Mixtures of the two minerals huntite and hydromagnesite have been successfully used as a fire retardant additive in polymers for many years. The onset of decomposition of hydromagnesite is at a higher temperature than that of aluminium hydroxide but lower than that of magnesium hydroxide, the two most commonly used mineral fire retardants. This makes it an ideal addition to the range of materials available to polymer compounders for improving fire retardant properties. In comparison to the better known mineral fire retardants there has been little published research on the fire retardant properties of huntite and hydromagnesite. What has been published has often been commercially orientated and the limited quantity of scientific literature does not fully explain the fire retardant mechanism of these blends of minerals, often dismissing huntite as having no useful fire retardant action other than diluting the solid phase fuel. Standard thermal analysis techniques (thermal gravimetric analysis, differential scanning calorimetry, Fourier transform infra-red analysis) have been used to characterise the thermal decomposition of huntite and hydromagnesite from a source in Turkey. This has lead to an understanding of the decomposition mechanism of the minerals in terms of mass loss, enthalpy of decomposition, and evolved gases between room temperature and 1000°C. Hydromagnesite endothermically decomposes between about 220°C and 500°C, initially releasing water followed by carbon dioxide. The rate of heating and partial pressure of carbon dioxide in the atmosphere can influence the mechanism of carbon dioxide release. Huntite endothermically decomposes between about 450°C and 800°C releasing carbon dioxide in two stages. The use of the cone calorimeter to study the rate of heat release during combustion of ethylene vinyl acetate based polymer compounds has lead to an understanding of how both huntite and hydromagnesite affect the burning processes at different stages of the fire. By varying the ratio of the two minerals, hydromagnesite has been shown to increase the time to ignition and reduce the initial peak in rate of heat release, while huntite has been shown to reduce the rate of heat release later in the fire. It has been shown that huntite is far from being an inactive diluent filler. The endothermic decomposition of huntite in the later stages of the fire reduces the heat reaching underlying polymer and continues to dilute the flame with inert carbon dioxide. The platy huntite particles have been shown to align themselves in such a way that they can hinder the escape of volatiles from the decomposing polymer and also physically reinforce the inorganic ash residue.

A series of core and core-shell nanoparticles with superparamagnetic properties were synthesised and surface functionalised using three different amino-silanes by a chemical conjugation method. The functionalised nanoparticles were characterised and further modified by chemical conjugation with two different classes of bio-molecules; (i) enzymes and (ii) single stranded DNA primers. The resultant nanoparticles (nano-bio conjugates) were used for applications in (i) enzyme catalysis and (ii) bio-separation / bio-sensing. Magnetite and amorphous silica-coated core-shell nanoparticles were synthesised on both small (5 g) and large (20 g) scales and were characterised using transmission electron microscopy (TEM), X-ray diffraction (XRD), Brunauer-Emmett-Teller (BET) surface area measurement and vibrating sample magnetometry (VSM). Silica-coated core-shell nanoparticles were functionalised by silanisation with three different aminosilanes [3-aminopropyl tri-ethoxysilane (APTS), 3-aminopropyl di-ethoxymethylsilane (APDS) and 3-aminopropyl mono-ethoxydimethylsilane (APMS)] and two different methods: water (classical method) or a Tri-phasic Reverse Emulsion (TPRE) using toluene and a surfactant (Triton X-100). It was observed that the materials prepared using the TPRE method produced higher surface amine density values on average. The first application involved bio-catalysis where lipases [Pseudomonas Fluorescens lipase (PFL) and Candida Rugosa lipase (CRL)] were chemically conjugated (covalently linked) via glutaraldehyde-modification onto the amino-functionalised nanoparticles for applications such as: (i) hydrolysis of p-nitrophenyl palmitate to produce palmitic acid and p-nitrophenol (model reaction), (ii) transesterification of ethyl butyrate with n-butanol to produce butyl butyrate and (iii) partial and selective hydrolysis of cis-3,5-diacetoxy-1-cyclopentene to produce pharmaceutically important and expensive chiral intermediate molecules. Various reaction parameters such as (a) water concentration in a bi-phasic solvent mixture and (b) temperature were investigated to determine the optimum conditions. All reactions were carried out using free lipases and the physically adsorbed lipases in order to compare the performance with chemically conjugated nano-biomaterials. It was observed from the bio-catalytic reaction (i) that the conversion values given by lipase-immobilised materials were comparable to those given by free lipases with the added advantage