Analytical, Diagnostic and Therapeutic Techniques and Equipment Commons^™

Improved Protocols For Vibrational Spectroscopic Analysis Of Body Fluids, Franck Bonnier, François Petitjean, Matthew Baker, Hugh Byrne

Articles

The applications of vibrational spectroscopy to the examination of human blood serum are explored. Although FTIR spectra can be recorded in aqueous solutions at (gelatin) concentrations as low as 100mg/L, the high-wavenumber region remains obscured by water absorption. Using Raman spectroscopy, high quality spectra of gelatine solutions as low as 10mg/L can be achieved, also covering the high-wavenumber regions. In human serum, spectral profiles are weak and partially obscured by water features. Dried deposits are shown to be physically and chemically inhomogeneous resulting in reduced measurement reproducibility. Concentration of the serum using commercially available centrifugal filter devices results in an …

Comparison Of Semi-Automated And Manual Measurements Of Carotid Intima-Media Thickening, Oscar Mac Ananey, G Mellotte, V Maher

Articles

Carotid intima-media thickening (CIMT) is a marker of both arteriosclerotic and atherosclerotic risks. Technological advances have semiautomated CIMT image acquisition and quantification. Studies comparing manual and automated methods have yielded conflicting results possibly due to plaque inclusion in measurements. Low atherosclerotic risk subjects (n = 126) were recruited to minimise the effect of focal atherosclerotic lesions on CIMT variability. CIMT was assessed by high-resolution B-mode ultrasound (Philips HDX7E, Phillips, UK) images of the common carotid artery using both manual and semiautomated methods (QLAB, Phillips, UK). Intraclass correlation coefficient (ICC) and the mean differences of paired measurements (Bland-Altman method) were used …

Raman Spectroscopic Analysis Of Human Skin Tissue Sections Ex-Vivo: Evaluation Of The Effects Of Tissue Processing And Dewaxing, Syed Mehmood Ali, Franck Bonnier, Ali Tfayli, Helen Lambkin, Kathleen Flynn, Vincent Mcdonagh, Claragh Healy, Thomas Lee, Fiona Lyng, Hugh Byrne

Articles

Raman spectroscopy coupled with K-means clustering analysis (KMCA) is employed to elucidate the biochemical structure of human skin tissue sections, and the effects of tissue processing. Both hand and thigh sections of human cadavers were analysed in their unprocessed and formalin fixed paraffin processed (FFPP) and subsequently dewaxed forms. In unprocessed sections, KMCA reveals clear differentiation of the stratum corneum, intermediate underlying epithelium and dermal layers for sections from both anatomical sites. The stratum corneum is seen to be relatively rich in lipidic content; the spectrum of the subjacent layers is strongly influenced by the presence of melanin, while that …

Detecting Point-Like Sources Of Unknown Frequency Spectra, Raffaele Solimene, Giuseppe Ruvio, Angela Dell’Aversano, Antonio Cuccaro, Max Ammann, Rocco Pierri

Articles

The problem of detecting point-like sources whose frequency spectrum is unknown is addressed. Limitations of single-frequency approaches are identified by analytical as well as numerical arguments. To overcome these limits, different multifrequency approaches which combine frequency data incoherently are compared. In particular, a novel multifrequency MUSIC-like algorithm based on interferometric concepts is proposed. Results show that the algorithm outperforms other methods under comparison.

Electric Field Standing Wave Effects In Ft-Ir Transflection Spectra Of Biological Tissue Sections: Simulated Models Of Experimental Variability, Tomasz P. Wrobel, Barbara Wajnchold, Hugh Byrne, Malgorzata Baranska

Articles

The so-called electric field standing wave effect (EFSW) has recently been demonstrated to significantly distort FT-IR spectra acquired in a transflection mode, both experimentally and in simulated models, bringing into question the appropriateness of the technique for sample characterization, particularly in the field of spectroscopy of biological materials. The predicted effects are most notable in the regime where the sample thickness is comparable to the source wavelength. In this work, the model is extended to sample thicknesses more representative of biological tissue sections and to include typical experimental factors which are demonstrated to reduce the predicted effects. These include integration …

A Comparison Of Raman, Ftir And Atr-Ftir Micro Spectroscopy For Imaging Human Skin Tissue Sections., Syed Mehmood Ali, Franck Bonnier, Helen Lambkin, Kathleen Flynn, V Mcdonagh, Claragh Healy, T C. Lee, Fiona Lyng, Hugh Byrne

