MALDI-mass spectrometry imaging (MALDI-MSI) is a technique that allows proteomic information, tha... more MALDI-mass spectrometry imaging (MALDI-MSI) is a technique that allows proteomic information, that is, the spatial distribution and identification of proteins, to be obtained directly from tissue sections. The use of in situ enzymatic digestion as a sample pretreatment prior to MALDI-MSI analysis has been found to be useful for retrieving protein identification directly from formalin-fixed, paraffin-embedded (ffpe) tissue sections. Here, an improved method for the study of the distribution and the identification of peptides obtained after in situ digestion of fppe pancreatic tumor tissue sections by using MALDI-mass spectrometry imaging coupled with ion mobility separation (IMS) is described. MALDI-IMS-MS images of peptide obtained from pancreatic tumor tissue sections allowed the localization of tumor regions within the tissue section, while minimizing the peak interferences which were observed with conventional MALDI-TOF MSI. The use of ion mobility separation coupled with MALDI-MSI improved the selectivity and specificity of the method and, hence, enabled both the localization and in situ identification of glucose regulated protein 78 kDa (Grp78), a tumor biomarker, within pancreatic tumor tissue sections. These findings were validated using immunohistochemical staining.
People interested in the research are advised to contact the author for the final version of the ... more People interested in the research are advised to contact the author for the final version of the publication, or visit the DOI to the publisher's website. • The final author version and the galley proof are versions of the publication after peer review. • The final published version features the final layout of the paper including the volume, issue and page numbers. Link to publication General rights Copyright and moral rights for the publications made accessible in the public portal are retained by the authors and/or other copyright owners and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights. • Users may download and print one copy of any publication from the public portal for the purpose of private study or research. • You may not further distribute the material or use it for any profit-making activity or commercial gain • You may freely distribute the URL identifying the publication in the public portal. If the publication is distributed under the terms of Article 25fa of the Dutch Copyright Act, indicated by the "Taverne" license above, please follow below link for the End User Agreement:
encountered in a forensic context, have not been as widely studied. An investigation into the mos... more encountered in a forensic context, have not been as widely studied. An investigation into the most suitable analytical techniques for the characterisation of bone samples exhumed from a variety of burial environments has been carried out. An understanding of the behaviour of bone exposed to different conditions will provide forensic practitioners with additional tools to characterise bones. Bone samples were produced using pig bone as a model for human bone. The environmental factors were controlled in closed containers in the laboratory. Thermal analysis techniques, including differential thermal analysis and thermogravimetric analysis, mass spectrometry, can provide information regarding the organic and mineral content of bones. Studies at the University of Technology, Sydney (UTS) have demonstrated the potential of thermal analysis as a technique for characterising changes in bones. In addition, pyrolysis gas chromatography mass spectrometry has been applied in a study of pig bones subjected to controlled burials. The study has demonstrated a promising correlation between the post-mortem age of the bone and specific marker peaks in the data. Scanning electron microscopy has provided information regarding the morphology of the bone structure. X-ray diffraction was used to characterise the mineral content of bones. In addition, infrared spectroscopy has been employed to provide important information regarding the chemical structure of bone samples.