of being re-usable for further catalytic cycles. PFL-immobilised nanoparticles were shown to be more effective catalysts than CRL-immobilised materials. In the bio- catalytic reaction (ii), Lipase-immobilised materials were shown to exhibit reasonable conversion values (maximum 53%) along with easy separability by one-step magnetic separation from the reaction mixture and re-usability. Finally, in the bio-catalytic reaction (iii), lipase-immobilised materials were shown to give lower total conversion values compared to free enzymes, but a higher proportion of desired products [(1S,4R)-cis-4-acetoxy-2-cyclopenten-1-ol and (1R,4S)-cis-4-acetoxy-2-cyclopenten-1-ol]. PFL (both free and immobilised) materials were shown to give higher conversion and enantioselectivity towards the desired (1S,4R)-enantiomer (93-100% ee) than CRL materials (30-40% ee). The second application involved bio-separation and bio-sensing where 5ʹ-NH2-modified oligonucleotide sequences specific to either Listeria Monocytogenes (LM) or Escherichia Coli (EC) were immobilised onto the surface of glutaraldehyde modified nanoparticles to assess the specific capture and enhance the sensitivity of detection of pathogenic bacterial DNAs from food samples. Firstly, the oligonucleotide-grafted nanoparticles were used in a hybrid capture assay (model assay) at UCLan using specific single stranded DNA primers of our interest followed by the application in real food samples at Q-Bioanalytic GmbH, Germany. Capture of the complementary sequences was reasonably high (48-70% for LM-specific materials and 48-55% for EC-specific materials) when calculated as a molar ratio of conjugated oligonucleotides to complementary oligonucleotides captured. Specific capture was determined to be 33-52% for LM-specific oligonucleotide-grafted nano-materials and 59-60% for EC-specific oligonucleotide-grafted nano-materials. Dehybridisation of captured sequences was shown to be efficient for all oligonucleotide-grafted materials (72-97% for LM-specific materials and 86-87% for EC-specific materials), indicating that the materials were ready for real applications using food matrices at Q-Bioanalytic GmbH, Germany. Nucleic acid DNA was extracted from a real food sample inoculated with either LM or EC and the extracted DNA was used for specific capture using the oligonucleotide-grafted materials tested at UCLan. Dehybridised oligonucleotides were amplified and analysed using quantitative real-time PCR (qPCR). The results showed that using a one-step hybrid capture assay, LM-specific oligonucleotide-grafted materials were successful at detecting LM from an undiluted solution of LM only and from a 1:1 mixture of LM and EC. Using a two-step assay where the forward and reverse oligonucleotide-grafted materials were applied for capture separately, only EC-specific materials were successful for the detection of EC from an undiluted solution, and also from a 1:1 mixture of LM and EC.

The aim of this research is to provide a more holistic approach to study Chumash rock art throughout their entire geographic region within South-Central California by applying geographic information systems (GIS), incorporating ethnohistoric and ethnographic data and utilising associated archaeological material under an Actor-Network Theory (ANT) framework. Through a review of past Chumash archaeological and rock art studies, I discuss where previous research is lacking and how that research was fragmentary due to focusing only on specific geographic areas or linguistic regions. As rock art is an artefact fixed within the terrain, I further argue it has a potential connection to the topography--particularly its relationship to Chumash landscapes and taskscapes by applying both formal and informed methodologies at multiple scales. By modifying the tenets of ANT to create a framework that uses the rock art data to define space, analyse its heterogeneity and connectivity and study its topographic entrenchment, this research conceptualises rock art’s networks. To conduct this research, I collated a large body of spatial and descriptive information for 254 rock art sites and associated archaeology. Spatial analyses were performed at multiple scales using GIS as a heuristic to conceptualise site clustering, landscape entrenchment and anisotropic movement for the collated data. While the rock art sites were used to define the multi-scalar spaces, results show that the identity of the sites change throughout space and time where rock art itself is a network and not exclusive to one specific Chumash network. Analysis of the data shows that the topographic setting entrenches the rock art and begins to represent the dynamic assembly of its heterogeneous network relations. Movement through the landscape reflects how the sites were connected or structured within their landscapes and taskscapes. Overall it reflects rock art’s interrelationships to the networked economic, social, ideological and political organisations of the Chumash and their rich ceremonial practices. Therefore, the Chumash rock art networks were as complex, dynamic, variable and heterogeneous as Chumash society and the rock art panels themselves.

