *SALIVA analysis - Abstract
Background: Despite aggressive treatment, more than 90% of glioblastoma (GBM) patients experience recurrences. GBM response to therapy is currently assessed by imaging techniques and tissue biopsy. However, difficulties with these methods may cause misinterpretation of treatment outcomes. Currently, no validated therapy response biomarkers are available for monitoring GBM progression. Metabolomics holds potential as a complementary tool to improve the interpretation of therapy responses to help in clinical interventions for GBM patients. Methods: Saliva and blood from GBM patients were collected pre and postoperatively. Patients were stratified conforming their progression-free survival (PFS) into favourable or unfavourable clinical outcomes (>9 months or PFS = 9 months, respectively). Analysis of saliva (whole-mouth and oral rinse) and plasma samples was conducted utilising LC-QqQ-MS and LC-QTOF-MS to determine the metabolomic and lipidomic profiles. The data were investigated using univariate and multivariate statistical analyses and graphical LASSO-based graphic network analyses. Results: Altogether, 151 metabolites and 197 lipids were detected within all saliva and plasma samples. Among the patients with unfavourable outcomes, metabolites such as cyclic-AMP, 3-hydroxy-kynurenine, dihydroorotate, UDP and cis-aconitate were elevated, compared to patients with favourable outcomes during pre-and post-surgery. These metabolites showed to impact the pentose phosphate and Warburg effect pathways. The lipid profile of patients who experienced unfavourable outcomes revealed a higher heterogeneity in the abundance of lipids and fewer associations between markers in contrast to the favourable outcome group. Conclusion: Our findings indicate that changes in salivary and plasma metabolites in GBM patients can potentially be employed as less invasive prognostic biomarkers/biomarker panel but validation with larger cohorts is required. [ABSTRACT FROM AUTHOR]
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- 2023
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18. Head and neck cancer patient-derived tumouroid cultures: opportunities and challenges.
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Basnayake, B. W. M. Thilini J., Leo, Paul, Rao, Sudha, Vasani, Sarju, Kenny, Lizbeth, Haass, Nikolas K., and Punyadeera, Chamindie
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Head and neck cancers (HNC) are the seventh most prevalent cancer type globally. Despite their common categorisation, HNCs are a heterogeneous group of malignancies arising in various anatomical sites within the head and neck region. These cancers exhibit different clinical and biological manifestations, and this heterogeneity also contributes to the high rates of treatment failure and mortality. To evaluate patients who will respond to a particular treatment, there is a need to develop in vitro model systems that replicate in vivo tumour status. Among the methods developed, patient-derived cancer organoids, also known as tumouroids, recapitulate in vivo tumour characteristics including tumour architecture. Tumouroids have been used for general disease modelling and genetic instability studies in pan-cancer research. However, a limited number of studies have thus far been conducted using tumouroid-based drug screening. Studies have concluded that tumouroids can play an essential role in bringing precision medicine for highly heterogenous cancer types such as HNC. [ABSTRACT FROM AUTHOR]
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- 2023
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19. Head and neck cancer N-glycome traits are cell line and HPV status–dependent.
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Rasheduzzaman, Mohammad, Murugan, Abarna V. M., Zhang, Xi, Oliveira, Tiago, Dolcetti, Riccardo, Kenny, Liz, Johnson, Newell W., Kolarich, Daniel, and Punyadeera, Chamindie
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OROPHARYNX , *HEAD & neck cancer , *ELECTROSPRAY ionization mass spectrometry , *CELL lines , *POST-translational modification , *TANDEM mass spectrometry , *LYSOSOMES ,KERATINOCYTE differentiation - Abstract
Glycosylation is the most common post-translational modification of proteins, and glycosylation changes at cell surfaces are frequently associated with malignant epithelia including head and neck squamous cell carcinoma (HNSCC). In HNSCC, 5-year survival remains poor, averaging around 50% globally: this is partly related to late diagnosis. Specific protein glycosylation signatures on malignant keratinocytes have promise as diagnostic and prognostic biomarkers and as therapeutic targets. Nevertheless, HNSCC-specific glycome is to date largely unknown. Herein, we tested six established HNSCC cell lines to capture the qualitative and semi-quantitative N-glycome using porous graphitized carbon liquid chromatography coupled to electrospray ionisation tandem mass spectrometry. Oligomannose-type N-glycans were the predominant features in all HNSCC cell lines analysed (57.5–70%). The levels of sialylated N-glycans showed considerable cell line-dependent differences ranging from 24 to 35%. Importantly, α2-6 linked sialylated N-glycans were dominant across most HNSCC cell lines except in SCC-9 cells where similar levels of α2-6 and α2-3 sialylated N-glycans were observed. Furthermore, we found that HPV-positive cell lines contained higher levels of phosphorylated oligomannose N-glycans, which hint towards an upregulation of lysosomal pathways. Almost all fucose-type N-glycans carried core-fucose residues with just minor levels (< 4%) of Lewis-type fucosylation identified. We also observed paucimannose-type N-glycans (2–5.5%), though in low levels. Finally, we identified oligomannose N-glycans carrying core-fucose residues and confirmed their structure by tandem mass spectrometry. This first systematic mapping of the N-glycome revealed diverse and specific glycosylation features in HNSCC, paving the way for further studies aimed at assessing their possible diagnostic relevance. [ABSTRACT FROM AUTHOR]
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- 2022
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20. Modelling reoxygenation effects in non-small cell lung cancer cell lines and showing epithelial-mesenchymal transition.
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Kapeleris, Joanna, Müller Bark, Juliana, Ranjit, Shanon, Richard, Derek, Vela, Ian, O'Byrne, Kenneth, and Punyadeera, Chamindie
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NON-small-cell lung carcinoma , *EPITHELIAL-mesenchymal transition , *CELL lines , *CANCER cells - Abstract
Purpose: Circulating tumour cells (CTCs) are a rare cell subpopulation regulated by the tumour microenvironment. In hypoxic conditions, CTCs are able to invade the lymphatic and circulatory systems leading to metastasis at distant sites. Methods: To mimic in vivo oxygen variations and effects on CTCs, we have cultured five non-small cell lung cancer (NSCLC) cell lines under normoxic and hypoxic conditions, followed by a pulse of reoxygenation for 4 h. Results: Proliferation, spheroid-formation and colony formation ability under varying O2 levels were investigated. Proliferation rate was not altered when cells were cultured in 2D models under hypoxic conditions. However, we observed that hypoxia enhanced in vitro formation of tumour-spheres and accelerated clonogenicity of NSCLC cell lines. In addition, cells exposed to hypoxia and reoxygenation conditions showed altered expression of epithelial-mesenchymal transition (EMT) related genes in NSCLC cell lines both at mRNA (AKT1, CAMK2NH1, DESI1, VIM, MAP1B, EGFR, ZEB1, HIF1α) and protein levels (Vimentin, Pan-cytokeratin). Conclusion: Our data suggest that when investigating CTCs as a prognostic biomarker in NSCLC, it is also essential to take into consideration EMT status to obtain a comprehensive overview of CTCs in circulation. [ABSTRACT FROM AUTHOR]
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- 2022
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21. Variation of Human Salivary O-Glycome.