Over the past seven years Matrix Assisted Laser Desorption Ionisation Mass Spectrometry Profiling... more Over the past seven years Matrix Assisted Laser Desorption Ionisation Mass Spectrometry Profiling (MALDI MSP) and Imaging (MALDI MSI) have proven to be feasible tools for the detection of blood and its provenance in stains and fingermarks. However, whilst this capability as a confirmatory test addresses the primary questions at the scene of a violent crime, additional intelligence recoverable from blood can also prove important for investigations. A DNA profile is the most obvious and important example of such intelligence; however, it is not always suitable for identification purposes, depending on quantity, age and environmental conditions. Proteins are much more stable and determining the presence of haemoglobin variants in blood recovered at a crime scene may provide associative and possibly corroborating evidence on the presence of an individual at a particular location. This evidence gains more incriminatory value, the lower the incidence of the variant in a certain geographical area or population and may contribute to narrowing down the pool of suspects. In this study, a MALDI based mass spectrometric method has been developed and tested on six haemoglobin variants for their fast and reliable identification and mapping in blood fingermarks. † Electronic supplementary information (ESI) available. See
Eight years of intensive research have demonstrated that Matrix-Assisted Laser Desorption/Ionisat... more Eight years of intensive research have demonstrated that Matrix-Assisted Laser Desorption/Ionisation Mass Spectrometry Profiling and Imaging (MALDI-MSI and MSP) are powerful tools to gather intelligence around a suspect lifestyle, directly from the identifying ridges of a latent fingermark. In the past three years, many efforts have been invested into translating laboratory methodologies to the field; this was undertaken by devising protocols either for (a) enabling initial fingermark visualisation, such as through the Dry-Wet method, recovery and subsequent MALDI MS based analysis, or for (b) rendering the MS methodologies compatible with the prior application of commonly employed fingermark enhancement techniques (FET). In the present work a major point of interest concerned the sample treatment of FET visualised-lifted fingermarks and the subsequent MS performance of primary tape lifted fingermarks ("primary lifts") versus secondary tape lifted fingermarks (recovery from the surface a second time following the initial primary lift). This was necessary since it may not always be possible to obtain primary lifts of marks visualised at crime scenes for remote MALDI-MSP and MSI. The work illustrated here has provided methodological insights into establishing how to best treat a few types of developed marks in preparation for MALDI-MSI when presented as both secondary and primary lifts; it was demonstrated, as expected, that primary lifts generally yield much higher quality chemical/physical information and are therefore crucial to maximise chances of suspect identification and of retrieval of chemical intelligence. When analysing secondary lifted marks that have been initially developed using aluminium or carbon powders, any of the trialled sample preparation methodologies can be employed except the Dry-Wet method. In the case of TiO 2 powder developed marks, the best ridge coverage was achieved by re-enhancing the mark using the initial powder and spray-coating with MALDI matrix. Primary lifts of fingermarks contaminated with an exogenous substance (used as a reference model) yielded the best ridge detail quality whilst for secondary lifts of natural marks pre-enhanced with aluminium powder, significantly greater intensity of the ion image was observed for the sections subjected to either no further enhancement or re-enhancement using aluminium powder.
International Journal for Ion Mobility Spectrometry, Feb 27, 2013
Matrix assisted laser desorption ionisation ion mobility separation mass spectrometry imaging (MA... more Matrix assisted laser desorption ionisation ion mobility separation mass spectrometry imaging (MALDI-IMS-MSI) has been employed to image the distribution of proteins in ex-vivo human skin. Using a "bottom-up" proteomics approach employing "on-tissue" digestion the distribution of abundant skin proteins; collagen, keratin, decorin and serum albumin could be mapped. Images have been recorded at 150 and 30 μm spatial resolution. Multivariate statistical analysis of the data has been employed to associate specific proteins with layers of the skin. The improved specificity given by the use of ion mobility separation in mass spectrometric imaging has been demonstrated by separation of peptide ions from phospholipids.
In the study presented here, four examples of crime scene fingermarks analysed by Matrix Assisted... more In the study presented here, four examples of crime scene fingermarks analysed by Matrix Assisted Laser Desorption Ionisation Mass Spectrometry Imaging (MALDI MSI), in collaboration with the local Police Force, are reported. These marks are associated to high profile crimes such as drug dealing, murder and harassment. Following the application of forensic enhancement techniques by the CSI or the crime lab, one of the marks could be directly analysed on the surface of deposition and the others were analysed as primary lifts. In one case, no physical or molecular information was obtained whereas in two cases both ridge detail (albeit not useful for identification purposes) and molecular information could be retrieved. In one case, the intelligence gathered around the suspect's state of mind is, to date the most accomplished demonstration of the benefits and the operational feasibility MALDI MS based methods. These four casework examples are to be considered a first insight into the limitations, benefits and feasibility of MALDI MS based methods in the field; due to the extreme variability in the state of the mark, comprehensive evaluation of these aspects can only be undertaken upon the analysis of a large number of crime scene marks. However, this work does represent a significant advancement, compared to previous published work, as it demonstrates operational feasibility, with different levels of success, despite uncontrollable, unknown and unaccountable environmental and donor variability.
The determination of the presence of blood in fingermarks constitutes important intelligence in a... more The determination of the presence of blood in fingermarks constitutes important intelligence in a criminal investigation as it helps to reconstruct the events that have taken place at a scene of crime. Various methodologies have been reported and are currently employed for the detection of the presence of blood including optical, spectroscopic and chemical development approaches. However, most methods only give an indication that blood may be present and, therefore, these methods are described as presumptive tests. Here we show the use of Matrix-Assisted Laser Desorption Ionisation Mass Spectrometry Profiling and Imaging (MALDI MSP and MALDI MSI) for the determination of the presence of blood in fingermarks by specifically detecting the molecules of haem and haemoglobin through their mass-to-charge ratios. Furthermore, preliminary experiments are shown which demonstrate that this technology is compatible with other methods currently employed for enhancing fingermarks in blood (or contaminated by blood). The application of the developed protocols to a crime scene blood trace, demonstrates the feasibility of using this technology in routine casework. These findings open up a new line of research for the development of robust MALDI MSP and MALDI MSI protocols for the detection and chemical imaging of bloodied marks.