Articles

Raman and Infrared absorption spectroscopies are compared for the analysis of human hand skin tissue sections. The tissue sections have been formalin fixed and paraffin processed, and subsequently dewaxed. Fourier Transform infrared (FTIR) spectra are preprocessed using the resonant Mie – extended multiplicative scattering algorithm to remove spectral artefacts. FTIR images of resolution 4cm^-1, analysed using K-means cluster analysis, reveal the double layer structure of the dermis and epidermis, but no further layer differentiation is achieved using the higher spatial resolution of the Attenuated Total Reflection imaging or improved spectral resolution of 2cm^-1. At comparable spectral …

Raman Spectroscopy In Nanomedicine: Current Status And Future Perspectives, Hugh Byrne, Mark Keating

Articles

Raman spectroscopy is a branch of vibration spectroscopy which is capable of probing the chemical composition of materials. Recent advances in Raman microscopy have added significantly to the range of applications which now extend from medical diagnostics to exploring interfaces between biological organisms and nanomaterials. In this review, Raman is introduced in a general context, highlighting some of the areas in which the technique has found success in the past, as well as some of the potential benefits it offers over other analytical modalities. The subset of Raman techniques which specifically probe the nanoscale, namely Surface Enhanced and Tip Enhanced …

Raman Micro Spectroscopy Study Of The Interaction Of Vincristine With A549 Cells Supported By Expression Analysis Of Bcl-2 Protein, Haq Nawaz, Amaya Garcia, Aidan Meade, Fiona Lyng, Hugh Byrne

Articles

Understanding the interaction of anticancer drugs with model cell lines is important to elucidate the mode of action of these drugs as well as to develop cost effective and rapid screening methods. Raman spectroscopy has been demonstrated to be a valuable technique for high throughput, noninvasive analysis. The interaction of vincristine with a human lung adenocarcinoma cell line (A549)was investigated using Raman micro spectroscopy. The results were correlated with parallel measurements from the MTT cytotoxicity assay, which yielded an IC₅₀ value of 0.10 ± 0.03 μM. The Raman spectral data acquired from vincristine treated A549 cells was analysed to …

Assessment Of An Osteoblast-Like Cell Line As A Model For Human Primary Osteoblasts Using Raman Spectroscopy, Lindsay Mcmanus, Franck Bonnier, George Burke, Brian Meenan, Adrian Boyd, Hugh Byrne

Articles

Raman spectroscopy is employed to determine the suitability of the U20S osteoblast-like cell line for use as a model for human primary osteoblasts, with emphasis on the ability of these cell types to replicate their tissue of origin. It was found that both cell types demonstrated early stage mineral deposition that followed significantly different growth patterns. Analysis of the growth pattern and spectral data from primary cells revealed increasing bone quality ratios and a high crystallinity, consistent with previous reports. Conversely the investigation of the U20S osteoblast-like cell line provided evidence of dense multilayered mineralised regions that corresponded more closely …

Analysis Of Human Skin Tissue By Raman Microspectroscopy: Dealing With The Background, Franck Bonnier, Syed Mehmood Ali, Peter Knief, Helen Lambkin, Kathleen Flynn, Vincent Mcdonagh, Claragh Healy, Thomas Lee, Fiona Lyng, Hugh Byrne

Articles

Raman microspectroscopy is widely used for molecular characterisation of tissue samples. Nevertheless, when working in vitro on tissue sections, the presence of a broad background to the spectra remains problematic and its removal requires advanced methods for pre-processing of the data. To date, research efforts have been primarily devoted to development of techniques of statistical analysis to extract the relevant information contained in the spectra. However, few attempts have been made to understand the origin of the background and to improve the protocols used for the collection of Raman spectra that could lead to the reduction or elimination of the …

Comparison Of Subcellular Responses For The Evaluation And Prediction Of The Chemotherapeutic Response To Cisplatin In Lung Adenocarcinoma Using Raman Spectroscopy, Haq Nawaz, Franck Bonnier, Aidan Meade, Fiona Lyng, Hugh Byrne

Articles

Confocal Raman Micro spectroscopy (CRM) is employed to examine the chemical and physiological effects of anticancer agents, using cisplatin and A549 adenocarcinoma cells as a model compound and test system respectively. Spectral responses of the membrane and cytoplasm of the cell are analysed independently and the results are compared to previously reported spectroscopic studies of the nucleus. Moreover, Raman spectra from the proteins extracted from the control and exposed samples are acquired and analysed to confirm the origin of the molecular changes of the cell membrane and cytoplasm of the A549 cells. Multivariate data analysis techniques including Principal Component Analysis …

Development And Validation Of A Rapid Multi-Class Method For The Confirmation Of Fourteen Prohibited Medicinal Additives In Pig And Poultry Compound Feed By Liquid Chromatography Tandem Mass Spectrometry, Mark Cronly, Patrice Behan, Barry Foley, Liam Regan, Marella Gallagher, Sean Earley, Paula Shearan, Edward Malone

