Physics Commons^™

Determination Of Nanoparticle Localisation Within Subcellular Organelles In Vitro Using Raman Spectroscopy, Esen Efeoglu, Mark Keating, Jennifer Mcintyre, Alan Casey, Hugh Byrne

Articles

Ease of sample preparation, narrow spectral bandwidth and minimal influence from water are features of Raman spectroscopy which make it a powerful, label-free way to study a wide range of biological structures and phenomena. In this context, given the concerns over their toxicology arising from their increased production and use, evaluation of nanoparticle uptake and localisation in biological systems and determination of the mechanisms of subcellular interaction and trafficking can provide long-term solutions for nanotoxicology, and potential strategies for nanomedicine. In this study, Raman spectroscopy is explored to monitor the sequential trafficking of nanoparticles through subcellular organelles in-vitro and to …

Investigating The Role Of Shape On The Biological Impact Of Gold Nanoparticles In Vitro, Furong Tian, Hugh Byrne, Joao Conde, Tobias Stoeger, Martin Clift,, Alan Casey, Pablo Del Pino, Beatriz Pelaz, Barbara Rothen-Rutishauser,, Giovani Estrada, Jesús De La Fuente

Articles

Aim: To investigate the influence of gold nanoparticle (GNP) geometry on the biochemical response of Calu-3 epithelial cells.

Materials and Methods: Spherical, triangular and hexagonal GNPs were used. The GNP-cell interaction was assessed via atomic absorption spectroscopy (AAS) and transmission electron microscopy (TEM). The biochemical impact of GNPs was determined over 72hrs at [0.0001-1mg/mL].

Results: At 1mg/mL, hexagonal GNPs reduced Calu-3 viability below 60%, showed increased reactive oxygen species production and higher expression of pro-apoptotic markers. A cell mass burden of 1:2:12 as well as number of GNPs per cell (2:1:3) was observed for spherical:triangular:hexagonal GNPs.

Conclusion:

These findings do …

Vibrational Spectroscopic Studies To Elucidate The Structure Of Water At Biological Interfaces, Bahar Bahrani, Luke O'Neill, Hugh Byrne

Articles

In biological systems, water takes up to 80% of the volume inside a cell. This water solubilizes the biological macromolecules such as the DNA, proteins and lipids. Recent advancements have shown that the water at the interface of a lipid membrane is structured, as five layers of structured water have been found at this solvent cage. Steady state Raman spectroscopy of water in lipids was performed in an attempt to elucidate the structure of water at the biological interface. Deuterium oxide (heavy water) was employed to hydrate lipid biomolecules. The heavier deuterium atom shifts the molecular vibrations and renders them …

Spectropathology For The Next Generation: Quo Vadis?, Hugh Byrne, Malgorzata Baranska, Gerwin J. Pupples, Nick Stone, Bayden Wood, Kathleen M. Gough, Peter Lasch, Phil Heraud, Josep Sulé-Suso, Ganesh Sockalingum

Articles

Although the potential of vibrational spectroscopy for biomedical applications has been well demonstrated, translation into clinical practice has been relatively slow. This Editorial assesses the challenges facing the field and the potential way forward. While many technological challenges have been addressed to date, considerable effort is still required to gain acceptance of the techniques among the medical community, standardise protocols, extend to a clinically relevant scale, and ultimately assess the health economics underlying clinical deployment. National and international research networks can contribute much to technology development and standardisation. Ultimately, large-scale funding is required to engage in clinical trials and instrument …

Vibrational Microspectroscopy For Cancer Screening, Fiona Lyng, Ines Ramos, Ola Ibrahim, Hugh Byrne

Articles

Vibrational spectroscopy analyses vibrations within a molecule and can be used to characterise a molecular structure. Raman spectroscopy is one of the vibrational spectroscopic techniques, in which incident radiation is used to induce vibrations in the molecules of a sample, and the scattered radiation may be used to characterise the sample in a rapid and non-destructive manner. Infrared (IR) spectroscopy is a complementary vibrational spectroscopic technique based on the absorption of IR radiation by the sample. Molecules absorb specific frequencies of the incident light which are characteristic of their structure. IR and Raman spectroscopy are sensitive to subtle biochemical changes …

Multivariate Statistical Methodologies Applied In Biomedical Raman Spectroscopy: Assessing The Validity Of Partial Least Squares Regression Using Simulated Model Datasets, Mark E. Keating, Haq Nawaz, Franck Bonnier, Hugh Byrne