Journal of the American Society for Mass Spectrometry, 2009
A new analytical approach using matrix-assisted laser desorption/ionization mass spectrometry ima... more A new analytical approach using matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) for the study of honeybee venom is shown. In vitro and in vivo models simulating the bee sting have been developed using live honeybees and, as the envenomation sites, pig ears and rat legs; MALDI MSI has been used to map, over time, the diffusion and distribution of three venom allergens (Api m 1, Api m 4, and Api m 6) and two venom toxins (apamine and mast cell degranulating peptide). In conjunction with other classical biochemical techniques and high resolution mass spectrometry (HRMS), structural data have been obtained that contribute to current understanding of honeybee venom composition. Initial data have also been obtained demonstrating the feasibility of mapping the organism's response to the sting. The opportunity to monitor venom diffusion and the organism's response at the same time might open new pathways for in vivo preclinical studies in designing and testing new venom immunotherapy (VIT).
Clioquinol (CQ) is a &amp... more Clioquinol (CQ) is a "metal protein attenuating compound" that crosses the blood-brain barrier and binds, with high affinity, copper(II) and zinc(II), two metal ions critically involved in amyloid-beta aggregation and toxicity. CQ was recently proposed for the treatment of Alzheimer's disease, but controversial data have been reported so far concerning its real therapeutic advantages. We describe here results of chronic CQ treatment in the TgCRND8 mouse model of Alzheimer's disease. Remarkably, based on classical behavioral tests, CQ treatment was found to reverse, to a large extent, the working memory impairments that are characteristic of this mouse model. Pairwise, a significant reduction of amyloid-beta plaque burden, both in the cortex and in the hippocampus, was detected as well as an attenuation of astrogliosis. MALDI Mass Spectrometry Imaging technique revealed a specific localization of CQ in the above mentioned brain areas. Modest but significant effects on the absolute and relative brain concentrations of the three most important biometals (i.e., copper, zinc, and iron) were highlighted following CQ treatment. The pharmacological and mechanistic implications of the above findings are thoroughly discussed.
This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY
Breast cancer is a global health issue affecting 2.3 million women per year, causing death in ove... more Breast cancer is a global health issue affecting 2.3 million women per year, causing death in over 600,000. Mammography (and biopsy) is the gold standard for screening and diagnosis. Whilst effective, this test exposes individuals to radiation, has limitations to its sensitivity and specificity and may cause moderate to severe discomfort. Some women may also find this test culturally unacceptable. This proof-of-concept study, combining bottom-up proteomics with Matrix Assisted Laser Desorption Ionisation Mass Spectrometry (MALDI MS) detection, explores the potential for a non-invasive technique for the early detection of breast cancer from fingertip smears. A cohort of 15 women with either benign breast disease (n = 5), early breast cancer (n = 5) or metastatic breast cancer (n = 5) were recruited from a single UK breast unit. Fingertips smears were taken from each patient and from each of the ten digits, either at the time of diagnosis or, for metastatic patients, during active treatment. A number of statistical analyses and machine learning approaches were investigated and applied to the resulting mass spectral dataset. The highest performing predictive method, a 3-class Multilayer Perceptron neural network, yielded an accuracy score of 97.8% when categorising unseen MALDI MS spectra as either the benign, early or metastatic cancer classes. These findings support the need for further research into the use of sweat deposits (in the form of fingertip smears or fingerprints) for non-invasive screening of breast cancer. Breast cancer is a global health issue. In the UK 55,920 new cases of breast cancer were diagnosed annually (2016-2018), making it the most common cancer in British women 1. The World Health Organisation (WHO) has reported that, in 2020, 2.3 million women were diagnosed with breast cancer, and approximately 30% of them had a fatal outcome, globally 2. Furthermore, according to the WHO, breast cancer is the world's most prevalent malignant neoplasm, given that, by the end of 2020, 7.8 million living women were registered to have received a breast cancer diagnosis in the past 5 years 2. Rates of, and mortality from breast cancer are rising globally due to changes in population age structure and lifestyle issues. There is significant variation in outcomes, both between and within nations, due in part to variation in access to mammography, the mainstay of breast cancer screening and diagnosis 3 and variation in access to systemic therapies. Although mammography has facilitated earlier diagnosis and consequent improved outcomes, it is associated with exposure to radiation and physical discomfort. Mammography may be painful for many women as it requires breast compression between compression plates to enhance sensitivity. Additionally, some women find exposure of their breasts for mammography culturally unacceptable and decline screening, or delay it, even when they experience symptoms. In addition, about 10% of cancers are not visible on mammography due to high breast density 4-6 or with certain biological subtypes such as lobular cancer 7. In addition, in women under the age of 40, mammography has a very low sensitivity due to high breast density in this age group. These limitations to the sensitivity of mammography may lead to delayed diagnosis which results in increased mortality rates and treatment morbidity (greater need for mastectomy, axillary clearance, chemotherapy, radiotherapy). In addition, the cost and resource issues of population screening with mammography are significant.
