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The dentate gyrus has often been posited to act as a gate that dampens highly active afferent input into the hippocampus. Effective gating is thought to prevent seizure initiation and propagation in the hippocampus and support learning and memory processes. Pathological changes to DG circuitry that occur in temporal lobe epilepsy (TLE) can increase DG excitability and impair its gating ability which can contribute to seizures and cognitive deficits. There is evidence that TLE pathologies and seizures may independently contribute to learning and memory deficits in TLE through distinct mechanisms. These two factors are difficult to untangle since TLE pathologies can drive seizures, and seizures can worsen TLE pathologies. Here we assessed whether chemogenetically increasing dentate granule cell (DGC) excitability was enough to break down the dentate gate in the absence of TLE pathologies. We found that increasing excitability specifically in DGCs caused seizures in non-epileptic mice. Importantly, due to the modulatory nature of DREADD effects, seizures were driven by intrinsic circuit activity rather than direct activation of DGCs. These seizures resulted in a spatial memory deficit when induced after training in the spatial object recognition task and showed stereotypical patterns of activity in miniscope calcium recordings. Our results provide direct support for the dentate gate hypothesis since seizures could be induced in non-epileptic animals by artificially degrading the dentate gate with chemogenetics in the absence of epilepsy pathologies.

Traumatic brain injury (TBI) often results in persistent learning and memory deficits, likely due to disrupted hippocampal circuitry underlying these processes. Precise temporal control of hippocampal neuronal activity is important for memory encoding and retrieval and is supported by oscillations that dynamically organize single unit firing. Using high-density laminar electrophysiology, we discovered a loss of oscillatory power across CA1 lamina, with a profound, layer-specific reduction in theta-gamma phase amplitude coupling in injured rats. Interneurons from injured animals were less strongly entrained to theta and gamma oscillations, suggesting a mechanism for the loss of coupling, while pyramidal cells were entrained to a later phase of theta. During quiet immobility, we report decreased ripple amplitudes from injured animals during sharp-wave ripple events. These results reveal deficits in information encoding and retrieval schemes essential to cognition that likely underlie TBI-associated learning and memory impairments, and elucidate potential targets for future neuromodulation therapies.

Cognitive impairment is a common symptom following mild traumatic brain injury (mTBI or concussion) and can persist for years in some individuals. Hippocampal slice preparations following closed-head, rotational acceleration injury in swine have previously demonstrated reduced axonal function and hippocampal circuitry disruption. However, electrophysiological changes in hippocampal neurons and their subtypes in a large animal mTBI model have not been examined. Using in vivo electrophysiology techniques, we examined laminar oscillatory field potentials and single unit activity in the hippocampal network 7 days post-injury in anesthetized minipigs. Concussion altered the electrophysiological properties of pyramidal cells and interneurons differently in area CA1. While the firing rate, spike width and amplitude of CA1 interneurons were significantly decreased post-mTBI, these parameters were unchanged in CA1 pyramidal neurons. In addition, CA1 pyramidal neurons in TBI animals were less entrained to hippocampal gamma (40-80 Hz) oscillations. Stimulation of the Schaffer collaterals also revealed hyperexcitability across the CA1 lamina post-mTBI. Computational simulations suggest that reported changes in interneuronal physiology may be due to alterations in voltage-gated sodium channels. These data demonstrate that a single concussion can lead to significant neuronal and circuit level changes in the hippocampus, which may contribute to cognitive dysfunction following mTBI., (© 2023. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)

Brain organoids created from human pluripotent stem cells represent a promising approach for brain repair. They acquire many structural features of the brain and raise the possibility of patient-matched repair. Whether these entities can integrate with host brain networks in the context of the injured adult mammalian brain is not well established. Here, we provide structural and functional evidence that human brain organoids successfully integrate with the adult rat visual system after transplantation into large injury cavities in the visual cortex. Virus-based trans-synaptic tracing reveals a polysynaptic pathway between organoid neurons and the host retina and reciprocal connectivity between the graft and other regions of the visual system. Visual stimulation of host animals elicits responses in organoid neurons, including orientation selectivity. These results demonstrate the ability of human brain organoids to adopt sophisticated function after insertion into large injury cavities, suggesting a translational strategy to restore function after cortical damage., Competing Interests: Declaration of interests G.-l.M. is on the editorial board of Cell Stem Cell., (Published by Elsevier Inc.)