Articles

A confirmatory method has been developed to allow for the analysis of fourteen prohibited medicinal additives in pig and poultry compound feed. These compounds are prohibited for use as feed additives although some are still authorised for use in medicated feed. Feed samples are extracted by acetonitrile with addition of sodium sulphate. The extracts undergo a hexane wash to aid with sample purification. The extracts are then evaporated to dryness and reconstituted in initial mobile phase. The samples undergo an ultracentrifugation step prior to injection onto the LC-MS/MS system and are analysed in a run time of 26 minutes. The …

A Monocular Marker-Free Gait Measurement System, Jane Courtney, Annraoi M. De Paor

Articles

This paper presents a new, user-friendly, portable motion capture and gait analysis system for capturing and analyzing human gait, designed as a telemedicine tool to monitor remotely the progress of patients through treatment. The system requires minimal user input and simple single-camera filming (which can be acquired from a basic webcam) making it very accessible to nontechnical, nonclinical personnel. This system can allow gait studies to acquire a much larger data set and allow trained gait analysts to focus their skills on the interpretation phase of gait analysis. The design uses a novel motion capture method derived from spatiotemporal segmentation …

Rapid Multi-Class Multi-Residue Method For The Confirmation Of Chloramphenicol And Eleven Nitroimidazoles In Milk And Honey By Liquid Chromatography Tandem Mass Spectrometry, Mark Cronly, Patrice Behan, Barry Foley, Liam Regan, Sheila Martin, Michael Doyle, Edward Malone

Articles

A confirmatory method has been developed to allow for the analysis of eleven nitroimidazoles and also chloramphenicol in milk and honey samples. These compounds are classified as A6 compounds in annex IV of council regulation 2377/90 and therefore prohibited for the use in animal husbandry. Milk samples are extracted by acetonitrile with addition of NaCl, while honey samples are first dissolved in water before a similar extraction. Honey extracts undergo a hexane wash to remove impurities. Both milk and honey extracts are evaporated to dryness and reconstituted in initial mobile phase. These are then injected onto an LC-MS/MS system and …

An Optical Machine Vision System For Applications In Cytopathology, Jonathan Blackledge, Dmitry Dubovitskiy

Articles

This paper discusses a new approach to the processes of object detection, recognition and classification in a digital image focusing on problem in Cytopathology. A unique self learning procedure is presented in order to incorporate expert knowledge. The classification method is based on the application of a set of features which includes fractal parameters such as the Lacunarity and Fourier dimension. Thus, the approach includes the characterisation of an object in terms of its fractal properties and texture characteristics. The principal issues associated with object recognition are presented which include the basic model and segmentation algorithms. The self-learning procedure for …

Fractional Anisotropic Diffusion For Noise Reduction In Magnetic Resonance Images, Jonathan Blackledge, Matthew Blackledge

Articles

We extend the method of anisotropic diffusion for noise reduction in digital images to the case when the diffusion processes are non-Gaussian and Levy distributed. This yields a fractional diffusion equation characterised by the Levy index. A solution to this equation is considered and a numerical algorithm developed. The algorithm is then applied as a case study to the problem of reducing noise in magnetic resonance imaging. The focus of this study is on diffusion weighted images which have low signal-to-noise ratios.

Platelet Alpha- And Beta- Secretase Activities Are Not Significantly Affected By Dementia Or Mild Cognitive Impairment In Swedish Patients., Paul Gorham, Natalie Bark, Steve Meaney, Ingemar Bjorkhem, Milita Crisby

Articles

The processing of the Amyloid Precursor Protein (APP) is a critical event in the formation of amyloid plaques which are composed of the 4kDa amyloid beta-peptide (Abeta). Processing of APP occurs through a non-amyloidogenic pathway, mediated by initial alpha-secretase cleavage or through an amyloidogenic pathway via sequential cleavage by beta- and gamma-secretase enzymes, which produces Abeta peptides. Currently, the diagnosis of probable or possible Alzheimer's disease (AD) is primarily based on neuropsychological and neuroradiological assessment. Recent reports indicate that platelet beta-secretase activity is moderately increased in patients with AD and mild cognitive impairment (MCI). To our knowledge platelet alpha-secretase activity …

Hmgb1 Mediates Endogenous Tlr2 Activation And Brain Tumor Regression, James Curtin, Naiyou Liu, Marianela Candolfi, Weidong Xiong, Hikmat Assi, Kader Yagiz, Matthew Edwards, Kathrin Michelsen, Kurt Kroeger, Chunyan Liu, Akm Ghulam Muhammad, Mary Clark, Moshe Arditi, Begonya Comin-Anduix, Antoni Ribas, Pedro Lowenstein, Maria Castro