For decades, the analysis of fingerprints (FP) has been used as the primary biometric mean of hum... more For decades, the analysis of fingerprints (FP) has been used as the primary biometric mean of human identification. In parallel, the chemical analysis of latent fingermark (LFM) with particular reference to "touch chemistry", offers additional intelligence to forensic examination; as such, continuous improvements in the versatility and sensitivity of detection of the molecular make up of FM is of obvious importance. In this light, we propose a facile approach to synthesize magnetic carbon nanoparticles (MCNPs) from candle soot for extraction and detection of endogenous and exogenous FM components. Initially, endogenous components of FM including fatty acids (FAs), squalene and triacylgycerols (TAGs), were successfully extracted and detected using the developed MCNPs and surface assisted laser desorption/ionization-mass spectrometry (SALDI-MS). Furthermore, the MCNPs enabled the detection of exogenous substances including antihistamine, β-blocker, antibiotic drugs and lead in contaminated FMs, whilst providing characteristic and unique fragmentation patterns for each drugs in the FM. The influence of environmental factors such as temperature on the stability of the exogenous substances in FM was investigated by exposing the contaminated FM to different temperatures over 24 hours, the findings revealed the drugs' instability at high temperatures and undergo different degrees of degradation whereas lead was more resilient to thermal stress. The detection of triprolidine, metoprolol and sulfamethoxazole from pharmaceutical tablets in FM was successfully achieved by gently touching the tablet powder. The limit of detection (LOD) values of the drugs in the tablet forms were 50, 200 and 750 ng mL-1 for triprolidine, metoprolol and sulfamethoxazole, respectively and their recovery rates were 91.17% for triprolidine, 94.67% for metoprolol and 120.86% for sulfamethoxazole. Finally, to create a genuine casework scenario, contaminated FM was deposited on glass and metal surfaces then subjected to extraction using MCNPs and magnet without compromising the FM impression. Substrate control experiments revealed that the glass surface exhibiting some background signal, however, they did not interfere with the analysis and satisfactory extraction efficiency of endogenous and exogenous components of the FM on both surfaces was obtained using the MCNPs. Overall, this study proves the capability of MCNPs as new SALDI-MS substrate for both extraction and detection of FM components gathering more information pertaining to the donor's lifestyle.
ABSTRACT no The identification of proteins involved in tumour progression or which permit enhance... more ABSTRACT no The identification of proteins involved in tumour progression or which permit enhanced or novel therapeutic targeting is essential for cancer research. Direct MALDI analysis of tissue sections is rapidly demonstrating its potential for protein imaging and profiling in the investigation of a range of disease states including cancer. MALDI-mass spectrometry imaging (MALDI-MSI) has been used here for direct visualisation and in situ characterisation of proteins in breast tumour tissue section samples. Frozen MCF7 breast tumour xenograft and human formalin-fixed paraffin-embedded breast cancer tissue sections were used. An improved protocol for on-tissue trypsin digestion is described incorporating the use of a detergent, which increases the yield of tryptic peptides for both fresh frozen and formalin-fixed paraffin-embedded tumour tissue sections. A novel approach combining MALDI-MSI and ion mobility separation MALDI-tandem mass spectrometry imaging for improving the detection of low-abundance proteins that are difficult to detect by direct MALDI-MSI analysis is described. In situ protein identification was carried out directly from the tissue section by MALDI-MSI. Numerous protein signals were detected and some proteins including histone H3, H4 and Grp75 that were abundant in the tumour region were identified
MALDI-mass spectrometry imaging (MALDI-MSI) is a technique that allows proteomic information, tha... more MALDI-mass spectrometry imaging (MALDI-MSI) is a technique that allows proteomic information, that is, the spatial distribution and identification of proteins, to be obtained directly from tissue sections. The use of in situ enzymatic digestion as a sample pretreatment prior to MALDI-MSI analysis has been found to be useful for retrieving protein identification directly from formalin-fixed, paraffin-embedded (ffpe) tissue sections. Here, an improved method for the study of the distribution and the identification of peptides obtained after in situ digestion of fppe pancreatic tumor tissue sections by using MALDI-mass spectrometry imaging coupled with ion mobility separation (IMS) is described. MALDI-IMS-MS images of peptide obtained from pancreatic tumor tissue sections allowed the localization of tumor regions within the tissue section, while minimizing the peak interferences which were observed with conventional MALDI-TOF MSI. The use of ion mobility separation coupled with MALDI-MSI improved the selectivity and specificity of the method and, hence, enabled both the localization and in situ identification of glucose regulated protein 78 kDa (Grp78), a tumor biomarker, within pancreatic tumor tissue sections. These findings were validated using immunohistochemical staining.