Carbonaceous asteroids, such as (101955) Bennu, preserve material from the early Solar System, including volatile compounds and organic molecules. We report spacecraft imaging and spectral data collected during and after retrieval of a sample from Bennu's surface. The sampling event mobilized rocks and dust into a debris plume, excavating a 9-meter-long elliptical crater. This exposed material is darker, spectrally redder, and more abundant in fine particulates than the original surface. The bulk density of the displaced subsurface material was 500 to 700 kilograms per cubic meter, which is about half that of the whole asteroid. Particulates that landed on instrument optics spectrally resemble aqueously altered carbonaceous meteorites. The spacecraft stored 250 ± 101 grams of material, which will be delivered to Earth in 2023.

We investigate the shape of near-Earth asteroid (101955) Bennu by constructing a high-resolution (20 cm) global digital terrain model from laser altimeter data. By modeling the northern and southern hemispheres separately, we find that longitudinal ridges previously identified in the north extend into the south but are obscured there by surface material. In the south, more numerous large boulders effectively retain surface materials and imply a higher average strength at depth to support them. The north has fewer large boulders and more evidence of boulder dynamics (toppling and downslope movement) and surface flow. These factors result in Bennu's southern hemisphere being rounder and smoother, whereas its northern hemisphere has higher slopes and a less regular shape. We infer an originally asymmetric distribution of large boulders followed by a partial disruption, leading to wedge formation in Bennu's history., (Copyright © 2020 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution License 4.0 (CC BY).)

While hippocampal-dependent learning and memory are particularly vulnerable to traumatic brain injury (TBI), the functional status of individual hippocampal neurons and their interactions with oscillations are unknown following injury. Using the most common rodent TBI model and laminar recordings in CA1, we found a significant reduction in oscillatory input into the radiatum layer of CA1 after TBI. Surprisingly, CA1 neurons maintained normal firing rates despite attenuated input, but did not maintain appropriate synchronization with this oscillatory input or with local high-frequency oscillations. Normal synchronization between these coordinating oscillations was also impaired. Simultaneous recordings of medial septal neurons known to participate in theta oscillations revealed increased GABAergic/glutamatergic firing rates postinjury under anesthesia, potentially because of a loss of modulating feedback from the hippocampus. These results suggest that TBI leads to a profound disruption of connectivity and oscillatory interactions, potentially disrupting the timing of CA1 neuronal ensembles that underlie aspects of learning and memory., (Copyright © 2020 Koch et al.)

The OSIRIS-REx mission has observed multiple instances of particles being ejected from the surface of near-Earth asteroid (101955) Bennu. The ability to quickly identify the particle trajectories and origins is necessary following a particle ejection event. Using proven initial orbit determination techniques, we can rapidly estimate particle trajectories and ejection locations. We present current results pertaining to the identification of particle tracks, an evaluation of the estimated orbits and the excess velocity necessary to induce the particle ejection from the surface, and the uncertainty quantification of the ejection location. We estimate energies per particle ranging from 0.03 to 11.03 mJ for the largest analyzed events and velocities ranging from 5 to 90 cm/s, though we exclude the highest-velocity particles in this technique. We estimate ejection times for eight events and constrain six of the analyzed ejection events to have occurred between about 16:30 and 19:00 local solar time, with the largest events occurring between 16:30 and 18:05., (©2020. The Authors.)