Articles

Raman spectroscopy is fast becoming a valuable analytical tool in a number of biomedical scenarios, most notably disease diagnostics. Importantly, the technique has also shown increasing promise in the assessment of drug interactions on a cellular and subcellular level, particularly when coupled with multivariate statistical analysis. However, an important consideration, both with Raman spectroscopy and the associated statistical methodologies, is the accuracy of these techniques and more specifically the sensitivities which can be achieved and ultimately the limits of detection of the various methods. The purpose of this study is thus the construction of a model simulated data set with …

Antimicrobial Properties Of Nano-Silver: A Cautionary Approach To Ionic Interference, Kate Sheehy, Alan Casey, Anna Murphy, Gordon Chambers

Articles

Hypothesis: Metallic nanoparticles such as nano-silver have found many applications as alternative antimicrobials in recent years. However methods for determining their proposed antimicrobial activity have received little attention to date. The disk diffusion assay is commonly used as a demonstration of antimicrobial properties and is a regular feature in synthetic nanoparticle papers. The aim of this study was to assess its effectiveness in demonstrating the ‘‘nanoparticle specific’’ antimicrobial properties in the absence of ionic contributions from unreacted reducing agents and or impurities. Experiments: The disk diffusion assay was carried out on a range of silver nanoparticles, both in-house synthesised and …

Competitive Evaluation Of Data Mining Algorithms For Use In Cassification Of Leukocyte Subtypes With Raman Microspectroscopy, Adrian Maguire, I. Vega-Carrascal, Jane Bryant, Lisa White, Orla Howe, Fiona Lyng, Aidan Meade

Articles

Raman microspectroscopy has been investigated for some time for use in label-free cell sorting devices. These approaches require coupling of the Raman spectrometer to complex data mining algorithms for identification of cellular subtypes such as the leukocyte subpopulations of lymphocytes and monocytes. In this study, three distinct multivariate classification approaches, (PCA-LDA, SVMs and Random Forests) are developed and tested on their ability to classify the cellular subtype in extracted peripheral blood mononuclear cells (T-cell lymphocytes from myeloid cells), and are evaluated in terms of their respective classification performance. A strategy for optimisation of each of the classification algorithm is presented …

The Utility Of Deformable Image Registration For Small Artery Visualisation In Contrast-Enhanced Whole Body Mr Angiography, Daniel Foley, Jacinta Browne, Xiahai Zhuang, Barry Sheane, Dearbhail O'Driscoll, Daniel Cannon, Niall Sheehy, James Meaney, Andrew Fagan

Articles

Purpose;

An investigation was carried out into the effect of three image registration techniques on the diagnostic image quality of contrast-enhanced magnetic resonance angiography (CE-MRA) images.

Methods Whole-body CE-MRA data from the lower legs of 27 patients recruited onto a study of asymptomatic atherosclerosis were processed using three deformable image registration algorithms. The resultant diagnostic image quality was evaluated qualitatively in a clinical evaluation by four expert observers, and quantitatively by measuring contrast-to-noise ratios and volumes of blood vessels, and assessing the techniques’ ability to correct for varying degrees of motion.

Results The first registration algorithm (‘AIR’) introduced significant stenosis-mimicking …

Breast Cancer Detection Using Interferometric Music: Experimental And Numerical Assessment, Giuseppe Ruvio, Raffaele Solimene, Antonio Cuccaro, Domenico Gaetano, Jacinta Browne, Max Ammann

Articles

Purpose: In microwave breast cancer detection it is often beneficial to arrange sensors in close proximity to the breast. The resulting coupling generally changes the antenna response. As an a priori characterization of the radio-frequency system becomes difficult, this can lead to severe degradation of the detection efficacy. The purpose of this paper is to demonstrate advantages of adopting an Interferometric Multiple Signal Classification (I-MUSIC) approach due to its limited dependence 25 from a priori information on the antenna. The performance of I-MUSIC detection was measured in terms of Signal-to-Clutter Ratio (SCR), Signal-to-Mean Ratio (SMR) and Spatial Displacement (SD) and …

Experimental Microwave Breast Cancer Detection With Oil-On-Gelatin Phantom, Giuseppe Ruvio, Raffaele Solimene, Antonio Cuccaro, Jacinta Browne, Domenico Gaetano, Max Ammann

Conference Papers

Limitations of conventional X-ray mammography has induced a recent interest into radio frequency based diagnostic approaches in order to take advantage of the sharpest dielectric contrast between benign and malignant breast tissues in the microwave frequency range [1]. Dielectric contrast is notably higher than the radiographic density exploited by X-ray mammography. Consequences of the superior sensitivity of RF-based techniques can have a huge social and cost impact. By reducing the false response percent, a dramatic reduction of costs for the health system together with a diminution of traumatic experiences for the patients can be achieved. The antenna properties play a …