People interested in the research are advised to contact the author for the final version of the ... more People interested in the research are advised to contact the author for the final version of the publication, or visit the DOI to the publisher's website. • The final author version and the galley proof are versions of the publication after peer review. • The final published version features the final layout of the paper including the volume, issue and page numbers. Link to publication General rights Copyright and moral rights for the publications made accessible in the public portal are retained by the authors and/or other copyright owners and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights. • Users may download and print one copy of any publication from the public portal for the purpose of private study or research. • You may not further distribute the material or use it for any profit-making activity or commercial gain • You may freely distribute the URL identifying the publication in the public portal. If the publication is distributed under the terms of Article 25fa of the Dutch Copyright Act, indicated by the "Taverne" license above, please follow below link for the End User Agreement:
encountered in a forensic context, have not been as widely studied. An investigation into the mos... more encountered in a forensic context, have not been as widely studied. An investigation into the most suitable analytical techniques for the characterisation of bone samples exhumed from a variety of burial environments has been carried out. An understanding of the behaviour of bone exposed to different conditions will provide forensic practitioners with additional tools to characterise bones. Bone samples were produced using pig bone as a model for human bone. The environmental factors were controlled in closed containers in the laboratory. Thermal analysis techniques, including differential thermal analysis and thermogravimetric analysis, mass spectrometry, can provide information regarding the organic and mineral content of bones. Studies at the University of Technology, Sydney (UTS) have demonstrated the potential of thermal analysis as a technique for characterising changes in bones. In addition, pyrolysis gas chromatography mass spectrometry has been applied in a study of pig bones subjected to controlled burials. The study has demonstrated a promising correlation between the post-mortem age of the bone and specific marker peaks in the data. Scanning electron microscopy has provided information regarding the morphology of the bone structure. X-ray diffraction was used to characterise the mineral content of bones. In addition, infrared spectroscopy has been employed to provide important information regarding the chemical structure of bone samples.
