Window into the mind:Advanced handheld spectroscopic eye-safe technology for point-of-care neurodiagnostic

Traumatic brain injury (TBI), a major cause of morbidity and mortality worldwide, is hard to diagnose at the point of care with patients often exhibiting no clinical symptoms. There is an urgent need for rapid point-of-care diagnostics to enable timely intervention. We have developed a technology for rapid acquisition of molecular fingerprints of TBI biochemistry to safely measure proxies for cerebral injury through the eye, providing a path toward noninvasive point-of-care neurodiagnostics using simultaneous Raman spectroscopy and fundus imaging of the neuroretina. Detection of endogenous neuromarkers in porcine eyes' posterior revealed enhancement of high-wave number bands, clearly distinguishing TBI and healthy cohorts, classified via artificial neural network algorithm for automated data interpretation. Clinically, translating into reduced specialist support, this markedly improves the speed of diagnosis. Designed as a hand-held cost-effective technology, it can allow clinicians to rapidly assess TBI at the point of care and identify long-term changes in brain biochemistry in acute or chronic neurodiseases.</p

Advanced Tuneable Micronanoplatforms for Sensitive and Selective Multiplexed Spectroscopic Sensing via Electro-Hydrodynamic Surface Molecular Lithography

Micro- and nanopatterning of materials, one of the cornerstones of emerging technologies, has transformed research capabilities in lab-on-a-chip diagnostics. Herein, a micro- and nanolithographic method is developed, enabling structuring materials at the submicron scale, which can, in turn, accelerate the development of miniaturized platform technologies and biomedical sensors. Underpinning it is the advanced electro-hydrodynamic surface molecular lithography, via inducing interfacial instabilities produces micro- and nanostructured substrates, uniquely integrated with synthetic surface recognition. This approach enables the manufacture of design patterns with tuneable feature sizes, which are functionalized via synthetic nanochemistry for highly sensitive, selective, rapid molecular sensing. The development of a high-precision piezoelectric lithographic rig enables reproducible substrate fabrication with optimum signal enhancement optimized for functionalization with capture molecules on each micro- and nanostructured array. This facilitates spatial separation, which during the spectroscopic sensing, enables multiplexed measurement of target molecules, establishing the detection at minute concentrations. Subsequently, this nano-plasmonic lab-on-a-chip combined with the unconventional computational classification algorithm and surface enhanced Raman spectroscopy, aimed to address the challenges associated with timely point-of-care detection of disease-indicative biomarkers, is utilized in validation assay for multiplex detection of traumatic brain injury indicative glycan biomarkers, demonstrating straightforward and cost-effective micro- and nanoplatforms for accurate detection.</p

A Review on Advances in Intra-operative Imaging for Surgery and Therapy: Imagining the Operating Room of the Future

none4openZaffino, Paolo; Moccia, Sara; De Momi, Elena; Spadea, Maria FrancescaZaffino, Paolo; Moccia, Sara; De Momi, Elena; Spadea, Maria Francesc

From Raman to SESORRS : moving deeper into cancer detection and treatment monitoring

Raman spectroscopy is a non-invasive technique that allows specific chemical information to be obtained from various types of sample. The detailed molecular information that is present in Raman spectra permits monitoring of biochemical changes that occur in diseases, such as cancer, and can be used for the early detection and diagnosis of the disease, for monitoring treatment, and to distinguish between cancerous and non-cancerous biological samples. Several techniques have been developed to enhance the capabilities of Raman spectroscopy by improving detection sensitivity, reducing imaging times and increasing the potential applicability for in vivo analysis. The different Raman techniques each have their own advantages that can accommodate the alternative detection formats, allowing the techniques to be applied in several ways for the detection and diagnosis of cancer. This feature article discusses the various forms of Raman spectroscopy, how they have been applied for cancer detection, and the adaptation of the techniques towards their use for in vivo cancer detection and in clinical diagnostics. Despite the advances in Raman spectroscopy, the clinical application of the technique is still limited and certain challenges must be overcome to enable clinical translation. We provide an outlook on the future of the techniques in this area and what we believe is required to allow the potential of Raman spectroscopy to be achieved for clinical cancer diagnostics