Near-Earth asteroid (101955) Bennu is an active asteroid experiencing mass loss. The activity manifests itself in the form of ejection events emitting up to hundreds of millimeter- to centimeter-scale particles. The Origins, Spectral Interpretation, Resource Identification, and Security-Regolith Explorer spacecraft monitored particle activity for a 10-month period that included Bennu's perihelion and aphelion. Novel and classical methods were utilized to detect the particles and characterize their orbital and physical properties. Roughly 30% of the observed particle mass escaped to heliocentric orbit. A majority of particles fell back onto the surface of Bennu after ejection, with the longest-lived particle surviving for 6 days on a temporary orbit. Particle ejection events appear to preferentially take place in the afternoon and evening and from low latitudes, although they can occur at any time or latitude. The reaccumulation of material is biased toward low latitudes resulting in the possible in-fill of craters and growth of Bennu's equatorial bulge. Of the potential mechanisms behind this activity that were investigated in focused studies, meteoroid impacts, thermal fracturing, and ricochet-but not water ice sublimation-were found to be consistent with observations. While phyllosilicate dehydration was not investigated with a focused study, it remains a possible mechanism. These mechanisms are not unique to Bennu, suggesting that many near-Earth asteroids may exhibit activity that has gone undetected thus far. Spacecraft missions with wide-field imagers are encouraged to further characterize this phenomenon., (©2020. The Authors.)

OSIRIS-REx began observing particle ejection events shortly after entering orbit around near-Earth asteroid (101955) Bennu in January 2019. For some of these events, the only observations of the ejected particles come from the first two images taken immediately after the event by OSIRIS-REx's NavCam 1 imager. Without three or more observations of each particle, traditional orbit determination is not possible. However, by assuming that the particles all ejected at the same time and location for a given event, and approximating that their velocities remained constant after ejection (a reasonable approximation for fast-moving particles, i.e., with velocities on the order of 10 cm/s or greater, given Bennu's weak gravity), we show that it is possible to estimate the particles' states from only two observations each. We applied this newly developed technique to reconstruct the particle ejection events observed by the OSIRIS-REx spacecraft during orbit about Bennu. Particles were estimated to have ejected with inertial velocities ranging from 7 cm/s to 3.3 m/s, leading to a variety of trajectory types. Most (>80%) of the analyzed events were estimated to have originated from midlatitude regions and to have occurred after noon (local solar time), between 12:44 and 18:52. Comparison with higher-fidelity orbit determination solutions for the events with sufficient observations demonstrates the validity of our approach and also sheds light on its biases. Our technique offers the capacity to meaningfully constrain the properties of particle ejection events from limited data., (©2020. The Authors.)

Human brain organoids provide unique platforms for modeling development and diseases by recapitulating the architecture of the embryonic brain. However, current organoid methods are limited by interior hypoxia and cell death due to insufficient surface diffusion, preventing generation of architecture resembling late developmental stages. Here, we report the sliced neocortical organoid (SNO) system, which bypasses the diffusion limit to prevent cell death over long-term cultures. This method leads to sustained neurogenesis and formation of an expanded cortical plate that establishes distinct upper and deep cortical layers for neurons and astrocytes, resembling the third trimester embryonic human neocortex. Using the SNO system, we further identify a critical role of WNT/β-catenin signaling in regulating human cortical neuron subtype fate specification, which is disrupted by a psychiatric-disorder-associated genetic mutation in patient induced pluripotent stem cell (iPSC)-derived SNOs. These results demonstrate the utility of SNOs for investigating previously inaccessible human-specific, late-stage cortical development and disease-relevant mechanisms., Competing Interests: Declaration of Interests The authors declare no competing interests., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Active asteroids are those that show evidence of ongoing mass loss. We report repeated instances of particle ejection from the surface of (101955) Bennu, demonstrating that it is an active asteroid. The ejection events were imaged by the OSIRIS-REx (Origins, Spectral Interpretation, Resource Identification, and Security-Regolith Explorer) spacecraft. For the three largest observed events, we estimated the ejected particle velocities and sizes, event times, source regions, and energies. We also determined the trajectories and photometric properties of several gravitationally bound particles that orbited temporarily in the Bennu environment. We consider multiple hypotheses for the mechanisms that lead to particle ejection for the largest events, including rotational disruption, electrostatic lofting, ice sublimation, phyllosilicate dehydration, meteoroid impacts, thermal stress fracturing, and secondary impacts., (Copyright © 2019 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.)