Isolation of DNA from skeletonised human remains can be problematic. In addition to DNA degradation, enhanced by high temperature and humidity, there are often potent polymerase chain reaction (PCR) inhibitors present within the samples. It is therefore important to extract the maximum amount of available DNA whilst removing any amplification inhibitors that may be present. Whilst real-time PCR methods are available for quantification and detection of PCR inhibitors the information received is limited as real-time PCR targets amplicons that are much smaller than those typically targeted in forensic analysis. To gain more information on the quality of extracted DNA a new multiplex PCR assay comprising a 4-plex targeting amplicons of 70 base pairs (bp), 194 bp, 305 bp and 384 bp along with two Internal Amplification Contols (IACs) of 90 bp and 410 bp was developed. This multiplex was optimised so that it worked with template amounts ranging between 0.10 ng and 200 ng; partial profiles were obtained with as little as 0.02 ng. The IACs were effective in detecting PCR inhibitors. The multiplex also assessed as a quantification tool. Plotting peak height compared to input DNA of a standard dilution series produced a coefficient of determination (R2) of 0.8308. The multiplex was found to provided reasonable estimates of DNA concentration, when the sample concentration was between 12.5 – 100 ng; relative standard deviations were all below 10% in this range for 30% of tested samples. However, real-time PCR proved to be more precise and was used in the rest of the study for the purposes of quantification. In forensic cases bones and teeth often provide some of the most challenging samples to extract good quality DNA. Using the optimised multiplex to assess the quality of DNA extracts five extraction methods: ChargeSwitch® gDNA Plant Kit, DNA IQTM System Kit, DNeasy® Blood & Tissue Kit, PrepFiler® BTA Forensic DNA Extraction Kit and phenol-chloroform-isoamyl alcohol extaction methods were assessed for their capability for extracting clean DNA from bone samples. Prior to the main experimentation several evaluation studies were carried out to optimise the methods being used. Based on the results, decalcification was not used for any of the extractions as non-decalcified extracts contained higher amounts of DNA. For the phenol-chloroform-isoamyl alcohol extraction it was determined that whilst ethanol precipitation provided higher amounts of DNA, the extracts using Amicon 30kDa filters (Amicon ultra-0.5 centrifugal filter unit with ultracel-30 membrane) were cleaner. Based on poor results with degraded bone samples a pre-process technique was developed; these extractions started with 250 mg of pulverised bone sample which was then concentrated and cleaned up using Amicon 30kDa filters (Amicon ultra-2 ml centrifugal filters for DNA purification and concentration) before carrying out the standard extraction procedures. After optimisation of the extraction methods the comparison study showed that the phenol-chloroform-isoamyl alcohol extraction method produced the highest DNA yields with both fresh and degraded bone samples, followed by DNeasy® Blood & Tissue Kit, ChargeSwitch® gDNA Plant Kit, PrepFiler® BTA Forensic DNA Extraction Kit and DNA IQTM System Kit. However, all produced DNA that could be amplified and did not contain any inhibition. Another application of the multiplex was to assess the effectiveness of different DNA preservation methods by examining the amount and quality of DNA recovered after preservation. Five methods: cell lysis solution (with 1% sodium azide), dehydration / freeze drying, ethanol (96%), freezing and room temperature storage were used to study the effectiveness of preservation methods on fresh and three-month old decomposed pig bone samples which were preserved for 6 weeks, 6 months and 1 year. The results showed that freezing is the best preservation method for both fresh and degraded bone samples for long-term storage followed by ethanol (96%), dehydration / freeze drying and room temperature storage. However, full profiles were obtained from both fresh and degraded bone samples from all methods, except cell lysis solution (with 1% sodium azide). Cell lysis solution (with 1% sodium azide) preservation method tended to be good for short-term storage but with the long-term preservation, less DNA yield was obtained and also the electropherograms showed higher levels of DNA degradation. Finally, using the optimised DNA extraction methods, the multiplex was tested using forensic samples comprising of 30 bone samples from casework in Malaysia and simulated body fluid evidences subjected to environmental insult in the United Arab Emirates. The application illustrated the effectiveness of the multiplex to identify PCR inhibitors and identify DNA degradation, providing supplementary information to real-time PCR.