Geographical races of the honeybees (Apis Mellifera L.) of the Northern regions of Ethiopia

The ideal climatic conditions and diversity of floral resources allow Ethiopia to sustain millions of honeybee colonies. Beekeeping is widely practiced and plays a significant role in the livelihood of the farming community of the country. Despite this, information on the types of geographical races of honeybees, their behavioural characters and the related ecological factors are not established. In this work an attempt was made to characterise the honeybee populations of the northern parts of Ethiopia based on the analyses of morphological, behavioural and ecological characteristics. For morphological analysis, 155 honeybee colony samples from 31 localities were collected. 20 worker honeybees per colony sample totalled 3100 individual worker honeybees and thirteen morphological characters per bee, a total of 40 300 measurements were recorded following Ruttner's (1988) protocols. The behavioural characters such as reproductive swarming, migration, temperament and colony seasonal cycles were assessed based on survey work of a pre-structured questionnaire. Various univariate and multivariate statistical methods were employed to analyse the data. The principal components and step-wise linear discriminant analyses revealed the existence of four discrete morphoclusters or geographical races: A. m. jemenitica from eastern, northwestern and northern arid and semi-arid lowlands, A. m. scutellata from the western humid midlands, A. m. bandasii from sub-moist central highlands and A. m. monticola from the northern mountainous parts of the study areas. These different geographical races were found to be distinctively distributed in the different ecological parts of the study area. High intercolonial and intracolonial variances were observed in all localities, however particularly high values were obtained in areas of transitions between ecological zones indicating a region of gene flow or zones of hybridisation among the statistically defined populations. Such high variances were observed mostly in areas where A. m. jemenitica borders the other geographical races. Moreover. distinct behavioural variations were also noted among these geographical races. Generally. the highland and mountainous honeybees A. m. bandasii and A. m. monticola are relatively larger in body size. have less tendency to swarm. less inclination to migrate and are relatively gentle while the converse traits were noted for the lowlands and midlands honeybees A. m. jemenitica and A. m. scutellata. Both morphological and behavioural variations were highly correlated with environmental factors and the variations seem to be the result of long adaptation to the interrelated ecological factors in their respective areas. Variations in reproductive swarming periods were also noted among these honeybee populations as a result of variations in rainfall pattern. altitude and temperature in their respective ecological areas which are believed to alter the honeybee colonies' seasonal cycles and leads to partial temporal reproductive isolation among these different honeybee populations

MOLECULAR ANALYSIS OF CANCER PROGRESSION WITH LABEL-FREE RAMAN SPECTROSCOPY

Due to its ability to probe water-containing samples using visible and near-infrared frequencies with high chemical specificity, Raman spectroscopy is an attractive tool for label-free investigation of biological samples. While Raman spectroscopy has been leveraged for exploratory studies in clinical cancer diagnostics, only limited studies have used it to understand the molecular mechanisms driving key characteristics of cancer progression. In this thesis, we present three progressively complex applications of Raman spectroscopy that take advantage of its specificity and synergistic combination with plasmonic nanoparticles and multivariate data analysis for molecular study of cancer. First, we used Au@SiO2 shell-isolated nanoparticle-enhanced Raman spectroscopy (SHINERS) to investigate the roles of microcalcification status and the composition of tumor microenvironment in breast tissue for identification of a range of breast pathologies. We developed a partial least squares-discriminant analysis-based classifier to correlate the spectra with their pathology to obtain high prediction accuracy. A parallel investigation of the genetic drivers of microcalcification formation in breast cancer cells revealed that stable silencing of the Osteopontin gene decreased the formation of hydroxyapatite in breast cancer cells and reduced their migration. Next, we demonstrated the ability to detect premetastatic changes in the lungs of mice bearing breast tumors, in advance of tumor cell seeding, using Raman spectroscopy and multivariate data analysis. Our measurements showed reliable differences in the collagen and proteoglycan features of the premetastatic lungs which uniquely identify the metastatic potential of the primary tumor. Consistent with histological assessment, our results hint at a continuous premetastatic niche formation model dependent on the metastatic potential of primary tumor. Finally, we exploited Raman mapping to elucidate radiation therapy-induced biomolecular changes in murine tumors and uncovered latent microenvironmental differences between treatment-resistant and -sensitive tumors. We used multivariate curve resolution-alternating least squares (MCR-ALS) and support vector machine (SVM) to quantify biomolecular differences in the tumor microenvironment and constructed classification models to predict therapy outcome and resistance. We found significant differences in lipid and collagen content between unirradiated and irradiated tumors. Taken together, these studies pave the way for applications of Raman spectroscopy beyond clinical diagnostics such as metastatic risk assessment and treatment monitoring