The formation and release under gravity of blood droplets from simulated weapon tips has been investigated experimentally and the results analysed and interpreted using established theoretical models for detached pendent drops. Droplets were produced from a series of conical nozzles, manufactured with a range of cone angles and including a set of un-bored conical tips, under conditions where the tip was either non-wetted or pre-wetted with blood. For the former, radius-limited case, detached droplet volumes were found to agree well with the predictions of both the pendent drop weight and drop shape models. For pre-wetted tips, droplet volumes were found to increase with increasing cone angle and to be independent of whether the blood flow was through an orifice at the tip or across the tip surface itself for un-bored tips. Such angle-limited, detached droplet volumes were predicted well by applying the same contact angle correction factor as for a flat surface. The maximum droplet volume, formed from a horizontal flat surface, was found to be around 150μL for horse blood. Similar theoretical approaches were then extended to droplets released under centrifugal force appropriate to swing cast-off activity and evaluated using previously published experimental data. Order of magnitude agreement for droplet diameters was fairly good but, more importantly, these were found to be proportional to the inverse of the tangential velocity thereby supporting a model where the blood droplet is released directly from a blood mass itself, such as a ligament, rather than from being pinned to some surface feature or orifice on the weapon. This work provides a sound theoretical and experimental understanding of blood droplet release under these conditions that can underpin both future research and the interpretation of blood evidence in case-work., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Decoding laminar information across deep brain structures and cortical regions is necessary in order to understand the neuronal ensembles that represent cognition and memory. Large animal models are essential for translational research due to their gyrencephalic neuroanatomy and significant white matter composition. A lack of long-length probes with appropriate stiffness allowing penetration to deeper structures with minimal damage to the neural interface is one of the major technical limitations to applying the approaches currently utilized in lower order animals to large animals. We therefore tested the performance of multichannel silicon probes of various solutions and designs that were developed specifically for large animal electrophysiology. Neurophysiological signals from dorsal hippocampus were recorded in chronically implanted awake behaving Yucatan pigs. Single units and local field potentials were analyzed to evaluate performance of given silicon probes over time. EDGE-style probes had the highest yields during intra-hippocampal recordings in pigs, making them the most suitable for chronic implantations and awake behavioral experimentation. In addition, the cross-sectional area of silicon probes was found to be a crucial determinant of silicon probe performance over time, potentially due to reduction of damage to the neural interface. Novel 64-channel EDGE-style probes tested acutely produced an optimal single unit separation and a denser sampling of the laminar structure, identifying these research silicon probes as potential candidates for chronic implantations. This study provides an analysis of multichannel silicon probes designed for large animal electrophysiology of deep laminar brain structures, and suggests that current designs are reaching the physical thresholds necessary for long-term (∼1 month) recordings with single-unit resolution.

The shapes of asteroids reflect interplay between their interior properties and the processes responsible for their formation and evolution as they journey through the Solar System. Prior to the OSIRIS-REx (Origins, Spectral Interpretation, Resource Identification, and Security-Regolith Explorer) mission, Earth-based radar imaging gave an overview of (101955) Bennu's shape. Here, we construct a high-resolution shape model from OSIRIS-REx images. We find that Bennu's top-like shape, considerable macroporosity, and prominent surface boulders suggest that it is a rubble pile. High-standing, north-south ridges that extend from pole to pole, many long grooves, and surface mass wasting indicate some low levels of internal friction and/or cohesion. Our shape model indicates that, similar to other top-shaped asteroids, Bennu formed by reaccumulation and underwent past periods of fast spin leading to its current shape. Today, Bennu might follow a different evolutionary pathway, with interior stiffness permitting surface cracking and mass wasting.