The decommissioning of nuclear power plants around the world will produce a major waste stream of irradiated graphite. Graphite has been used extensively as a reactor moderator and reflector material that becomes irradiated and contaminated over time. In the coming years ~250,000 tonnes of irradiated graphite will require management making this a significant waste management issue worldwide. Irradiated graphite is categorised as Intermediate Level Waste mostly due to its content of Carbon-14 (C-14) which is a long-lived radioisotope which could be released into the biosphere. In addition the Low Level Waste (LLW) repository at Drigg has very strict guidelines regarding C-14 authorisation and there is currently no deep geological repository available in the UK. Varying amounts of carbonaceous deposits have been identified on irradiated graphite samples removed from reactor cores. If these deposits are rich in C-14, treatment of the waste graphite by oxidation could reduce the C-14 inventory of the remaining graphite. This is the primary focus of this research. In order to investigate a technique that would decontaminate graphite from the carbonaceous deposits it was necessary to produce a range of carbonaceous deposits on virgin graphite material to act as a simulant for the deposits present on reactor graphite. Two deposition techniques were investigated: microwave plasma assisted chemical vapour deposition and a combination of solution deposition and charring. C-13 precursors were used as they facilitate the study of the selective removal of the deposit by mass spectrometry and spectroscopy. Using C-13 analogues instead of C-14 prevents the need to work in active laboratories and allows higher concentrations of deposit to be used which is beneficial when developing a technique for selective removal. A thermal treatment which utilised the application of a vacuum was investigated to determine whether the carbonaceous deposits could be selectively removed with minimal oxidation to the underlying graphite. As carbon deposits were more amorphous than crystalline graphite it was thought that they would oxidise quicker at lower temperatures than graphite. Virgin graphite and samples with deposits were characterised before and after thermal treatment using Scanning Electron Microscopy, Raman Spectroscopy, Thermal Gravimetric Analysis and Mass Spectrometry. An additional area of investigation was conducted using thermogravimetric studies of the oxidation of irradiated graphite which was carried out at the National Nuclear Laboratory’s Preston Lab. This would determine the distribution of C-14 in the carbonaceous deposits and underlying irradiated graphite which could be a key factor in the determination of possible treatments and eventual storage/disposal routes of the waste graphite.

A polymerase chain reaction (PCR) multiplex assay capable of amplifying 13 rapidly mutating Y chromosome short tandem repeats (RM Y-STRs) simultaneously was developed and optimised. This multiplex assay which was termed RM-Yplex is the first to include all 13 RM Y-STRs including DYF387S1, DYF399S1, DYF403S1a/b, DYF404S1, DYS449, DYS518, DYS526a/b DYS547, DYS570, DYS576, DYS612, DYS626 and DYS627. A developmental validation was performed following the Scientific Working Group for DNA Analysis Methods (SWGDAM) revised guidelines. Robustness and limitations of the assay were demonstrated through a range of studies including reproducibility, sensitivity, specificity, stability and mixture studies. Appropriate controls were used during the studies that included a number of male and female commercial controls including, 2800M, 9948 and Taqman male controls and 9947A female control. An allelic ladder was developed for the assignment of the alleles. This was done by choosing samples with different alleles, amplifying them and then adjusting the volumes of amplified products in a mixture. The developed mixtures were used to balance the composite ladder. Multiple alleles of the various loci included in the ladder were sequenced. Reference haplotypes were developed for the 5 male samples included in the Y chromosome Standard Reference Material 2395 (SRM2395) using RM-Yplex. The International Society of Forensic Genetics (ISFG) recommendations were followed for adopting allele nomenclature. As part of developmental validation, the assay was included in an external proficiency trial which was concluded successfully. An internal validation of RM-Yplex was carried out at the Department of Forensic Sciences and Criminology Laboratory, Dubai where apart from other studies; application of the assay was demonstrated using non-probative forensic casework samples. The value of RM-Yplex was demonstrated for differentiating close male relatives in a case where a previously used Y-STR multiplex assay had shown identical haplotypes for those individuals. 1160 male individual samples were analysed in this study including UAE, other Arabian Peninsula populations as well as two South Asian populations residing in United Arab Emirates. RM-Yplex haplotypes have extremely high power of discrimination. The haplotype diversity for RM-Yplex haplotype is much more than the existing commercial Y-STR assays. Population studies have been carried out for the Arab, Indian and Pakistani populations. AMOVA was conducted for determining the apportionment of diversity and pairwise FST’s were estimated between populations. These have shown a marked homogeneity within the UAE Arab sub-populations. MDS plots of pairwise FST’s indicated that populations were not grouped significantly in accordance with the geographical locations. A network analysis showed the extent of distribution of haplotypes of various populations and their relationships. A highly sensitive and reliable RM-Yplex multiplex assay has been thus developed, which is expected to help genetic populations studies and forensic casework.