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Kozak, Radoslaw P., Urbanowicz, Paulina A., Punyadeera, Chamindie, Reiding, Karli R., Jansen, Bas C., Royle, Louise, Spencer, Daniel I., Fernandes, Daryl L., and Wuhrer, Manfred
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SALIVA analysis , *GLYCOSYLATION , *BIOMARKERS , *GLYCANS , *ELECTROSPRAY ionization mass spectrometry - Abstract
The study of saliva O-glycosylation is receiving increasing attention due to the potential of glycans for disease biomarkers, but also due to easy access and non-invasive collection of saliva as biological fluid. Saliva is rich in glycoproteins which are secreted from the bloodstream or produced by salivary glands. Mucins, which are highly O-glycosylated proteins, are particularly abundant in human saliva. Their glycosylation is associated with blood group and secretor status, and represents a reservoir of potential disease biomarkers. This study aims to analyse and compare O-glycans released from whole human mouth saliva collected 3 times a day from a healthy individual over a 5 days period. O-linked glycans were released by hydrazinolysis, labelled with procainamide and analysed by ultra-high performance liquid chromatography with fluorescence detection (UHPLC-FLR) coupled to electrospray ionization mass spectrometry (ESI-MS/MS). The sample preparation method showed excellent reproducibility and can therefore be used for biomarker discovery. Our data demonstrates that the O-glycosylation in human saliva changes significantly during the day. These changes may be related to changes in the salivary concentrations of specific proteins. [ABSTRACT FROM AUTHOR]
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- 2016
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22. Application of circulating tumour cells to predict response to treatment in head and neck cancer.
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Zhang, Xi, Ekanayake Weeramange, Chameera, Hughes, Brett G. M., Vasani, Sarju, Liu, Zhen Yu, Warkiani, Majid Ebrahimi, Hartel, Gunter, Ladwa, Rahul, Thiery, Jean Paul, Kenny, Liz, and Punyadeera, Chamindie
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HEAD & neck cancer , *P16 gene , *PATIENT selection , *CANCER relapse , *MICROFLUIDIC devices , *POSITRON emission tomography computed tomography - Abstract
Background: Local recurrence and metastasis remain the major causes of death in head and neck cancer (HNC) patients. Circulating tumour cells (CTCs) are shed from primary and metastatic sites into the circulation system and have been reported to play critical roles in the metastasis and recurrence of HNC. Here, we explored the use of CTCs to predict the response to treatment and disease progression in HNC patients. Methods: Blood samples were collected at diagnosis from HNC patients (n = 119). CTCs were isolated using a spiral microfluidic device and were identified using immunofluorescence staining. Correlation of baseline CTC numbers to 13-week PET-CT data and multidisciplinary team consensus data were conducted. Results: CTCs were detected in 60/119 (50.4%) of treatment naïve HNC patients at diagnosis. Baseline CTC numbers were higher in stage III vs. stage I-II p16-positive oropharyngeal cancers (OPCs) and other HNCs (p = 0.0143 and 0.032, respectively). In addition, we found that baseline CTC numbers may serve as independent predictors of treatment response, even after adjusting for other conventional prognostic factors. CTCs were detected in 10 out of 11 patients exhibiting incomplete treatment responses. Conclusions: We found that baseline CTC numbers are correlated with treatment response in patients with HNC. The expression level of cell-surface vimentin (CSV) on CTCs was significantly higher in patients with persistent or progressive disease, thus providing additional prognostic information for stratifying the risk at diagnosis in HNC patients. The ability to detect CTCs at diagnosis allows more accurate risk stratification, which in the future may be translated into better patient selection for treatment intensification and/or de-intensification strategies. [ABSTRACT FROM AUTHOR]
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- 2022
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23. Analysis of human leukocyte antigen associations in human papillomavirus–positive and –negative head and neck cancer: Comparison with cervical cancer.
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Ekanayake Weeramange, Chameera, Shu, Danhua, Tang, Kai Dun, Batra, Jyotsna, Ladwa, Rahul, Kenny, Lizbeth, Vasani, Sarju, Frazer, Ian H., Dolcetti, Riccardo, Ellis, Jonathan J., Sturm, Richard A., Leo, Paul, and Punyadeera, Chamindie
- Abstract
Background: Although the majority of human papillomavirus (HPV) infections are cleared by the immune system, a small percentage of them progress to develop HPV‐driven cancers. Cervical cancer studies highlight that HPV persistence and cancer risk are associated with genetic factors, especially at the human leukocyte antigen (HLA) genes. This study was conducted to investigate such associations in head and neck cancer (HNC). Methods: In all, 192 patients with HNC and 384 controls were genotyped with the Infinium Global Screening Array (Illumina, Inc). HLA variants were imputed with SNP2HLA, and an association analysis was performed by logistic regression. Results: HPV‐positive HNCs were significantly associated with single‐nucleotide polymorphisms (SNPs) at DRB1_32660090 (P = 1.728 × 10–6) and DRB1_32660116 (P = 1.728 × 10–6) and with the amino acid variant DRB1_11_32660115 (P = 1.728 × 10–6). None of these associations were observed in the HPV‐negative cohort, and this suggested their specificity to convey risk for HPV‐associated HNCs. In general, associations observed for HPV‐negative HNC were relatively weak, and variants in the HLA‐DPA1 region were the strongest among them (P = 4.531 × 10–4). Several lead signals reported by previous HNC genome‐wide association studies, including SNPs rs3135001 (P =.012), rs1049055 (P =.012), and rs34518860 (P =.029) and allele HLA‐DQB1*06 (P =.009), were replicated in the current study. However, these associations were limited to the HPV‐positive HNC group. Several cervical cancer–associated HLA variants, including SNPs rs9272143 (P =.002) and rs9271858 (P =.002) and alleles HLA‐B‐1501 (P =.009) and HLA‐B‐15 (P =.015), were also exclusively associated with HPV‐positive HNC. Conclusions: HPV‐positive HNC risk is associated with distinct HLA variants, and some of them are shared by both cervical cancer and HPV‐positive HNC. Human papillomavirus (HPV)–positive head and neck cancer (HNC) risk is associated with distinct human leukocyte antigen variants, and some of them are shared by both cervical cancer and HPV‐positive HNC. Lay Summary: Cervical cancer studies highlight that human papillomavirus (HPV)–driven cancer risk is linked with human leukocyte antigen (HLA) polymorphism.Hence, the current study was designed to investigate the HLA associations in HPV‐positive and HPV‐negative head and neck cancer (HNC) and compare these associations with cervical cancer.Several lead signals reported by previous HNC and cervical genome‐wide association studies were replicated in the current study.However, these associations were limited to the HPV‐positive HNC group, and this suggests that HPV‐positive HNC risk is associated with distinct HLA variants, and some of them are shared by both cervical cancer and HPV‐positive HNC. [ABSTRACT FROM AUTHOR]
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- 2022
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24. Validation of an immunoassay to measure plasminogen-activator inhibitor-1 concentrations in human saliva.
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Xi Zhang, Dimeski, Goce, and Punyadeera, Chamindie
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EXOCRINE secretions , *FIBRINOLYTIC agents , *IMMUNOGLOBULINS , *CLINICAL chemistry , *IMMUNOLOGY , *SALIVA analysis - Abstract
Introduction: We have previously shown that the concentrations of D-dimer are significantly elevated in saliva compared with plasma. Saliva offers several advantages compared with blood analysis. We hypothesised that human saliva contains plasminogen activator inhibitor-1 (PAI-1) and that the concentrations are not affected by the time of saliva collection. The aim was to adopt and validate an immunoassay to quantify PAI-1 concentrations in saliva and to determine whether saliva collection time has an influence in the measurement. Materials and methods: Two saliva samples (morning and afternoon) from the same day were collected from healthy subjects (N = 40) who have had no underlying heart conditions. A customized AlphaLISA® immunoassay (PerkinElmer® MA, USA) was adopted and used to quantify PAI-1 concentrations. We validated the analytical performance of the customized immunoassay by calculating recovery of known amount of analyte spiked in saliva. Results: The recovery (95.03%), intra- (8.59%) and inter-assay (7.52%) variations were within the acceptable ranges. The median salivary PAI-1 concentrations were 394 pg/mL (interquartile ranges (IQR) 243.4-833.1 pg/mL) in the morning and 376 (129.1-615.4) pg/mL in the afternoon and the plasma concentration was 59,000 (24,000-110,000) pg/mL. Salivary PAI-1 did not correlate with plasma (P = 0.812). Conclusions: The adopted immunoassay produced acceptable assay sensitivity and specificity. The data demonstrated that saliva contains PAI-1 and that its concentration is not affected by the time of saliva collection. There is no correlation between salivary and plasma PAI-1 concentrations. Further studies are required to demonstrate the utility of salivary PAI-1 in CVD risk factor studies. [ABSTRACT FROM AUTHOR]
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- 2014
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25. Identification of salivary N-glycoproteins and measurement of glycosylation site occupancy by boronate glycoprotein enrichment and liquid chromatography/electrospray ionization tandem mass spectrometry.