The hippocampus is integral to working and episodic memory and is a central region of interest in diseases affecting these processes. Pig models are widely used in translational research and may provide an excellent bridge between rodents and nonhuman primates for CNS disease models because of their gyrencephalic neuroanatomy and significant white matter composition. However, the laminar structure of the pig hippocampus has not been well characterized. Therefore, we histologically characterized the dorsal hippocampus of Yucatan miniature pigs and quantified the cytoarchitecture of the hippocampal layers. We then utilized stereotaxis combined with single-unit electrophysiological mapping to precisely place multichannel laminar silicon probes into the dorsal hippocampus without the need for image guidance. We used in vivo electrophysiological recordings of simultaneous laminar field potentials and single-unit activity in multiple layers of the dorsal hippocampus to physiologically identify and quantify these layers under anesthesia. Consistent with previous reports, we found the porcine hippocampus to have the expected archicortical laminar structure, with some anatomical and histological features comparable to the rodent and others to the primate hippocampus. Importantly, we found these distinct features to be reflected in the laminar electrophysiology. This characterization, as well as our electrophysiology-based methodology targeting the porcine hippocampal lamina combined with high-channel-count silicon probes, will allow for analysis of spike-field interactions during normal and disease states in both anesthetized and future awake behaving neurophysiology in this large animal.

Metabonomic profiling techniques, with established quality control methods, have been used to detect subtle metabolic differences in tissue that could aid in the discovery of fraud within the food industry. Liquid chromatography quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS) was utilized to acquire metabolic profiles of muscle, heart, and liver tissue from normally slaughtered and dead on arrival chickens. A workflow including XCMS Online for data processing and robust confirmatory statistics was used in order to differentiate between the two sample types. It was found that normally slaughtered and dead on arrival chicken can be differentiated based on the metabolic profile and multivariate analysis. Markers were found to be significantly different between the two sample types in all samples. With the use of the METLIN database and MS/MS analysis of chemical standards, sphingosine was identified as a marker in the muscle tissue samples which may offer potential for the detection of fraudulently processed chicken meat. The approach taken in this work has shown that it is possible to apply the described workflows to food fraud problems, with an objective of identifying key markers that could be investigated further to determine their usefulness for fraud detection.

Cuticular hydrocarbons were extracted daily from the larvae of two closely related blowflies Calliphora vicina and Calliphora vomitoria (Diptera: Calliphoridae). The hydrocarbons were then analysed using Gas Chromatography-Mass Spectrometry (GC-MS), with the aim of observing changes within their chemical profiles in order to determine the larval age. The hydrocarbons were examined daily for each species from 1day old larvae until pupariation. The results show significant chemical changes occurring from the younger larvae to the post-feeding larvae. With the aid of a multivariate statistical method (Principal Component Analysis and Artificial Neural Networks), samples were clustered and classified, allowing for the larval age to be established. Results from this study allowed larvae to be aged to the day with at worst, 87% accuracy, which suggests there is great potential for the use of cuticular hydrocarbons present on larvae to give an indication of their age and hence potentially a valuable tool for minimum PMI estimations., (Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.)