The one-dimensional pyrolysis computational tool ThermaKin was used to predict the thermal decomposition behaviour of widely used synthetic polymers (polypropylene (PP) and polyethylene (PE)) with and without additives, in order to investigate the suitability of ThermaKin for novel fire retarded samples, under different thermal and fire conditions. The thermal decomposition of materials was investigated using simultaneous thermal analysis technique (STA) coupled with Fourier Transform Infrared Spectrometry (FTIR) at different heating rates and atmospheres. The results show that thermal decomposition of PP follows single mass-loss step, without formation of residue in nitrogen. It was also found that the pyrolysis shifted towards higher temperature with increase of heating rate at different atmospheres. ThermaKin fitted the TGA curves very well. The thermal decomposition behaviour of polypropylene grafted with 5wt% of maleic anhydride (MA), and reinforced with 5wt% of closite 20A as nanoclay (PP-gMA/NC) was also investigated. The main conclusions from this data are that during the thermal decomposition in different atmospheres, TGA curves showed a single step of decomposition process for all samples. The effect of clay is more pronounced during thermal oxidation. In N2 and air, a two-step reaction mechanism was fitted the experimental curves fairly well. The thermal decomposition of PE, pure and reinforced with different types of carbon fillers (single/multi wall carbon nanotubes, carbon fibres, carbon black and single/few layers of graphene nanosheets), at different loadings (0.1, 0.5 and 1 wt%) and atmospheres were investigated, to determine their suitability as potential fire retardant additives. Results showed that thermal decomposition of PE and its composites/nanocomposites followed a single mass-loss step at a range of temperatures, with no residue formation in N2. The DTG curve in air showed two mass loss rate peaks. The experimental results showed that all loadings of these different additives made no improvement to the thermal stability of PE/MA. In air, the compatibilising agent (MA) improved the thermal stability of pure PE, compared to these composites/nanocomposites at the selected loadings. Mechanisms of single or two-step reaction in N2, and three-step reaction in air for the thermal decomposition of PE with and without additives predicted fairly well the experimental curves. Finally, the work was extended to investigate the performance of ThermaKin to establish a model that is able to predict cone calorimetry results. ThermaKin predicted the burning rate of PE/MA, as a good agreement between the experimental and simulated curves was achieved. Sensitivity analysis was performed to investigate the influence of the variation of the material properties on the modelling results. It was found that the heat of decomposition is the most important parameter of those investigated and needs to be determined most accurately. Heat capacity and thermal conductivity are somewhat important. The absorption coefficient and the reflectivity are of lesser importance. In conclusion, this work shows that the combination of pyrolysis modelling, thermal and chemical analysis techniques provides a strong and powerful tool for generating a comprehensive understanding of the thermal decomposition of novel fire retardant materials. However, further work is needed to study the influence of the changes of the material properties in polymeric material while reinforced with different additives and how this will be reflected on the modelling parameters and mechanism.