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Xu, Ying, Bailey, Ulla‐Maja, Punyadeera, Chamindie, and Schulz, Benjamin L.
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GLYCOPROTEINS , *GLYCOSYLATION , *LIQUID chromatography-mass spectrometry , *BIOMARKERS , *TRYPSIN - Abstract
RATIONALE Diseases including cancer and congenital disorders of glycosylation have been associated with changes in the site-specific extent of protein glycosylation. Saliva can be non-invasively sampled and is rich in glycoproteins, giving it the potential to be a useful biofluid for the discovery and detection of disease biomarkers associated with changes in glycosylation. METHODS Saliva was collected from healthy individuals and glycoproteins were enriched using phenylboronic acid based glycoprotein enrichment resin. Proteins were deglycosylated with peptide-N-glycosidase F and digested with AspN or trypsin. Desalted peptides and deglycosylated peptides were separated by reversed-phase liquid chromatography and detected with on-line electrospray ionization quadrupole-time-of-flight mass spectrometry using a 5600 TripleTof instrument. Site-specific glycosylation occupancy was semi-quantitatively determined from the abundance of deglycosylated and nonglycosylated versions of each given peptide. RESULTS Glycoprotein enrichment identified 67 independent glycosylation sites from 24 unique proteins, a 3.9-fold increase in the number of glycosylation sites identified. Enrichment of glycoproteins rather than glycopeptides allowed detection of both deglycosylated and nonglycosylated versions of each peptide, and thereby robust measurement of site-specific occupancy at 21 asparagines. Healthy individuals showed limited biological variability in occupancy, with partially modified sites having characteristics consistent with inefficient glycosylation by oligosaccharyltransferase. Inclusion of negative controls without enzymatic deglycosylation controlled for spontaneous chemical deamidation, and identified asparagines previously incorrectly annotated as glycosylated. CONCLUSIONS We developed a sample preparation and mass spectrometry detection strategy for rapid and efficient measurement of site-specific glycosylation occupancy on diverse salivary glycoproteins suitable for biomarker discovery and detection of changes in glycosylation occupancy in human disease. Copyright © 2014 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]
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- 2014
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26. Paper electrochemical immunosensor for the rapid screening of Galectin-3 patients with heart failure.
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Nihal, Serena, Sarfo, Daniel, Zhang, Xi, Tesfamichael, Tuquabo, Karunathilaka, Nuwan, Punyadeera, Chamindie, and Izake, Emad L.
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GALECTINS , *HEART failure patients , *MEDICAL screening , *FLOW sensors , *CARRIER proteins , *DNA nanotechnology , *HEART assist devices - Abstract
A paper electrochemical immunosensor for the combined binding and quantification of the heart failure (HF) biomarker Galectin-3 has been developed. The simple design of the new sensor is comprised of paper material that is decorated with gold nanostructures, to maximize its electroactive surface area, and functionalized with target-specific recognition molecules to selectively bind the protein from aqueous solutions. The binding of the protein caused the blockage of the electron flow to the sensor electroactive surface, thus causing its oxidation potential to shift and the corresponding current to reduce quantitatively with the increase in the protein concentration within the working range of 0.5ng/mL-8ng/mL (LOQ-0.5 ng/mL). This novel sensor was able to quantify Galectin-3 concentration in saliva samples from HF patients and healthy controls within 20 min with good reproducibility (RSD = 3.64%), without the need for complex sample processing steps. The electrochemical measurements of the patient samples were cross validated by ELISA where the percent agreement between the two methods was found to be 92.7% (RSD = 7.20%). Therefore, the new paper immunosensor sensor has a strong potential for rapid and cost-effective screening of the Galectin 3 biomarker at points of care, thus supporting the timely diagnosis of heart failure. [Display omitted] • A paper electrochemical immunosensor for Galectin-3. • Non-invasive and early and rapid detection of heart damage using human saliva. • The electrochemical measurements agreed with ELISA measurements. • The has strong potential for the detection of the biomarker at points of care. [ABSTRACT FROM AUTHOR]
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- 2024
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27. The deterioration of calcified cartilage integrity reflects the severity of osteoarthritis—A structural, molecular, and biochemical analysis.
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Fan, Xiwei, Wu, Xiaoxin, Trevisan Franca De Lima, Lucas, Stehbens, Samantha, Punyadeera, Chamindie, Webb, Richard, Hamilton, Brett, Ayyapann, Vijay, McLauchlan, Connor, Crawford, Ross, Zheng, Minghao, Xiao, Yin, and Prasadam, Indira
- Abstract
The calcified cartilage zone (CCZ) is a thin interlayer between the hyaline articular cartilage and the subchondral bone and plays an important role in maintaining the joint homeostasis by providing biological and mechanical support from unmineralized cartilage to the underlying mineralized subchondral bone. The hallmark of CCZ characteristics in osteoarthritis (OA) is less well known. The aim of our study is to evaluate the structural, molecular, and biochemical composition of CCZ in tissues affected by primary knee OA and its relationship with disease severity. We collected osteochondral tissue samples stratified according to disease severity, from 16 knee OA patients who underwent knee replacement surgery. We also used meniscectomy‐induced rat samples to confirm the pathophysiologic changes of human samples. We defined the characteristics of the calcified cartilage layer using a combination of morphological, biochemical, proteomic analyses on laser micro‐dissected tissue. Our results demonstrated that the Calcium/Phosphate ratio is unchanged during the OA progression, but the calcium‐binding protein and cadherin binding protein, as well as carbohydrate metabolism‐related proteins, undergo significant changes. These changes were further accompanied by thinning of the CCZ, loss of collagen and proteoglycan content, the occurrence of the endochondral ossification, neovasculature, loss of the elastic module, loss of the collagen direction, and increase of the tortuosity indicating an altered structural and mechanical properties of the CCZ in OA. In conclusion, our results suggest that the calcified cartilage changes can reflect the disease progression. [ABSTRACT FROM AUTHOR]
- Published
- 2022
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28. Saliva proteome research: current status and future outlook.
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Schulz, Benjamin L., Cooper-White, Justin, and Punyadeera, Chamindie K.
- Abstract
Human saliva harbours proteins of clinical relevance and about 30% of blood proteins are also present in saliva. This highlights that saliva can be used for clinical applications just as urine or blood. However, the translation of salivary biomarker discoveries into clinical settings is hampered by the dynamics and complexity of the salivary proteome. This review focuses on the current status of technological developments and achievements relating to approaches for unravelling the human salivary proteome. We discuss the dynamics of the salivary proteome, as well as the importance of sample preparation and processing techniques and their influence on downstream protein applications; post-translational modifications of salivary proteome and protein: protein interactions. In addition, we describe possible enrichment strategies for discerning post-translational modifications of salivary proteins, the potential utility of selected-reaction-monitoring techniques for biomarker discovery and validation, limitations to proteomics and the biomarker challenge and future perspectives. In summary, we provide recommendations for practical saliva sampling, processing and storage conditions to increase the quality of future studies in an emerging field of saliva clinical proteomics. We propose that the advent of technologies allowing sensitive and high throughput proteome-wide analyses, coupled to well-controlled study design, will allow saliva to enter clinical practice as an alternative to blood-based methods due to its simplistic nature of sampling, non-invasiveness, easy of collection and multiple collections by untrained professionals and cost-effective advantages. [ABSTRACT FROM AUTHOR]
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- 2013
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29. Oral contraceptives prevent the development of endometriosis in the chicken chorioallantoic membrane model
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Nap, Annemiek W., Groothuis, Patrick G., Punyadeera, Chamindie, Klein-Hitpass, Ludger, Kamps, Rik, Delvoux, Bert, and Dunselman, Gerard A.J.