Articles

BACKGROUND: Glioblastoma multiforme (GBM) is the most aggressive primary brain tumor that carries a 5-y survival rate of 5%. Attempts at eliciting a clinically relevant anti-GBM immune response in brain tumor patients have met with limited success, which is due to brain immune privilege, tumor immune evasion, and a paucity of dendritic cells (DCs) within the central nervous system. Herein we uncovered a novel pathway for the activation of an effective anti-GBM immune response mediated by high-mobility-group box 1 (HMGB1), an alarmin protein released from dying tumor cells, which acts as an endogenous ligand for Toll-like receptor 2 (TLR2) signaling …

Treg Depletion Inhibits Efficacy Of Cancer Immunotherapy: Implications For Clinical Trials., James Curtin, Marianela Candolfi, Tamer Fakhouri, Chunyan Liu, Anderson Alden, Matthew Edwards, Pedro Lowenstein, Maria Castro

Articles

BACKGROUND: Regulatory T lymphocytes (Treg) infiltrate human glioblastoma (GBM); are involved in tumor progression and correlate with tumor grade. Transient elimination of Tregs using CD25 depleting antibodies (PC61) has been found to mediate GBM regression in preclinical models of brain tumors. Clinical trials that combine Treg depletion with tumor vaccination are underway to determine whether transient Treg depletion can enhance anti-tumor immune responses and improve long term survival in cancer patients. FINDINGS: Using a syngeneic intracrabial glioblastoma (GBM) mouse model we show that systemic depletion of Tregs 15 days after tumor implantation using PC61 resulted in a decrease in Tregs …

A Surface Inspection Machine Vision System That Includes Fractal Texture Analysis, Jonathan Blackledge, Dmitry Dubovitskiy

Articles

The detection, recognition and classification of features in a digital image is an important component of quality control systems in production and process engineering and industrial systems monitoring, in general. In this paper, a new pattern recognition system is presented that has been designed for the specific task of monitoring the quality of sheet-steel production in a rolling mill. The system is based on using both the Euclidean and Fractal geometric properties of an imaged object to develop training data that is used in conjunction with a supervised learning procedure based on the application of a fuzzy inference engine. Thus, …

Object Detection And Classification With Applications To Skin Cancer Screening, Jonathan Blackledge, Dmitryi Dubovitskiy

Articles

This paper discusses a new approach to the processes of object detection, recognition and classification in a digital image. The classification method is based on the application of a set of features which include fractal parameters such as the Lacunarity and Fractal Dimension. Thus, the approach used, incorporates the characterisation of an object in terms of its texture.

The principal issues associated with object recognition are presented which includes two novel fast segmentation algorithms for which C++ code is provided. The self-learning procedure for designing a decision making engine using fuzzy logic and membership function theory is also presented and …

Adenoviral Mediated Gene Transfer Into The Dog Brain In Vivo, Marianela Candolfi, Kurt Kroeger, Elizabeth Pluhar, Chunyan Liu, Carlos Barcia, Josee Bergeron, Mariana Puntel, James Curtin, Elizabeth Mcniel, Andrew Freese, John Ohlfest, Peter Moore, William Kuoy, Pedro Lowenstein, Maria Castro

Articles

OBJECTIVE: Glioblastoma multiforme (GBM) is a devastating brain tumor for which there is no cure. Adenoviral-mediated transfer of conditional cytotoxic (herpes simplex virus [HSV] 1-derived thymidine kinase [TK]) and immunostimulatory (Fms-like tyrosine kinase 3 ligand [Flt3L]) transgenes elicited immune-mediated long-term survival in a syngeneic intracranial GBM model in rodents. However, the lack of a large GBM animal model makes it difficult to predict the outcome of therapies in humans. Dogs develop spontaneous GBM that closely resemble the human disease; therefore, they constitute an excellent large animal model. We assayed the transduction efficiency of adenoviral vectors (Ads) encoding beta-galactosidase (betaGal), TK, …

Preliminary Results For A Monocular Marker-Free Gait Measurement System, Jane Courtney, Annraoi Depaor

Articles

This paper presents results from a novel monocular marker-free gait measurement system. The system was designed for physical and occupational therapists to monitor the progress of patients through therapy. It is based on a novel human motion capture method derived from model-based tracking. Testing is performed on two monocular, sagittal-view, sample gait videos – one with both the environment and the subject’s appearance and movement restricted and one in a natural environment with unrestricted clothing and motion. Results of the modelling, tracking and analysis stages are presented along with standard gait graphs and parameters.