Over the past seven years Matrix Assisted Laser Desorption Ionisation Mass Spectrometry Profiling... more Over the past seven years Matrix Assisted Laser Desorption Ionisation Mass Spectrometry Profiling (MALDI MSP) and Imaging (MALDI MSI) have proven to be feasible tools for the detection of blood and its provenance in stains and fingermarks. However, whilst this capability as a confirmatory test addresses the primary questions at the scene of a violent crime, additional intelligence recoverable from blood can also prove important for investigations. A DNA profile is the most obvious and important example of such intelligence; however, it is not always suitable for identification purposes, depending on quantity, age and environmental conditions. Proteins are much more stable and determining the presence of haemoglobin variants in blood recovered at a crime scene may provide associative and possibly corroborating evidence on the presence of an individual at a particular location. This evidence gains more incriminatory value, the lower the incidence of the variant in a certain geographical area or population and may contribute to narrowing down the pool of suspects. In this study, a MALDI based mass spectrometric method has been developed and tested on six haemoglobin variants for their fast and reliable identification and mapping in blood fingermarks. † Electronic supplementary information (ESI) available. See
Eight years of intensive research have demonstrated that Matrix-Assisted Laser Desorption/Ionisat... more Eight years of intensive research have demonstrated that Matrix-Assisted Laser Desorption/Ionisation Mass Spectrometry Profiling and Imaging (MALDI-MSI and MSP) are powerful tools to gather intelligence around a suspect lifestyle, directly from the identifying ridges of a latent fingermark. In the past three years, many efforts have been invested into translating laboratory methodologies to the field; this was undertaken by devising protocols either for (a) enabling initial fingermark visualisation, such as through the Dry-Wet method, recovery and subsequent MALDI MS based analysis, or for (b) rendering the MS methodologies compatible with the prior application of commonly employed fingermark enhancement techniques (FET). In the present work a major point of interest concerned the sample treatment of FET visualised-lifted fingermarks and the subsequent MS performance of primary tape lifted fingermarks ("primary lifts") versus secondary tape lifted fingermarks (recovery from the surface a second time following the initial primary lift). This was necessary since it may not always be possible to obtain primary lifts of marks visualised at crime scenes for remote MALDI-MSP and MSI. The work illustrated here has provided methodological insights into establishing how to best treat a few types of developed marks in preparation for MALDI-MSI when presented as both secondary and primary lifts; it was demonstrated, as expected, that primary lifts generally yield much higher quality chemical/physical information and are therefore crucial to maximise chances of suspect identification and of retrieval of chemical intelligence. When analysing secondary lifted marks that have been initially developed using aluminium or carbon powders, any of the trialled sample preparation methodologies can be employed except the Dry-Wet method. In the case of TiO 2 powder developed marks, the best ridge coverage was achieved by re-enhancing the mark using the initial powder and spray-coating with MALDI matrix. Primary lifts of fingermarks contaminated with an exogenous substance (used as a reference model) yielded the best ridge detail quality whilst for secondary lifts of natural marks pre-enhanced with aluminium powder, significantly greater intensity of the ion image was observed for the sections subjected to either no further enhancement or re-enhancement using aluminium powder.
International Journal for Ion Mobility Spectrometry, Feb 27, 2013
Matrix assisted laser desorption ionisation ion mobility separation mass spectrometry imaging (MA... more Matrix assisted laser desorption ionisation ion mobility separation mass spectrometry imaging (MALDI-IMS-MSI) has been employed to image the distribution of proteins in ex-vivo human skin. Using a "bottom-up" proteomics approach employing "on-tissue" digestion the distribution of abundant skin proteins; collagen, keratin, decorin and serum albumin could be mapped. Images have been recorded at 150 and 30 μm spatial resolution. Multivariate statistical analysis of the data has been employed to associate specific proteins with layers of the skin. The improved specificity given by the use of ion mobility separation in mass spectrometric imaging has been demonstrated by separation of peptide ions from phospholipids.
In the study presented here, four examples of crime scene fingermarks analysed by Matrix Assisted... more In the study presented here, four examples of crime scene fingermarks analysed by Matrix Assisted Laser Desorption Ionisation Mass Spectrometry Imaging (MALDI MSI), in collaboration with the local Police Force, are reported. These marks are associated to high profile crimes such as drug dealing, murder and harassment. Following the application of forensic enhancement techniques by the CSI or the crime lab, one of the marks could be directly analysed on the surface of deposition and the others were analysed as primary lifts. In one case, no physical or molecular information was obtained whereas in two cases both ridge detail (albeit not useful for identification purposes) and molecular information could be retrieved. In one case, the intelligence gathered around the suspect's state of mind is, to date the most accomplished demonstration of the benefits and the operational feasibility MALDI MS based methods. These four casework examples are to be considered a first insight into the limitations, benefits and feasibility of MALDI MS based methods in the field; due to the extreme variability in the state of the mark, comprehensive evaluation of these aspects can only be undertaken upon the analysis of a large number of crime scene marks. However, this work does represent a significant advancement, compared to previous published work, as it demonstrates operational feasibility, with different levels of success, despite uncontrollable, unknown and unaccountable environmental and donor variability.
The determination of the presence of blood in fingermarks constitutes important intelligence in a... more The determination of the presence of blood in fingermarks constitutes important intelligence in a criminal investigation as it helps to reconstruct the events that have taken place at a scene of crime. Various methodologies have been reported and are currently employed for the detection of the presence of blood including optical, spectroscopic and chemical development approaches. However, most methods only give an indication that blood may be present and, therefore, these methods are described as presumptive tests. Here we show the use of Matrix-Assisted Laser Desorption Ionisation Mass Spectrometry Profiling and Imaging (MALDI MSP and MALDI MSI) for the determination of the presence of blood in fingermarks by specifically detecting the molecules of haem and haemoglobin through their mass-to-charge ratios. Furthermore, preliminary experiments are shown which demonstrate that this technology is compatible with other methods currently employed for enhancing fingermarks in blood (or contaminated by blood). The application of the developed protocols to a crime scene blood trace, demonstrates the feasibility of using this technology in routine casework. These findings open up a new line of research for the development of robust MALDI MSP and MALDI MSI protocols for the detection and chemical imaging of bloodied marks.