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MUCOUS membranes , *ORAL contraceptives , *GYNECOLOGIC drugs , *PROGESTATIONAL hormones - Abstract
Abstract: Background: Fundamental and genetic differences between women in the endometrium may cause some to develop endometriosis, whereas others do not. Oral contraceptives (OC) may have an effect on the endometrium, rendering the development of endometriosis less likely. Study Design: Endometrium from women using OC (OCE) and menstrual endometrium (ME) from normal cycling women were transplanted onto the chicken chorioallantoic membrane (CAM), and endometriosis-like lesion formation was evaluated. Microarray gene expression profiling was performed to identify differentially expressed genes in the endometrium from these groups. Microarray data were validated by real-time PCR. Results: Less endometriosis-like lesions were formed after transplantation of OCE than after transplantation of ME (p<.05). Most of the differentially expressed genes belong to the TGFβ superfamily. Real-time PCR validation revealed that inhibin βA (INHBA) expression was significantly decreased in OCE as compared to ME. Conclusion: OC use affects the characteristics of endometrium, rendering it less potent to develop into endometriosis. [Copyright &y& Elsevier]
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- 2008
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30. Proteomic Alterations in Salivary Exosomes Derived from Human Papillomavirus-Driven Oropharyngeal Cancer.
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Tang, Kai Dun, Wan, Yunxia, Zhang, Xi, Bozyk, Natalie, Vasani, Sarju, Kenny, Liz, and Punyadeera, Chamindie
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EXOSOMES , *OROPHARYNGEAL cancer , *HEAD & neck cancer , *PHOSPHOGLYCERATE kinase , *PYRUVATE kinase , *DEOXYRIBOZYMES , *WESTERN immunoblotting , *LACTATE dehydrogenase - Abstract
Background: Increasing evidence supports the notion that human papillomavirus (HPV) DNA integration onto the human genome can influence and alter the molecular cargo in the exosomes derived from head and neck cancer cells. However, the molecular cargo of salivary exosomes derived from HPV-driven oropharyngeal cancer (HPV-driven OPC) remains unelucidated. Methods and materials: Salivary exosomes morphology and molecular characterizations were examined using the nanoparticle tracking (NTA), western blot analysis, transmission electron microscopy (TEM) and mass spectrometry analysis. Results: We report that HPV16 DNA was detected (80%) in isolated salivary exosomes of HPV-driven OPC patients. Importantly, we demonstrate elevated protein levels of six main glycolytic enzymes [i.e., aldolase (ALDOA), glyceraldehye-3-phosphate dehydrogenase (GAPDH), lactate dehydrogenase A/B (LDHA and LDHB), phosphoglycerate kinase 1 (PGK1) and pyruvate kinase M1/2 (PKM)] in isolated salivary exosomes of HPV-driven OPC patients, suggesting a novel mechanism underlying the potential role of salivary exosomes in mediating the reciprocal interplay between glucose metabolism and HPV-driven OPC. Conclusion: Our data demonstrate the potential diagnostic value of HPV16 DNA and glycolytic enzymes in salivary exosomes in discriminating healthy controls from HPV-driven OPC patients, thereby opening new avenues in the future for clinical translation studies. [ABSTRACT FROM AUTHOR]
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- 2021
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31. Evaluation of sample preparation methods for label-free quantitative profiling of salivary proteome.
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Zhang, Xi, Sadowski, Pawel, and Punyadeera, Chamindie
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CHEMICAL sample preparation , *SALIVA , *PROTEOMICS , *SAMPLING methods , *BODY fluids , *QUANTITATIVE research , *DEPTH profiling - Abstract
Saliva has become one of the more attractive body fluids for protein biomarker discovery studies because of its ease of collection, non-invasiveness and multiple samples can be collected from an individual at a single time point. With the development of modern data acquisition strategies, such as Sequential Window Acquisition of all Theoretical Mass Spectrometry (SWATH-MS), the quality of saliva sample preparation has become a crucial factor for a successful protein identification and quantification. Several sample preparation methods have been proposed, but there has been no systematic evaluation conducted to date that compared each of these methods. We have therefore, performed an extensive assessment using technical and biological repeats to evaluate the number of protein IDs and repeatability of three most commonly used techniques, in-solution digestion, filter-aided sample preparation (FASP) and in stage-tip (iST) digestion. We discovered that in the case of human saliva sample, FASP provided the highest number of proteins (human and microbial) identifiable from a pool saliva sample, and there were no significant differences in terms of repeatability among the three methods investigated. [ABSTRACT FROM AUTHOR]
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- 2020
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32. Circulating biomarkers in patients with glioblastoma.
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Müller Bark, Juliana, Kulasinghe, Arutha, Chua, Benjamin, Day, Bryan W., and Punyadeera, Chamindie
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Gliomas are the most common tumours of the central nervous system and the most aggressive form is glioblastoma (GBM). Despite advances in treatment, patient survival remains low. GBM diagnosis typically relies on imaging techniques and postoperative pathological diagnosis; however, both procedures have their inherent limitations. Imaging modalities cannot differentiate tumour progression from treatment-related changes that mimic progression, known as pseudoprogression, which might lead to misinterpretation of therapy response and delay clinical interventions. In addition to imaging limitations, tissue biopsies are invasive and most of the time cannot be performed over the course of treatment to evaluate 'real-time' tumour dynamics. In an attempt to address these limitations, liquid biopsies have been proposed in the field. Blood sampling is a minimally invasive procedure for a patient to endure and could provide tumoural information to guide therapy. Tumours shed tumoural content, such as circulating tumour cells, cell-free nucleic acids, proteins and extracellular vesicles, into the circulation, and these biomarkers are reported to cross the blood-brain barrier. The use of liquid biopsies is emerging in the field of GBM. In this review, we aim to summarise the current literature on circulating biomarkers, namely circulating tumour cells, circulating tumour DNA and extracellular vesicles as potential non-invasively sampled biomarkers to manage the treatment of patients with GBM. [ABSTRACT FROM AUTHOR]
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- 2020
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33. Two enemies, one fight: An update of oral cancer in patients with Fanconi anemia.
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Amenábar, José M., Torres‐Pereira, Cassius C., Tang, Kai D., Punyadeera, Chamindie, and Torres-Pereira, Cassius C
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FANCONI'S anemia , *ORAL cancer , *HEAD & neck cancer , *SQUAMOUS cell carcinoma , *CANCER risk factors , *CANCER prognosis - Abstract
Fanconi anemia (FA) is a rare inherited genetic condition that may lead to bone marrow failure, leukemia, and/or solid tumors. It is caused by the loss of function of at least 1 gene of the FA/BRCA pathway, which is necessary for DNA repair. Patients with FA have a 200-fold to 1000-fold risk of developing head and neck cancer, mainly oral squamous cell carcinoma (OSCC), and of doing so at a much younger age than individuals within the general population. Also, patients who have FA with OSCC have poor overall survival rates, reinforcing the necessity to detect OSCC early. The scope of the current review is to provide an update on OSCC in patients with FA. [ABSTRACT FROM AUTHOR]
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- 2019
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34. Relationship between p16 expression and prognosis in different anatomic subsites of OSCC.