Studying the spermatogenesis of horse is beneficial for the horse industry by identifying the causes of chromosomal abnormalities, which cause embryonic loss, congenital abnormalities and infertility. Little is known about the spermatogenesis in horse. This is the first report that investigates the horse spermatogenesis in detail, particularly metaphase I (MI) and prophase I (PI) stages of the first meiotic division. Meiotic recombination is considered to be the major outcome of meiosis. It is essential for proper chromosome segregation and formation of normal haploid gametes. Analysis of recombination frequency and distribution are crucial for genomic and association studies. Any alteration of the recombination frequency and positioning can cause non-disjunction and generation of aneuploidy. The frequency and distribution of chiasmata were estimated at MI chromosomes from fourteen fertile stallions. The average frequency of autosomal chiasmata was 49.45 ± 2.07, corresponding to a genetic length of 2,472.5 cM. All autosomal bivalents had at least one chiasma. The majority of chromosomes have one or two chiasmata, which are mostly distally localized. The frequency and the distribution as well as the genetic length of chiasmata were also estimated for the first time in eight different individual autosomes. Immunofluorescent localization was used to characterize the early stages of the first meiotic division as well as to examine the frequency and the distribution of DNA mismatch repair protein MutL Homologous Protein 1 (MLH1) foci on synaptonemal complexes (SCs) from sex fertile stallions. The mean frequency of autosomal recombination foci was 50.11±2.35. All autosomal bivalents had at least one recombination focus. In general, foci were located near the distal ends with some foci interstitially distributed. The distribution of MLH1 foci indicated positive interference; however, foci were very close to one another in rare instances. The average SCs relative length was highly correlated with the average number of MLH1 foci. MLH1 have been proposed to mark crossover sites at PI since the frequency and distribuation of MLH1 foci closely correspond to the frequency and distribution of chiasmata on MI chromosomes. iii | P a g e Spermatozoa viability, which include spermatozoa head and tail membrane integrity, acrosomal integrity and mitochondrial function assessment are the main sperm analysis parameters considered in this thesis to evaluate the stallion fertility using epididymal collected semen samples. The mean percentage of spermatozoa with viable heads and tails, using Chicago sky blue stain, was 81.26 ± 5.06. FITC-Pisum sativum agglutinin (FITC-PSA) and MitoTracer green were used successfully to assess the spermatozoal acrosomal status as well as the mitochondrial function, respectively. The mean percentage of spermatozoa with integrated acrosome was 93.85 ± 1.9, while for functional mitochondria was 95.63 ± 1.63. In conclusion, this finding is the cornerstone to understanding the genetic basis of normal horse spermatogenesis. Simultaneous assessment of different functional sperm parameters as well as investigating the synapsis and recombination frequency and distribution, at PI or MI, would assist with predictions of stallion fertility prior to breeding. In addition, this study will enable investigators to use linkage analysis in identifying and localising different genetic loci associated with specific traits.

Forensic science is growing rapidly in the world today. During the past decade, medico-legal investigations have been highly expanded to include all areas of forensic science. The present study investigated three important aspects of forensic biology. First, this present project investigated, a total number of 5,703 reported medico-legal cases diagnosed as un-natural deaths by The Forensic Department in Kuwait, during the year 2003-2009. The results show that accidental, homicidal and suicidal deaths accounted for 86%, 8% and 6%, respectively. The results showed that most people who died of unnatural deaths were more predominant in the age group 20-29 years (third decade). Road Traffic Accidents accounted for 65% of accidental deaths, and 4% out of them were related to alcohol consumption. The results also illustrated that the highest rate of homicide in Kuwait was due to stab wound injuries (38%) compared to the lower rate of homicidal pattern for infanticides (3%). Similarly, the study showed that the most common method of suicide in Kuwait was death by hanging and this accounted for (60%). This study further demonstrated the effectiveness of virtual autopsy technique as a new tool in forensic investigations to determine various un-natural death causes. A total of thirty (30) male forensic cadavers were employed in this project. The cases were RTA (11), firearm injuries (10), drowning (4), head injuries (3) and lastly strangulation (2). All these cases were compared to the findings of traditional autopsy. The results show similar findings for virtopsy compared to traditional autopsy. This study clearly revealed that virtopsy could be an effective alternative in certain situation, being noninvasive and rapid. The present project also investigated 28 samples of human blood, saliva or semen. The experiments were done at four different temperatures (55°C, 37°C, 24°C and 4°C) and four different humidity ranges (41%, 55%, 58% and 61%), respectively. The results showed that, DNA quantity in blood, saliva and semen samples remained more or less the same at temperatures of 4°C, 24°C and 37°C compared to values for day one with all other days. In contrast, when the temperature was raised to 55 °C, the DNA started to degrade with time until it reached zero at day 12 for saliva and day 15 for blood, but not for semen. The results clearly show that DNA in saliva and blood samples is extremely sensitive to heat compared to semen. In conclusion, the study reveals the different causes of unnatural deaths, the value of virtual autopsy and the need for early DNA measurement in Kuwait.