Journal of the American Society for Mass Spectrometry, 2009
A new analytical approach using matrix-assisted laser desorption/ionization mass spectrometry ima... more A new analytical approach using matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) for the study of honeybee venom is shown. In vitro and in vivo models simulating the bee sting have been developed using live honeybees and, as the envenomation sites, pig ears and rat legs; MALDI MSI has been used to map, over time, the diffusion and distribution of three venom allergens (Api m 1, Api m 4, and Api m 6) and two venom toxins (apamine and mast cell degranulating peptide). In conjunction with other classical biochemical techniques and high resolution mass spectrometry (HRMS), structural data have been obtained that contribute to current understanding of honeybee venom composition. Initial data have also been obtained demonstrating the feasibility of mapping the organism's response to the sting. The opportunity to monitor venom diffusion and the organism's response at the same time might open new pathways for in vivo preclinical studies in designing and testing new venom immunotherapy (VIT).
Clioquinol (CQ) is a &amp... more Clioquinol (CQ) is a "metal protein attenuating compound" that crosses the blood-brain barrier and binds, with high affinity, copper(II) and zinc(II), two metal ions critically involved in amyloid-beta aggregation and toxicity. CQ was recently proposed for the treatment of Alzheimer's disease, but controversial data have been reported so far concerning its real therapeutic advantages. We describe here results of chronic CQ treatment in the TgCRND8 mouse model of Alzheimer's disease. Remarkably, based on classical behavioral tests, CQ treatment was found to reverse, to a large extent, the working memory impairments that are characteristic of this mouse model. Pairwise, a significant reduction of amyloid-beta plaque burden, both in the cortex and in the hippocampus, was detected as well as an attenuation of astrogliosis. MALDI Mass Spectrometry Imaging technique revealed a specific localization of CQ in the above mentioned brain areas. Modest but significant effects on the absolute and relative brain concentrations of the three most important biometals (i.e., copper, zinc, and iron) were highlighted following CQ treatment. The pharmacological and mechanistic implications of the above findings are thoroughly discussed.
This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY
Breast cancer is a global health issue affecting 2.3 million women per year, causing death in ove... more Breast cancer is a global health issue affecting 2.3 million women per year, causing death in over 600,000. Mammography (and biopsy) is the gold standard for screening and diagnosis. Whilst effective, this test exposes individuals to radiation, has limitations to its sensitivity and specificity and may cause moderate to severe discomfort. Some women may also find this test culturally unacceptable. This proof-of-concept study, combining bottom-up proteomics with Matrix Assisted Laser Desorption Ionisation Mass Spectrometry (MALDI MS) detection, explores the potential for a non-invasive technique for the early detection of breast cancer from fingertip smears. A cohort of 15 women with either benign breast disease (n = 5), early breast cancer (n = 5) or metastatic breast cancer (n = 5) were recruited from a single UK breast unit. Fingertips smears were taken from each patient and from each of the ten digits, either at the time of diagnosis or, for metastatic patients, during active treatment. A number of statistical analyses and machine learning approaches were investigated and applied to the resulting mass spectral dataset. The highest performing predictive method, a 3-class Multilayer Perceptron neural network, yielded an accuracy score of 97.8% when categorising unseen MALDI MS spectra as either the benign, early or metastatic cancer classes. These findings support the need for further research into the use of sweat deposits (in the form of fingertip smears or fingerprints) for non-invasive screening of breast cancer. Breast cancer is a global health issue. In the UK 55,920 new cases of breast cancer were diagnosed annually (2016-2018), making it the most common cancer in British women 1. The World Health Organisation (WHO) has reported that, in 2020, 2.3 million women were diagnosed with breast cancer, and approximately 30% of them had a fatal outcome, globally 2. Furthermore, according to the WHO, breast cancer is the world's most prevalent malignant neoplasm, given that, by the end of 2020, 7.8 million living women were registered to have received a breast cancer diagnosis in the past 5 years 2. Rates of, and mortality from breast cancer are rising globally due to changes in population age structure and lifestyle issues. There is significant variation in outcomes, both between and within nations, due in part to variation in access to mammography, the mainstay of breast cancer screening and diagnosis 3 and variation in access to systemic therapies. Although mammography has facilitated earlier diagnosis and consequent improved outcomes, it is associated with exposure to radiation and physical discomfort. Mammography may be painful for many women as it requires breast compression between compression plates to enhance sensitivity. Additionally, some women find exposure of their breasts for mammography culturally unacceptable and decline screening, or delay it, even when they experience symptoms. In addition, about 10% of cancers are not visible on mammography due to high breast density 4-6 or with certain biological subtypes such as lobular cancer 7. In addition, in women under the age of 40, mammography has a very low sensitivity due to high breast density in this age group. These limitations to the sensitivity of mammography may lead to delayed diagnosis which results in increased mortality rates and treatment morbidity (greater need for mastectomy, axillary clearance, chemotherapy, radiotherapy). In addition, the cost and resource issues of population screening with mammography are significant.