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Ni, Yanhong, Zhang, Xiaoxin, Wan, Yunxia, Dun Tang, Kai, Xiao, Yin, Jing, Yue, Song, Yuxian, Huang, Xiaofeng, Punyadeera, Chamindie, and Hu, Qingang
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SQUAMOUS cell carcinoma , *PROGNOSIS - Abstract
BACKGROUND: p16 has often been found to be overexpressed in patients oral squamous cell carcinoma (OSCC), but its prognostic value between anatomic subsites is still unclear. OBJECTIVE: The aim of this study was to investigate the diagnostic and prognostic values of p16 in OSCC originating from tongue, gingiva or buccal mucosa. METHODS: A total of 147 OSCC patients with tumors arising from the tongue, gingiva or buccal mucosa were enrolled in this study. p16 expression was detected using immunohistochemistry (IHC), and the presence of HPV16 was determined by real-time PCR in p16 positive patients. The correlation of p16 expression with the clinical parameters was evaluated. RESULTS: Only one p16 positive patient with a cut off value of 25% and 75% was HPV16 positive. Although overall survival (OS), recurrence free survival (RFS) and metastasis free survival (MFS) had no significant differences between the p16 positive and negative patients, p16 negative patients (cut off value 25%) had more RFS in the buccal mucosa cancer (p = 0.03) than the p16-positive patients. CONCLUSIONS: The prevalence of HPV16 in Chinese OSCC patients was low. p16 overexpression decoupled from HPV infection was not a prognostic marker for OSCC patients except for patients with the buccal mucosa cancer. [ABSTRACT FROM AUTHOR]
- Published
- 2019
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35. Circulating tumour cell PD-L1 test for head and neck cancers.
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Kulasinghe, Arutha, Kenny, Liz, and Punyadeera, Chamindie
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HEAD & neck cancer patients , *SQUAMOUS cell carcinoma , *PROGRAMMED cell death 1 receptors , *CANCER cells , *COMPANION diagnostics , *PATIENTS - Abstract
Immune checkpoint inhibitors have gained traction over the last few years in the treatment of metastatic/recurrent head and neck squamous cell carcinoma (HNSCC) patients. Monoclonal antibodies that block the programmed death 1 (PD-1) receptor and its major ligand, PD-L1, have shown durable responses and low toxicity profiles. There are currently no validated predictive biomarkers to select patients likely to respond to anti-PD-1/PD-L1 therapy to avoid unwanted side effects and to reduce healthcare expenditure. A circulating tumour cell (CTC) PD-L1 assay could be developed as a companion diagnostic tool to potentially predict the efficacy of immune checkpoint blockade treatments. [ABSTRACT FROM AUTHOR]
- Published
- 2017
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36. A comparison between mutational profiles in tumour tissue DNA and circulating tumour DNA in head and neck squamous cell carcinoma – A systematic review.
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Huang, Xiaomin, Leo, Paul, Jones, Lee, Duijf, Pascal H.G., Hartel, Gunter, Kenny, Lizbeth, Vasani, Sarju, and Punyadeera, Chamindie
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NECK , *SQUAMOUS cell carcinoma , *SINGLE nucleotide polymorphisms , *HEAD & neck cancer , *CIRCULATING tumor DNA , *TUMORS - Abstract
Head and neck cancer is the seventh most common malignancy globally. Head and neck squamous cell carcinoma (HNSCC) originates from squamous cells and 90% of HNC are HNSCC. The gold standard for diagnosing HNSCC is tissue biopsy. However, given tumour heterogeneity, biopsies may miss important cancer-associated molecular signatures, and more importantly, after the tumour is excised, there is no means of tracking response to treatment in patients. Captured under liquid biopsy, circulating tumour DNA (ctDNA), may identify in vivo molecular genotypes and complements tumour tissue analysis in cancer management. A systematic search was conducted in PubMed, Embase, Scopus and the Cochran Library between 2012 to early 2023 on ctDNA in HNSCC using publications written in English. We summarise 20 studies that compared mutational profiles between tumour tissue DNA (tDNA) and ctDNA, using a cohort of 631 HNSCC patients and 139 controls. Among these studies, the concordance rates varied greatly and the most mutated and the most concordant gene was TP53, followed by PIK3CA, CDKN2A, NOTCH1 and FAT1. Concordant variants were mainly found in Stage IV tumours, and the mutation type is mostly single nucleotide variants (SNV). We conclude that, as a biomarker for HNSCC, ctDNA demonstrates great promise as it recapitulates tumour genotypes, however additional multi-central trials are needed. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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37. The prognostic significance of circulating tumor cells in head and neck and non‐small‐cell lung cancer.
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Kulasinghe, Arutha, Kapeleris, Joanna, Kimberley, Rebecca, Mattarollo, Stephen R., Thompson, Erik W., Thiery, Jean‐Paul, Kenny, Liz, O'Byrne, Ken, and Punyadeera, Chamindie
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LUNG cancer , *HEAD & neck cancer , *CIRCULATING tumor DNA , *TUMORS - Abstract
Tumor biopsy is the gold standard for the assessment of clinical biomarkers for treatment. However, tumors change dynamically in response to therapy, and there remains a need for a more representative biomarker that can be assayed over the course of treatment. Circulating tumor cells (CTCs) may provide clinically important and comprehensive tumoral information that is predictive of treatment response and outcome. Blood samples were processed for CTCs from 56 patients using the ClearCell FX system. Captured cells were phenotyped for CTC clusters and markers for immunotherapy (PD‐L1) CTC chromosomal architecture (ALK, EGFR). CTCs were isolated in 11/23 (47.8%) of head and neck cancer (HNC) patients and 17/33 (51.5%) of non‐small‐cell lung cancer (NSCLC) patients. CTCs were determined to be PD‐L1‐positive in 6/11 (54.4%) HNC and 11/17 (64.7%) NSCLC cases, respectively. 3D chromosomal DNA FISH for ALK and EGFR molecular targets showed better resolution than in 2D when imaging CTCs. HNC CTC‐positive patients had shorter progression‐free survival (PFS) (hazard ratio[HR]: 4.946; 95% confidence internal[CI]:1.571‐15.57; P = 0.0063), and PD‐L1‐positive CTCs were found to be significantly associated with worse outcome ([HR]:5.159; 95% [CI]:1.011‐26.33; P = 0.0485). In the advanced stage NSCLC patient cohort, PFS was not found to be associated with CTCs prior to therapy ([HR]:2.246; 95% [CI]:0.9565‐5.273; P = 0.0632), nor the presence of PD‐L1 expression ([HR]:1.646; 95% [CI]:0.5128‐5.283; P = 0.4023). This study demonstrated that CTCs are predictive of poorer outcomes in HNC and provides distinct and separate utility for CTCs in HNC and NSCLC, which may be more representative of the disease burden and overall survival than the parameters used to measure them. 3D DNA FISH of circulating tumor cells demonstrates a higher resolution for genomic aberrations compared to 2D imaging modalities. [ABSTRACT FROM AUTHOR]
- Published
- 2018
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38. Detecting salivary host and microbiome RNA signature for aiding diagnosis of oral and throat cancer.