Study of genetic structure of honeybee populations Apis mellifera

Honeybee populations from six different areas of Greece and Cyprus were studied which may correspond to different races according to Ruttner morphometry analysis (1988). The aim is to study the genetic structure of these populations to examine their phylogenetic relationships and to estimate the gene flow because of migratory beekeeping and commercial breeding. The methods that have been used were starch gel electrophoresis on different enzymatic systems (a GPDH AO MDH ADH LAP SOD ALP ACPH ME and EST) which correspond at 12 hypothetical genetic loci and restriction fragment length polymorphisms of three mtDNA gene (16srDNA COI and ND5) segments. From the enzymic data using the BIOSYS 1 and PHYLIP software packages Nei’s genetic distance was calculated and phylogenetic trees were constructed using the UPGMA Neighbor Joining and Wagner parsimony methods all trees support the assumption that the more distant population is that of Cyprus From mtDNA analysis the nucleotide divergence among the studied populations was calculated using REAP and PHYLIP programs and phylogenetic trees constructed by the UPGMA Neighbor Joining and Dollo parsimony methods. Also a computing method has been developed to define the most probable order of DNA gene fragments and to identify the sequence of the DNA end fragments. All trees obtained revealed that the more distant population is the Macedonian one. Our data are not in coincidence with Ruttner s morphometry analysis they reflect a systematic attempt to the genetic study of Greek bees populations and could be useful as regards the migratory beekeeping and commercial breeding.Μελετήθηκε η γενετική δομή και οι φυλογενετικές σχέσεις πληθυσμών μελισσών από έξι διαφορετικές περιοχές της Ελλάδας και της Κύπρου (A m adami A m carnica A m macedonica A m cecropia A m cypria κατά Ruttner 1988). Σκοπός είναι η διαπίστωση τυχόν γενετικών διαφοροποιήσεων των πληθυσμών και να εκτιμηθεί η έκταση της μείξης των φυλών λόγω μετακινήσεων και αγοραπωλησιών. Χρησιμοποιήθηκε η τεχνική της ηλεκτροφόρησης αμύλου των ενζυμικών συστημάτων a GPDH ΑΟ MDH ADH LAP SOD ALP ACPH ME και EST και η μελέτη πολυμορφισμών τριών γονιδιακών τμημάτων του mtDNA (16srDNA COI και ND5) μετά από πέψεις με ένζυμα περιορισμού. Τα ενζυμικά πρότυπα ερμηνεύθηκαν με βάση την ύπαρξη 12 γενετικών τόπων υπολογίστηκαν οι γενετικές αποστάσεις και κατασκευάστηκαν φυλογενετικά δένδρα με τις μεθόδους UPGMA Neighbor Joining και φειδωλότητας του Wagner. Κοινό συμπέρασμα είναι ότι ο πληθυσμός της Κύπρου είναι γενετικά ο πιο απομακρυσμένος. Από τα δεδομένα του mtDNA υπολογίστηκαν οι συντελεστές νουκλεοτιδικών υποκαταστάσεων και κατασκευάστηκαν φυλογενετικά δένδρα με τις μεθόδους Neighbor Joining UPGMA και φειδωλότητας του Dollo. Σε όλες τις περιπτώσεις και σε συνδυασμό με την ύπαρξη διαγνωστικών ένζυμων ο πληθυσμός της Μακεδονίας εμφανίζεται ως ο πιο απομακρυσμένος πληθυσμός. Τα αποτελέσματα της έρευνας δείχνουν ότι έχουν αυξηθεί οι μετακινήσεις και αγοροπωλησίες μελισσιών η/και βασιλισσών με συνέπεια μεταβολές στη γενετική δομή της ελληνικής μέλισσας