For decades, the analysis of fingerprints (FP) has been used as the primary biometric mean of hum... more For decades, the analysis of fingerprints (FP) has been used as the primary biometric mean of human identification. In parallel, the chemical analysis of latent fingermark (LFM) with particular reference to "touch chemistry", offers additional intelligence to forensic examination; as such, continuous improvements in the versatility and sensitivity of detection of the molecular make up of FM is of obvious importance. In this light, we propose a facile approach to synthesize magnetic carbon nanoparticles (MCNPs) from candle soot for extraction and detection of endogenous and exogenous FM components. Initially, endogenous components of FM including fatty acids (FAs), squalene and triacylgycerols (TAGs), were successfully extracted and detected using the developed MCNPs and surface assisted laser desorption/ionization-mass spectrometry (SALDI-MS). Furthermore, the MCNPs enabled the detection of exogenous substances including antihistamine, β-blocker, antibiotic drugs and lead in contaminated FMs, whilst providing characteristic and unique fragmentation patterns for each drugs in the FM. The influence of environmental factors such as temperature on the stability of the exogenous substances in FM was investigated by exposing the contaminated FM to different temperatures over 24 hours, the findings revealed the drugs' instability at high temperatures and undergo different degrees of degradation whereas lead was more resilient to thermal stress. The detection of triprolidine, metoprolol and sulfamethoxazole from pharmaceutical tablets in FM was successfully achieved by gently touching the tablet powder. The limit of detection (LOD) values of the drugs in the tablet forms were 50, 200 and 750 ng mL-1 for triprolidine, metoprolol and sulfamethoxazole, respectively and their recovery rates were 91.17% for triprolidine, 94.67% for metoprolol and 120.86% for sulfamethoxazole. Finally, to create a genuine casework scenario, contaminated FM was deposited on glass and metal surfaces then subjected to extraction using MCNPs and magnet without compromising the FM impression. Substrate control experiments revealed that the glass surface exhibiting some background signal, however, they did not interfere with the analysis and satisfactory extraction efficiency of endogenous and exogenous components of the FM on both surfaces was obtained using the MCNPs. Overall, this study proves the capability of MCNPs as new SALDI-MS substrate for both extraction and detection of FM components gathering more information pertaining to the donor's lifestyle.
ABSTRACT no The identification of proteins involved in tumour progression or which permit enhance... more ABSTRACT no The identification of proteins involved in tumour progression or which permit enhanced or novel therapeutic targeting is essential for cancer research. Direct MALDI analysis of tissue sections is rapidly demonstrating its potential for protein imaging and profiling in the investigation of a range of disease states including cancer. MALDI-mass spectrometry imaging (MALDI-MSI) has been used here for direct visualisation and in situ characterisation of proteins in breast tumour tissue section samples. Frozen MCF7 breast tumour xenograft and human formalin-fixed paraffin-embedded breast cancer tissue sections were used. An improved protocol for on-tissue trypsin digestion is described incorporating the use of a detergent, which increases the yield of tryptic peptides for both fresh frozen and formalin-fixed paraffin-embedded tumour tissue sections. A novel approach combining MALDI-MSI and ion mobility separation MALDI-tandem mass spectrometry imaging for improving the detection of low-abundance proteins that are difficult to detect by direct MALDI-MSI analysis is described. In situ protein identification was carried out directly from the tissue section by MALDI-MSI. Numerous protein signals were detected and some proteins including histone H3, H4 and Grp75 that were abundant in the tumour region were identified
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