- Author
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Banavar, Guruduth, Ogundijo, Oyetunji, Julian, Cristina, Toma, Ryan, Camacho, Francine, Torres, Pedro J., Hu, Lan, Chandra, Tarun, Piscitello, Andrew, Kenny, Liz, Vasani, Sarju, Batstone, Martin, Dimitrova, Nevenka, Vuyisich, Momchilo, Amar, Salomon, and Punyadeera, Chamindie
- Subjects
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CANCER diagnosis , *MEDICAL care costs , *SQUAMOUS cell carcinoma , *RNA , *ORAL cancer - Abstract
• CancerDetect test detects RNA biomarker for oral cancer and throat cancer. • Test specificity 94 %; oral cancer sensitivity 90 %; throat cancer sensitivity 84.2 % • Non-invasive saliva test could allow early detection in various healthcare settings. • Received breakthrough designation by US FDA. • Microbial and human transcript biomarker discovered via machine learning. Oral squamous cell carcinoma (OSCC) and oropharyngeal squamous cell carcinoma (OPSCC) can go undetected resulting in late detection and poor outcomes. We describe the development and validation of CancerDetect for Oral & Throat cancer™ (CDOT), to detect markers of OSCC and/or OPSCC within a high-risk population. We collected saliva samples from 1,175 individuals who were 50 years or older, or adults with a tobacco use history. 945 of those were used to train a classifier using machine learning methods, resulting in a salivary microbial and human metatranscriptomic signature. The classifier was then independently validated on the 230 remaining samples prospectively collected and unseen by the classifier, consisting of 20 OSCC (all stages), 76 OPSCC (all stages), and 134 negatives (including 14 pre-malignant). On the validation cohort, the specificity of the CDOT test was 94 %, sensitivity was 90 % for participants with OSCC, and 84.2 % for participants with OPSCC. Similar classification results were observed among people in early stage (stages I & II) vs late stage (stages III & IV). CDOT is a non-invasive test that can be easily administered in dentist offices, primary care centres and specialised cancer clinics for early detection of OPSCC and OSCC. This test, having received FDA's breakthrough designation for accelerated review, has the potential to enable early diagnosis, saving lives and significantly reducing healthcare expenditure. [ABSTRACT FROM AUTHOR]
- Published
- 2023
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39. PD-L1 expressing circulating tumour cells in head and neck cancers.
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Kulasinghe, Arutha, Perry, Chris, Kenny, Liz, Warkiani, Majid E., Nelson, Colleen, and Punyadeera, Chamindie
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HEAD & neck cancer treatment , *IMMUNOTHERAPY , *APOPTOSIS , *CIRCULATING tumor DNA , *IMMUNOCYTOCHEMISTRY - Abstract
Background: Blockade of the PD-1/PD-L1 immune checkpoint pathway is emerging as a promising immunotherapeutic approach for the management and treatment of head and neck cancer patients who do not respond to 1st/2nd line therapy. However, as checkpoint inhibitors are cost intensive, identifying patients who would most likely benefit from anti PD-L1 therapy is required. Developing a non-invasive technique would be of major benefit to the patient and to the health care system.Case Presentation: We report the case of a 56 year old man affected by a supraglottic squamous cell carcinoma (SCC). A CT scan showed a 20 mm right jugulodigastric node and suspicious lung lesions. The lung lesion was biopsied and confirmed to be consistent with SCC. The patient was offered palliative chemotherapy. At the time of presentation, a blood sample was taken for circulating tumour cell (CTC) analysis. The dissemination of cancer was confirmed by the detection of CTCs in the peripheral blood of the patient, measured by the CellSearch System (Janssen Diagnostics). Using marker-independent, low-shear spiral microfluidic technology combined with immunocytochemistry, CTC clusters were found in this patient at the same time point, expressing PD-L1.Conclusion: This report highlights the potential use of CTCs to identify patients which might respond to anti PD-L1 therapy. [ABSTRACT FROM AUTHOR]- Published
- 2017
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40. A Pilot Study into the Association between Oral Health Status and Human Papillomavirus--16 Infection.
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Bennett, Nigel, Kai Dun Tang, Yenkai Lim, Punyadeera, Chamindie, Charles Xiaohang Sun, Tran, Peter, Samaranayake, Lakshman, and Frazer, Ian
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PAPILLOMAVIRUS disease diagnosis , *PAPILLOMAVIRUS diseases , *ORAL cancer , *HEAD & neck cancer , *ORAL diseases - Abstract
Background: Over the next 20 years, oropharyngeal cancers (OPC) will represent the majority of head and neck cancers (HNCs) in the United States. It is estimated that human papillomavirus (HPV) may account for as much as 70% to 80% of OPCs in North America and in certain parts of Europe. It is hence crucial to understand the disease risk factors and natural history of oral HPV infections. We hypothesized that poor oral health (by measures such as poor oral hygiene and periodontal disease) leads to a higher degree of oral HPV-16 infections within a patient cohort from a dental school clinic. This study aims to test this hypothesis and gauge possible disease associations before larger scale studies. Subjects and Methods: 223 participants were recruited in this study from the University of Queensland Dental School clinic. Clinical oral health parameters (such as oral hygiene measures and periodontal disease measurements) have been examined and determined by dental professionals. We have collected oral rinse samples from these volunteers. Results: 10 (4.5%) out of 223 participants were found to have HPV-16 DNA in their oral rinse samples using NB2 endpoint PCR and Sanger sequencing. Within the HPV-16 DNA positive subjects, 7 (70%) and 3 (30%) were associated with poor oral hygiene and periodontal disease, respectively. Conclusion: Our results show a trend towards a positive correlation between oral HPV-16 infection and poor clinical oral health status. [ABSTRACT FROM AUTHOR]
- Published
- 2017
- Full Text
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41. Within-Day Baseline Variation in Salivary Biomarkers in Healthy Men.
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Idris, Firman Prathama, Wan, Yunxia, Zhang, Xi, and Punyadeera, Chamindie
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BIOLOGICAL variation , *BIOMARKERS , *ELECTROLYTES , *DNA methylation , *INDIVIDUALIZED medicine - Abstract
Saliva is an easily accessible sample and offers practical and noninvasive biomarker solutions as an alternative to blood and urine-based diagnostics. Saliva contains a plethora of biomolecules such as nucleic acids, hormones, proteins, and electrolytes. On the other hand, little is known on the extent to which the biomolecules in saliva vary over time within a given person. This baseline information is crucial for future development of robust saliva-based diagnostics. We have collected unstimulated whole mouth saliva from 20 healthy young men at four times during the day, including before and after a meal. We measured the salivary cortisol, testosterone, C-reactive protein (CRP), stability of genomic DNA (gDNA) and DNA methylation levels of APC, P16INK4a, and PCQAP in these samples. We found that the salivary CRP, DNA methylation, and CD44 gDNA levels did not vary significantly across four time points ( p > 0.05) while the salivary cortisol and testosterone levels significantly varied from the morning collection to the afternoon collection ( p < 0.05). Furthermore, salivary cortisol levels were significantly affected by eating ( p < 0.05). Our study offers a within-person baseline temporal assessment of several clinically relevant biomolecules and diagnostics, and suggests that salivary cortisol and testosterone levels vary over time in a given day whereas CRP and DNA methylation of tumor suppressor genes and CD44 amplification are stable throughout the day. Future research and clinical applications of salivary biomarkers and diagnostics should take into consideration their temporal variations. [ABSTRACT FROM AUTHOR]
- Published
- 2017
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42. Potential use of salivary mRNA-based biomarkers to predict oral squamous cell carcinoma.
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Tang, Kai Dun, Weeramange, Chameera Ekanayake, Vider, Jelena, Hartel, Gunter, West, Nicholas P., McMillan, Nigel A.J., Batstone, Martin D., Liu, Zhen, Vasani, Sarju, Kenny, Liz, and Punyadeera, Chamindie
- Published
- 2022
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43. The development of a liquid biopsy for head and neck cancers.
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Schmidt, Henri, Kulasinghe, Arutha, Kenny, Liz, and Punyadeera, Chamindie
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HEAD & neck cancer treatment , *BIOPSY , *HEAD & neck cancer diagnosis , *EXOSOMES , *BLOOD sampling - Abstract
Developing non-invasive diagnostic tools in the field of head and neck oncology has been a challenge. Analysis of circulating tumour derivatives in a patient's blood has been explored in other solid cancers. This includes analysis of circulating tumour DNA, intact circulating tumour cells (CTCs) and exosomes. These circulating tumour derivatives provide avenues of investigation which can be representative of a patient's primary tumour signature and can be assessed from a patient's blood sample. In advanced stage cancer patients, these tumour derivatives are found in higher amounts, attributed to higher cellular turnover (apoptosis, autophagy), lysed CTCs and sloughing from necrotic tumours. Head and neck squamous cell carcinoma (HNSCC) patients often present with advanced disease associated with a poor 5-year survival of <50%. Outside of sophisticated imaging and clinical examination, there is a lack of available biomarkers to measure disease burden, and/or response to therapy. Implementation of a liquid biopsy in HNSCC through serial blood samples has the potential to detect metastatic events earlier, thereby allowing better selection of appropriate treatment choices, predict prognosis in patients with potentially curable disease, monitor systemic therapies and residual disease post-treatment. [ABSTRACT FROM AUTHOR]
- Published
- 2016
- Full Text
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44. Salivary DNA methylation panel to diagnose HPV-positive and HPV-negative head and neck cancers.