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 …

Characterisation Of The Colour Doppler "Twinkle" Artefact, Deirdre Cassidy, Andrew Fagan, Jacinta Browne

Articles

This investigation involved the development of a range of Colour Doppler Twinkle Artefact phantoms to characterise and quantify the “Twinkle” artefact which is often present when an irregular structure is encountered in the imaged field of view. The artefact occurs in both colour and power Doppler ultrasound imaging and manifests as a false depiction of colour velocity information in stationary soft tissue and therefore can cause significant misdiagnosis of areas of flow within the patient. It has been hypothesised that it is generated due to a strongly reflecting medium composed of individual reflectors and therefore becomes a clinical concern when …

Comparative Review Of Imaging Methods Used For Diagnosing Renal Artery Stenosis (Ras), Deirdre King, Carmel Moran, Jacinta Browne

Articles

This comparative review examines the efficacy of different imaging methods to detect and quantify renal artery stenosis (RAS). Detection of RAS is important because it can lead to renovascular hypertension which is the most common form of secondary hypertension. Furthermore, it is important that a RAS is detected as early as possible as it is a potentially correctable cause of hypertension.¹ If detected at an early stage RAS can potentially be treated using a minimally invasive drug treatment regimen rather than the more invasive percutaneous transluminal renal angioplasty without or with stent placement or surgery. Currently there are a …

Quantitative Reagent-Free Detection Of Fibrinogen Levels In Human Blood Plasma Using Raman Spectroscopy, Kelvin Poon, Fiona Lyng, Peter Knief, Orla L. Howe, Aidan Meade, James Curtin, Hugh Byrne, Joseph Vaughan

Articles

Fibrinogen assays are commonly used as part of clinical screening tests to investigate haemorrhagic states, for detection of disseminated intravascular coagulation and as a predictor of a variety of cardiovascular events. The Clauss assay, which measures thrombin clotting time, is the most commonly used method for measuring fibrinogen levels. Nevertheless, inconsistencies are present in inter-manufacturer reagent sources, calibration standards and methodologies. Automated coagulation analysers, which measure changes in optical density during the prothrombin time (PT-Fg), have found use in many hospitals. However, the PT-Fg method is found to give falsely elevated values due to varying choices of calibrants, reagents and …

Understanding The Molecular Information Contained In Principal Component Analysis Of Vibrational Spectra Of Biological Systems, Franck Bonnier, Hugh Byrne

Articles

K-means clustering followed by Principal Component Analysis (PCA) is employed to analyse Raman spectroscopic maps of single biological cells. K-means clustering successfully identifies regions of cellular cytoplasm, nucleus and nucleoli, but the mean spectra do not differentiate their biochemical composition. The loadings of the principal components identified by PCA shed further light on the spectral basis for differentiation but they are complex and, as the number of spectra per cluster is imbalanced, particularly in the case of the nucleoli, the loadings under-represent the basis for differentiation of some cellular regions. Analysis of pure bio-molecules, both structurally and spectrally distinct, in …

Macular Pigment: Practical Implications For Optometric Practice In Preventative Health Care And Visual Performance Enhancement, Grainne Scanlon

Masters

The macula is a specialised part of the retina responsible for detailed central and colour vision. The carotenoids, lutein, zeaxanthin and meso-zeaxanthin are uniquely concentrated in the inner and central layers of the primate macula, where they are known as macular pigment (MP). It has been shown that MP is entirely of dietary origin and that lutein and zeaxanthin levels in serum, diet and retina correlate. Age-Related Macular Degeneration (AMD) is a disease of the macula and results in loss of central vision. MP, because of its optical filtration and antioxidant properties, may have an important role in the prevention …

Comparative Imaging Study In Ultrasound, Mri, Ct And Dsa Using A Multi-Modality Renal Artery Phantom, Deirdre King, Andrew Fagan, Carmel Moran, Jacinta Browne

Articles

A range of anatomically realistic multimodality renal artery phantoms consisting of vessels with varying degrees of stenosis was developed and evaluated using four imaging techniques currently used to detect renal artery stenosis (RAS). The spatial resolution required to visualize vascular geometry and the velocity detection performance required to adequately characterize blood flow in patients suffering from RAS are currently ill-defined, with the result that no one imaging modality has emerged as a gold standard technique for screening for this disease.

