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Raman spectroscopy (RS) is widely used as a non-invasive technique in screening for the diagnosis of oral cancer. The potential of this optical technique for several biomedical applications has been proved. This work studies the efficacy of RS in detecting oral cancer using sub-site-wise differentiation. A total of 80 samples (44 tumor and 36 normal) were cryopreserved from three different sub-sites: The tongue, the buccal mucosa, and the gingiva of the oral mucosa during surgery. Linear discriminant analysis (LDA) and quadratic discriminant analysis (QDA) were used with principal component analysis (PCA) to classify the samples and the classifications were validated by leave-one-out-cross-validation (LOOCV) and k-fold cross-validation methods. The normal and tumor tissues were differentiated under the PCA-LDA model with an accuracy of 81.25% (sensitivity: 77.27%, specificity: 86.11%). The PCA-QDA classifier model differentiated these tissues with an accuracy of 87.5% (sensitivity: 90.90%, specificity: 83.33%). The PCA-QDA classifier model outperformed the PCA-LDA-based classifier. The model studies revealed that protein, amino acid, and beta-carotene variations are the main biomolecular difference markers for detecting oral cancer.
We believe this unmet need can be addressed by combining advancements in vibrational spectroscopy (Raman Spectroscopy, RS; Near Infra-Red Spectroscopy, NIRS; and mid infrared spectroscopy, MIRS) with computational methods (machine learning). These technologies have potential to provide non-perturbative, rapid and label-free tissue assessment of morphology and metabolism [4,5,6,7] at the molecular level [8, 9]. In the cardiovascular space, studies from animal or preserved tissue [4, 7, 10,11,12,13,14,15,16] demonstrate that these techniques can quantify myocardial fibrosis by exploring collagen subtypes [17,18,19,20], cross linking [21, 22] and distribution [20, 23]. The techniques are complementary, with RS techniques being more sensitive to metabolic data at a molecular level [24], and infrared-based techniques (such as MIRS, NIRS) better equipped to assess morphological changes [24]. Advances in machine learning have significantly reduced the processing time for analysing these data, and bear potential for real-time diagnoses. Clinical translation is hindered by current studies being restricted to one modality of spectroscopy (RS, Mid IR or NIRS) and have not yet used point-of-care instruments on human tissue.
In this report, we combine RS and NIRS scans of cardiac tissue to obtain non-invasive multimodal spectroscopic signatures (MSS) and use machine learning (ML) to compare its accuracy to conventional histopathology. These scans, which can be performed in 3 s, are iterative steps to generating a point-of-care and non-invasive morphological and metabolic diagnoses in heart disease.
This study was approved by the Human Resources and Ethics committee (HREC) at Austin Hospital, Heidelberg, Melbourne, Victoria (HREC/73660/Austin-2021). Approval of the acquisition of human tissue from organ donors was as part of the Australian Donation and Transplantation Biobank (HREC/4814/Austin-2019) and Donate Life Victoria (DLV) through the Australian Red Cross Lifeblood Health Human Research and Ethics Committee (Ethics 2019#08). Human tissue from explanted hearts was with USYD HREC 2021/122. Individual consent was obtained from either the patient or Senior Next Available next of Kin (SANOK) prior to accessing samples. Ethics approvals are in keeping with the Declaration of Helsinki.
Pathological human heart samples were obtained from the Sydney Heart Bank at the University of Sydney, New South Wales, Australia [25] from patient samples of dilated cardiomyopathy and ischaemic heart disease pathologies. Physiological control patient samples were prospectively collected from consecutive donors providing organs for transplantation in Victoria, Australia from the Australian Donation and Transplantation Biobank (ADTB) [26]. These were taken from hearts not utilised in transplantation, with the protocol having been described previously [26, 27]. For analysis, all fresh samples were thawed under standard laboratory conditions. All samples were de-identified and anonymously catalogued at the time of excision and source blinded for all subsequent analysis.