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Yenkai Lim, Yunxia Wan, Vagenas, Dimitrios, Ovchinnikov, Dmitry A., Perry, Chris F. L., Davis, Melissa J., and Punyadeera, Chamindie
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- *
DNA methylation , *PAPILLOMAVIRUS disease diagnosis , *HEAD & neck cancer treatment , *SQUAMOUS cell carcinoma , *TUMOR suppressor genes , *DIAGNOSIS - Abstract
Background: Head and neck squamous cell carcinoma (HNSCC) is a heterogeneous group of tumours with a typical 5 year survival rate of <40 %. DNA methylation in tumour-suppressor genes often occurs at an early stage of tumorigenesis, hence DNA methylation can be used as an early tumour biomarker. Saliva is an ideal diagnostic medium to detect early HNSCC tumour activities due to its proximity to tumour site, non-invasiveness and ease of sampling. We test the hypothesis that the surveillance of DNA methylation in five tumour-suppressor genes (RASSF1α, p16INK4a, TIMP3, PCQAP/MED15) will allow us to diagnose HNSCC patients from a normal healthy control group as well as to discriminate between Human Papillomavirus (HPV)-positive and HPV-negative patients. Methods: Methylation-specific PCR (MSP) was used to determine the methylation levels of RASSF1α, p16INK4a, TIMP3 and PCQAP/MED15 in DNA isolated from saliva. Statistical analysis was carried out using non-parametric Mann- Whitney's U-test for individually methylated genes. A logistic regression analysis was carried out to determine the assay sensitivity when combing the five genes. Further, a five-fold cross-validation with a bootstrap procedure was carried out to determine how well the panel will perform in a real clinical scenario. Results: Salivary DNA methylation levels were not affected by age. Salivary DNA methylation levels for RASSF1α, p16INK4a, TIMP3 and PCQAP/MED15 were higher in HPV-negative HNSCC patients (n = 88) compared with a normal healthy control group (n = 122) (sensitivity of 71 % and specificity of 80 %). Conversely, DNA methylation levels for these genes were lower in HPV-positive HNSCC patients (n = 45) compared with a normal healthy control group (sensitivity of 80 % and specificity of 74 %), consistent with the proposed aetiology of HPV-positive HNSCCs. Conclusions: Salivary DNA tumour-suppressor methylation gene panel has the potential to detect early-stage tumours in HPV-negative HNSCC patients. HPV infection was found to deregulate the methylation levels in HPVpositive HNSCC patients. Large-scale double-blinded clinical trials are crucial before this panel can potentially be integrated into a clinical setting. [ABSTRACT FROM AUTHOR]
- Published
- 2016
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45. Enrichment and identification of glycoproteins in human saliva using lectin magnetic bead arrays.
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Caragata, Michael, Shah, Alok K., Schulz, Benjamin L., Hill, Michelle M., and Punyadeera, Chamindie
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GLYCOPROTEINS , *SALIVA analysis , *LECTINS , *MAGNETIC fields , *GLYCOSYLATION , *BIOMARKERS - Abstract
Aberrant glycosylation of proteins is a hallmark of tumorigenesis and could provide diagnostic value in cancer detection. Human saliva is an ideal source of glycoproteins due to the relatively high proportion of glycosylated proteins in the salivary proteome. Moreover, saliva collection is noninvasive and technically straightforward, and the sample collection and storage is relatively easy. Although differential glycosylation of proteins can be indicative of disease states, identification of differential glycosylation from clinical samples is not trivial. To facilitate salivary glycoprotein biomarker discovery, we optimized a method for differential glycoprotein enrichment from human saliva based on lectin magnetic bead arrays (saLeMBA). Selected lectins from distinct reactivity groups were used in the saLeMBA platform to enrich salivary glycoproteins from healthy volunteer saliva. The technical reproducibility of saLeMBA was analyzed with liquid chromatography–tandem mass spectrometry (LC–MS/MS) to identify the glycosylated proteins enriched by each lectin. Our saLeMBA platform enabled robust glycoprotein enrichment in a glycoprotein- and lectin-specific manner consistent with known protein-specific glycan profiles. We demonstrated that saLeMBA is a reliable method to enrich and detect glycoproteins present in human saliva. [ABSTRACT FROM AUTHOR]
- Published
- 2016
- Full Text
- View/download PDF
46. A pilot study to compare the detection of HPV-16 biomarkers in salivary oral rinses with tumour p16(INK4a) expression in head and neck squamous cell carcinoma patients.
- Author
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Chai, Ryan C., Lim, Yenkai, Frazer, Ian H., Yunxia Wan, Perry, Christopher, Jones, Lee, Lambie, Duncan, Punyadeera, Chamindie, and Wan, Yunxia
- Subjects
- *
BIOMARKERS , *HEAD & neck cancer diagnosis , *SQUAMOUS cell carcinoma , *ORAL cancer patients , *PAPILLOMAVIRUSES , *PILOT projects , *PATIENTS - Abstract
Background: Human papilloma virus-16 (HPV-16) infection is a major risk factor for a subset of head and neck squamous cell carcinoma (HNSCC), in particular oropharyngeal squamous cell carcinoma (OPSCC). Current techniques for assessing the HPV-16 status in HNSCC include the detection of HPV-16 DNA and p16(INK4a) expression in tumor tissues. When tumors originate from hidden anatomical sites, this method can be challenging. A non-invasive and cost-effective alternative to biopsy is therefore desirable for HPV-16 detection especially within a community setting to screen at-risk individuals.Methods: The present study compared detection of HPV-16 DNA and RNA in salivary oral rinses with tumor p16(INK4a) status, in 82 HNSCC patients using end-point and quantitative polymerase chain reaction (PCR).Results: Of 42 patients with p16(INK4a)-positive tumours, 39 (sensitivity = 92.9 %, PPV = 100 % and NPV = 93 %) had oral rinse samples with detectable HPV-16 DNA, using end-point and quantitative PCR. No HPV-16 DNA was detected in oral rinse samples from 40 patients with p16(INK4a) negative tumours, yielding a test specificity of 100 %. For patients with p16(INK4a) positive tumours, HPV-16 mRNA was detected using end-point reverse transcription PCR (RT-PCR) in 24/40 (sensitivity = 60 %, PPV = 100 % and NPV = 71 %), and using quantitative RT-PCR in 22/40 (sensitivity = 55 %, PPV = 100 % and NPV = 69 %). No HPV-16 mRNA was detected in oral rinse samples from the p16(INK4a)-negative patients, yielding a specificity of 100 %.Conclusions: We demonstrate that the detection of HPV-16 DNA in salivary oral rinse is indicative of HPV status in HNSCC patients and can potentially be used as a diagnostic tool in addition to the current methods. [ABSTRACT FROM AUTHOR]- Published
- 2016
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- View/download PDF
47. A paper-based optical sensor for the screening of viruses through the cysteine residues of their surface proteins: A proof of concept on the detection of coronavirus infection.
- Author
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Gholami, Mahnaz D., Guppy-Coles, Kristyan, Nihal, Serena, Langguth, Daman, Sonar, Prashant, Ayoko, Godwin A., Punyadeera, Chamindie, and Izake, Emad L.