The burden of Diabetes Mellitus (DM) is increasing worldwide and it is estimated to reach indefinite proportions of about 450 million by year 2030. Patients with type 2 diabetes mellitus (T2DM) have a significantly increased risk of developing cardiovascular diseases (CVD). Moreover, CVD is the major cause of mortality and morbidity (75%) in T2DM patients. DM itself has been long recognised as an independent risk factor for several forms of CVD including coronary heart disease (CHD), peripheral arterial disease, cardiomyopathy and congestive heart failure in both men and women. It is well-known that T2DM is associated with several factors including hyperglycaemia, hypertension, dyslipidemia, obesity all of which contribute to CVD. In order to prevent CVD, early intervention on cardiovascular risk factors is vital during clinical assessment of T2DM patients. A major role of inflammation has been well described in the development of CVD in T2DM patients. Inflammatory process and factors which contribute to CVD in T2DM patients have recently become a focus in diabetic research. Elucidation of common patho-physiological mechanisms among T2DM patients might emphasize the role of inflammation in CVD. The main purpose of this study was to investigate any patho-physiological changes in red blood cells (RBC), white blood cells (neutrophils and lymphocytes) and plasma, measuring RBC membrane fragility and proteins, intracellular free calcium concentrations [Ca2+]i and several cations including Na+, Mg2+, Ca2+, Fe2+, Zn2+ and Cu2+, biochemical parameters and inflammatory mediators which normally serve as independent predisposing risk factors for CVD among T2DM patients compared to age-match healthy controls. The results have shown that fura-2 loaded neutrophils and lymphocytes in blood from T2DM patients contain significantly (p<0.05) less [Ca2+]i than neutrophils and lymphocytes from healthy subjects upon stimulation with physiological doses of either fMLP or thapsigargin indicating a derangement in cellular calcium homeostasis during T2DM. Similarly, RBC membranes from T2DM patients contained significantly (p<0.05) more spectrin, ankyrin, band 3, band 4.1, glycophorin etc compared to RBC membranes from age-matched healthy control subjects. The results also show that the RBCs from T2DM patients were more fragile compared to RBC from healthy controls. Measurement of protein glycation in plasma have revealed significantly (p<0.05) more fluorescence in proteins form T2DM patients compared to control. In relation to plasma cations and intracellular markers and mediators, the results show that plasma from T2DM patients contain significantly (p<0.05) more Na+, Mg2+ , Ca2+, Fe2+, Zn2+ and Cu2+ compared to plasma levels from age-match healthy controls. Similarly, the concentrations of kidney and liver function markers such as urea, creatinine, alkaline phosphatase, ALT, AST, GGT, total protein and albumin increased significantly (p<0.05) compared to healthy controls. The same is also true for glucose, total cholesterol, triglycerides, CRP, HBA1C, WBC where the blood from T2DM patients contained elevated concentrations compared to blood from healthy age-matched control patients. Together, the results of this study have clearly demonstrated marked and significant changes in cellular calcium homeostasis in white blood cells, RBC membrane proteins and fragility, plasma protein glycation and in plasma levels of cations, intracellular markers and mediators of T2DM patients compared to healthy controls. Therefore, it is proposed that an early integrated and multi-factorial intervention of risk factors and inflammatory markers must be done in order to reduce the risk of CVD and possible mortality of T2DM patients.