Algorithm for Multimodal Spectroscopic Signature acquisition from cardiac patient samples. Fresh tissue was acquired from 15 patients at time of cardiac transplantation, either from explanted dilated and ischaemic cardiomyopathy patients or healthy hearts at time of organ retrieval. Samples were scanned using separate Raman (3 scans per sample) and NIRS (4 scans per sample) instruments to acquire an MSS (combines the 7 scans per sample, 35 in total for each pathology), and then entered into machine learning (ML) algorithms to assess their diagnostic accuracy. H&E haematoxylin and eosin, PSR picrosirius red, VG Van Gieson. Created with BioRender.com
Study data were collected and managed using Research Electronic Data Capture (REDCap) electronic data capture tools hosted at ADTB. Analysis of clinical data was done using Stata v15.0 (StataCorp. 2017. Stata Statistical Software: Release 15. College Station, TX: StataCorp LLC), with clinical variables reported as either counts with corresponding percentages, or median averages with interquartile range (IQR).
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Dr. Shiv Sharma is a Tenured Research Professor at the Hawaii Institute of Geophysics and Planetology, School of Ocean and Earth Science and Technology (SOEST) at the University of Hawaii at Manoa. He is one of the Co-Principal Investigators for SuperCam on the Perseverance rover.
Serving on the Graduate Faculty of both the Department of Earth Sciences, SOEST; and in the Department of Electrical Engineering (EE), College of Engineering. Since 1998, serving on the Faculty of Global Environmental Science, an Undergraduate-Degree Program administered by the Dept. of Oceanography/SOEST, and also serving as a collaborator in the NASA Astrobiology Institute at UHM since 2010.
In the late 1990s/early 2000s, Shiv demonstrated that a pulsed laser coupled with time-gating could be used to take Raman measurements in the daylight, which was revolutionary in more than one way. In addition to being resistance to ambient light and thermal emissions, fluorescence, which is intense and often interferes with Raman signals, could also be avoided. Time-gated Raman spectroscopy synchronizes the pulsed laser source with the sensitive detector so that it can detect the fast Raman signals during the short laser pulses and avoid fluorescence emissions which have longer delays. Professor Sharma and Professor Angel published the first paper on the remote pulsed Raman system in 2002, showing that mineral samples on planetary surfaces could be analyzed up to 66 meters away.4
The predecessor of the Perseverance rover, Curiosity, only has the capability of performing laser induced breakdown spectroscopy (LIBS), which gives information about the atomic composition. The team at NASA wanted to learn more about the molecular structure of the samples, information which Raman spectroscopy could afford. In 2001, Dr. Sharma met Roger Wiens, the Principal Investigator of SuperCam at the Los Alamos National Laboratory. Dr. Sharma offered some advice regarding Raman spectra measurements of minerals with a pulsed laser. The experiments took off and so did their friendship.
Professor Stanley Michael Angel is a Carolina Trustee Professor and Fred M. Weissman Palmetto Chair in Chemical Ecology, Department of Chemistry and Biochemistry at the University of South Carolina. He currently works on the SuperCam team as a Scientific Research Collaborator and Scientific Payload Download Leader (sPDL).
Roger Wiens is the Principal Investigator of SuperCam and one of the co-investigators of SHERLOC. In 2016, the government of France knighted Wiens for his contribution in forging strong bonds between the French and American scientific communities.
Sanford Asher, a Distinguished Professor in the Department of Chemistry at the University of Pittsburgh, is involved with the development of the Raman spectrometer and the ultraviolet laser in SHERLOC.
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The exotic correlated phases that emerge in the low-dispersing moir bands of twisted van der Waals structures have opened new exciting opportunities in condensed-matter physics. These systems exhibit a rich phase diagram of novel physical phenomena and exotic correlated phases that emerge in the low-dispersing bands. Despite its popularity, the resulting mini-bands in the conduction band of MoS2 moir superlattices remained elusive so far. In this work, we resolve these spin-valley mini-bands via transport spectroscopy in micron-scale devices. The theoretical energy scales exhibit an astounding agreement with our experimental observations, which together with the behavior under thermal activation, suggest an electronic phase transition. These intriguing observations highlight the potential of twisted MoS2 structures as a playground to explore correlated electron states and associated phenomena.
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