- Subjects
- *
COVID-19 , *CORONAVIRUSES , *SARS-CoV-2 , *REVERSE transcriptase polymerase chain reaction , *OPTICAL sensors , *MEDICAL screening , *COLOR change sensors - Abstract
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a serious threat to human health. Current methods such as reverse transcription polymerase chain reaction (qRT-PCR) are complex, expensive, and time-consuming. Rapid, and simple screening methods for the detection of SARS-CoV-2 are critically required to fight the current pandemic. In this work we present a proof of concept for, a simple optical sensing method for the screening of SARS-CoV-2 through its spike protein subunit S1. The method utilizes a target-specific extractor chip to bind the protein from the biological specimens. The disulfide bonds of the protein are then reduced into a biothiol with sulfhydryl (SH) groups that react with a blue-colored benzothiazole azo dye-Hg complex (BAN-Hg) and causes the spontaneous change of its blue color to pink which is observable by the naked eye. A linear relationship between the intensity of the pink color and the logarithm of reduced S1 protein concentration was found within the working range 130 ng.mL−1-1.3 pg mL−1. The lowest limit of detection (LOD) of the assay was 130 fg mL−1. A paper based optical sensor was fabricated by loading the BAN-Hg sensor onto filter paper and used to screen the S1 protein in spiked saliva and patients' nasopharyngeal swabs. The results obtained by the paper sensor corroborated with those obtained by qRT-PCR. The new paper-based sensing method can be extended to the screening of many viruses (e.g. the human immunodeficiency virus, the human polyomavirus, the human papilloma virus, the adeno associated viruses, the enteroviruses) through the cysteine residues of their capsid proteins. The new method has strong potential for screening viruses at pathology labs and in remote areas that lacks advanced scientific infrastructure. Further clinical studies are warranted to validate the new sensing method. [Display omitted] • A proof of concept on an optical sensor for screening viruses through the capsid proteins. • The sensor was used to detect SARS-CoV-2 through the S1 protein. • The free cysteine of the protein causes the sensor color to change spontaneously. • The sensor detected the protein in biological specimens down to 130 fg mL−1 • The sensing method is rapid when compared to qRT-PCR. [ABSTRACT FROM AUTHOR]
- Published
- 2022
- Full Text
- View/download PDF
48. Exosomes at the crossroad between therapeutic targets and therapy resistance in head and neck squamous cell carcinoma.
- Author
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Jangholi, Abolfazl, Müller Bark, Juliana, Kenny, Lizbeth, Vasani, Sarju, Rao, Sudha, Dolcetti, Riccardo, and Punyadeera, Chamindie
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- *
SQUAMOUS cell carcinoma , *EXOSOMES , *DRUG target , *EXTRACELLULAR vesicles , *DRUG carriers - Abstract
Head and neck squamous cell carcinomas (HNSCCs) are aggressive and clinically challenging tumours that require a multidisciplinary management approach. Despite significant therapy improvements, HNSCC patients have a poor prognosis with a 5-year survival rate of about 65%. As recently recognised key players in cancer, exosomes are extracellular vesicles (EVs) with a diameter of nearly 50–120 nm which transport information from one cell to another. Exosomes are actively involved in various aspects of tumour initiation, development, metastasis, immune regulation, therapy resistance, and therapeutic applications. However, current knowledge of the role of exosomes in the pathophysiological processes of HNSCC is still in its infancy, and additional studies are needed. In this review, we summarise and discuss the relevance of exosomes in mediating local immunosuppression and therapy resistance of HNSCC. We also review the most recent studies that have explored the therapeutic potential of exosomes as cancer vaccines, drug carriers or tools to reverse the drug resistance of HNSCC. • Head and neck squamous cell carcinomas (HNSCCs) are one of the most malignant tumours due to poor prognosis, high recurrence rates, and therapy resistance. • HNSCC-derived extracellular vesicles (EVs), especially exosomes, are abundant in the tumour microenvironment and play an active role in mediating signal transduction through delivering different biomolecules. • Exosomes have the potential to induce the resistance of HNSCC to both chemotherapy and radiotherapy as well as to modulate the antitumor activities of immune cells. • Due to their chemical structures and unique characteristics, exosomes are attractive candidates to devise novel treatment alternatives to prevent metastasis and overcome therapy resistance [ABSTRACT FROM AUTHOR]
- Published
- 2022
- Full Text
- View/download PDF
49. Current trends in the etiology and diagnosis of HPV-related head and neck cancers.
- Author
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Chai, Ryan C., Lambie, Duncan, Verma, Mukesh, and Punyadeera, Chamindie
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PAPILLOMAVIRUSES , *SQUAMOUS cell carcinoma , *ETIOLOGY of diseases , *HEAD & neck cancer , *ETIOLOGY of cancer , *NONINVASIVE diagnostic tests , *MORTALITY , *BIOMARKERS - Abstract
Human papilloma virus ( HPV) infection is a major risk factor for a distinct subset of head and neck squamous cell carcinoma ( HNSCC). The current review summarizes the epidemiology of HNSCC and the disease burden, the infectious cycle of HPV, the roles of viral oncoproteins, E6 and E7, and the downstream cellular events that lead to malignant transformation. Current techniques for the clinical diagnosis of HPV-associated HNSCC will also be discussed, that is, the detection of HPV DNA, RNA, and the HPV surrogate marker, p16 in tumor tissues, as well as HPV-specific antibodies in serum. Such methods do not allow for the early detection of HPV-associated HNSCC and most cases are at an advanced stage upon diagnosis. Novel noninvasive approaches using oral fluid, a clinically relevant biological fluid, allow for the detection of HPV and cellular alterations in infected cells, which may aid in the early detection and HPV-typing of HNSCC tumors. Noninvasive diagnostic methods will enable early detection and intervention, leading to a significant reduction in mortality and morbidity associated with HNSCC. [ABSTRACT FROM AUTHOR]
- Published
- 2015
- Full Text
- View/download PDF
50. A multimarker approach to diagnose and stratify heart failure.
- Author
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Yunxia Wan, Xi Xhang, Atherton, John J., Kostner, Karam, Dimeski, Goce, and Punyadeera, Chamindie
- Subjects
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HEART failure , *N-terminal residues , *C-terminal residues , *BRAIN natriuretic factor , *IMMUNOGLOBULINS , *BIOLOGICAL assay , *DIAGNOSIS - Abstract
Background We have previously demonstrated that circulating NT-proBNP is truncated at the N and C termini. Aims of this study are three-fold: firstly to determine whether the NT-proBNP levels correlate with NYHA functional classes when measuring with different antibody pairs; secondly to evaluate the diagnostic potential of ProBNP and; thirdly to investigate whether combining NT-proBNP assays with or without ProBNP would lead to better diagnostic accuracies. Methods Plasma samples were collected from healthy controls (n = 52) and HF patients (n = 46). Customized AlphaLISA(r) immunoassays were developed and validated to measure the concentrations of proBNP and NT-proBNP (with antibodies targeting 13-45, 13-76, 28-76). The diagnostic performance and predictive value of proBNP and NT-proBNP assays and their combinations were evaluated. Results Plasma proBNP assay showed acceptable diagnostic performance. NT-proBNP13-76 assay is useful in diagnosing and stratifying HF patients. The diagnostic performance of NT-proBNP13-76 demonstrated improvement over commercial NT-proBNP tests. The combination of NT-proBNP13-76 with NT-proBNP28-76 assays gave the best diagnostic assay performance. Conclusion Our results demonstrate that while there is major heterogeneity in circulating NT-proBNP, specific epitopes of the peptides are extraordinarily stable, providing ideal targets for clinically useful diagnostic assays. Future new clinical diagnostic clinical trials should include a multimarker approach rather than using a single marker to diagnose HF. [ABSTRACT FROM AUTHOR]
- Published
- 2015
- Full Text
- View/download PDF
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