Methods:

The phantoms, which contained a range of stenosis values (0%, 30%, 50%, 70%, and 85%), were designed for use …

Evaluation Of The Potential Of Raman Microspectroscopy For Prediction Of Chemotherapeutic Response To Cisplatin In Lung Adenocarcinoma, Haq Nawaz, Franck Bonnier, Peter Knief, Orla Howe, Fiona Lyng, Aidan Meade, Hugh Byrne

Articles

The study of the interaction of anticancer drugs with mammalian cells in vitro is important to elucidate the mechanisms of action of the drug on its biological targets. In this context, Raman spectroscopy is a potential candidate for high throughput, noninvasive analysis. To explore this potential, the interaction of cis-Diamminedichloroplatinum (II) (Cisplatin) with a human lung adenocarcinoma cell line (A549) was investigated using Raman microspectroscopy. The results were correlated with parallel measurements from the MTT cytotoxicity assay, which yielded an IC50 value of 1.2±0.2 μM. To further confirm the spectral results, Raman spectra were also acquired from DNA extracted from …

Metal-Induced Bystander Effects: Mechanism And Implicationis., Nicola Cogan

Doctoral

The radiation-induced bystander effect is a phenomenon known to occur post irradiation, characterised by the induction of biological effects in unirradiated cells as a result of receiving signals from irradiated cells or their culture medium. Chemicalinduced bystander effects are poorly characterised and there are no reports of a bystander effect induced by metals. Heavy metals and in particular chromium (VI) are known to cause persistent genomic instability. For the first time, this study provides evidence that a short, low-dose exposure of human fibroblasts to chromium (VI) causes a bystander effect in human fibroblasts that persists for at least thirty days …

Swcnt Suppress Inflammatory Mediator Responses In Human Lung Epithelium In Vitro, Eva Herzog, Hugh Byrne, Maria Davoren, Anke-Gabriele Lenz, Albert Duschl, Gertie Janneke Oostingh

Articles

Single walled carbon nanotubes have gained enormous popularity due to a variety of potential applications which will ultimately lead to increased human and environmental exposure to these nanoparticles. This study was carried out in order to evaluate the inflammatory response of immortalised and primary human lung epithelial cells (A549 and NHBE) to single walled carbon nanotube samples (SWCNT). Special focus was placed on the mediating role of lung surfactant on particle toxicity. The toxicity of SWCNT dispersed in cell culture medium was compared to that of nanotubes dispersed in dipalmitoylphosphatidylcholine (DPPC, the main component of lung lining fluid). Exposure was …

Spectroscopic Study Of The Dimerisation Process Of Iron Protoporphyrin Ix, K. Dziedzic-Kocurek, Hugh Byrne, A. Swiderski, J. Stanek

Articles

The commercial protoporphyrin IX, iron-ferriprotoporphyrin IX-chloride and synthesized iron porphyrin μ-oxo-dimers were examined by UV/vis absorption and fluorescence, Fourier transformed infrared spectroscopy, resonance Raman, X-ray absorption, Mössbauer spectroscopy and SQUID. The evidence of Fe-O-Fe antiferromagnetic coupling concluded from SQUID and Mössbauer in the case of samples containing dimerized forms confirmed the presence of the oxo-bridges. In this paper the results of UV/vis, fluorescence, Fourier transform infrared FTIR and Raman spectroscopies are reported and discussed. The study is based on the comparison of the free-base protoporphyrin IX, Fe-PPIX-Cl and the synthesized dimerized specimen. The vibrational modes in two energy regions i.e. …

Development Of A Novel Modular Mid-Infrared Sensor For The In-Situ Detection Of The Btex Compounds In Water, Raymond Mccue

Doctoral

This research thesis describes the design, construction and testing of a novel modular mid-infrared fibre optic sensing system for the detection of hydrocarbons in water. It is the adverse effects to these hydrocarbon pollutants on flora and fauna that has led to the development of sensing systems for their detection and quantification. The key sensor design feature, its modularity, utilises simple low cost commercially available optical components, which are inherently suited to construct compact rugged sensing systems to perform in-situ measurements which are ideally preferred for environmental sensing. The various laboratory-based prototypes constructed weee calibrated for two target analytes over …

Colour Doppler Spatial Resolution Performance Testing, Jacinta Browne, Ian Brown, Peter Hoskins, Amanda Watson, Alex Elliott

Articles

No abstract available

Studies In Cardiac Defribrillation, Eamonn Mckenna

Masters

Cardiac disease resulting in cardiac arrest is one of the most common fatal diseases in the developed world. Defibrillation is the medical procedure used to correct the irregular cardiac rhythm (ventricular fibrillation (VF)) which is very often detected in patients suffering cardiac arrest. In the work reported in this thesis, two aspects of the defibrillation of patients in VF were examined. The first area of investigation concentrated on the use of multiple defibrillation electrodes to be used during closed chest (transthoracic) defibrillation. This study was carried out in a saline bath and centred on “focusing” electric fields from perimeter electrodes …