Calliphoridae are known to be the most forensically important insects when it comes to establishing the minimum post mortem interval (PMImin) in criminal investigations. The first step in calculating the PMImin is to identify the larvae present to species level. Accurate identification which is conventionally carried out by morphological analysis is crucial because different insects have different life stage timings. Rapid identification in the immature larvae stages would drastically cut time in criminal investigations as it would eliminate the need to rear larvae to adult flies to determine the species. Cuticular hydrocarbon analysis on 1st instar larvae has been applied to three forensically important blowflies; Lucilia sericata, Calliphora vicina and Calliphora vomitoria, using gas chromatography-mass spectrometry (GC-MS) and principal component analysis (PCA). The results show that each species holds a distinct "fingerprint" hydrocarbon profile, allowing for accurate identification to be established in 1-day old larvae, when it can be challenging to apply morphological criteria. Consequently, this GC-MS based technique could accelerate and strengthen the identification process, not only for forensically important species, but also for other entomological samples which are hard to identify using morphological features., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

In analytical chemistry large datasets are collected using a variety of instruments for multiple tasks, where manual analysis can be time-consuming. Ideally, it is desirable to automate this process while obtaining an acceptable level of accuracy, two aims that artificial neural networks (ANNs) can fulfil. ANNs possess the ability to classify novel data based on their knowledge of the domain to which they have been exposed. ANNs can also analyse non-linear data, tolerate noise within data and are capable of reducing time taken to classify large amounts of novel data once trained, making them well-suited to the field of analytical chemistry where large datasets are present (such as that collected from gas chromatography-mass spectrometry (GC-MS)). In this study, the use of ANNs for the autonomous analysis of GC-MS profiles of Lucilia sericata larvae is investigated, where ANNs are required to estimate the age of the larvae to aid in the estimation of the post mortem interval (PMI). Two ANN analysis approaches are presented, where the ANN correctly classified the data with accuracy scores of 80.8% and 87.7% and Cohen's Kappa coefficients of 0.78 and 0.86. Inspection of these results shows the ANN to confuse two consecutive days which are of the same life stage and as a result are very similar in their chemical profile, which can be expected. The grouping of these two days into one class further improved results where accuracy scores 89% and 97.5% were obtained for the two analysis approaches., (Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.)

The spreading of bloodstains on matt, vinyl silk and gloss painted surfaces, following both perpendicular and non-perpendicular impact, has been studied with a view to understanding whether surface-specific properties affect the size and shape of the final stain. In all cases the dimensions of the stains on the rougher, matt paint are up to 5% less than on the other painted surfaces though this difference decreases with impact velocity. Both the diameter of circular stains and the width of elliptical stains have been successfully modelled as a function of the perpendicular component of impact velocity vn using a modification of the energy conservation approach through the inclusion of a single, empirical constant characteristic of the surface itself. By further development of this theory, the characteristic dynamical length parameter which governs the spreading has been modified to be dependent on angle thus enabling the length of elliptical stains to be predicted independently for the first time. For all these surfaces, experiment shows that the impact angles, calculated using the conventional trigonometric formula, are in deficit by around 1° at high vn increasing to around 5° at lower vn. By modelling this deficit independently using the theoretical expressions for stain width and length, this variation with vn has been successfully explained. This work has demonstrated some of the fundamental systematic sources of error in using the conventional formula for interpreting elliptical stains and established some of the basic theory on which to develop the interpretation of casework stains on surfaces, in the future., (Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.)

Previous studies on Diptera have shown the potential for the use of cuticular hydrocarbons' analysis in the determination of larval age and hence the postmortem interval (PMI) for an associated cadaver. In this work, hydrocarbon compounds, extracted daily until pupation from the cuticle of the blowfly Lucilia sericata (Diptera: Calliphoridae), have been analyzed using gas chromatography-mass spectrometry (GC-MS). The results show distinguishing features within the hydrocarbon profile over the period of the larvae life cycle, with significant chemical changes occurring from the younger larvae to the postfeeding larvae. Further interpretation of the chromatograms using principal component analysis revealed a strong correlation between the magnitudes of particular principal components and time. This outcome suggests that, under the conditions of this study, the cuticular hydrocarbons evolve in a systematic fashion with time, thus supporting the potential for GC-MS analysis as a tool for establishing PMI where such a species is present., (© 2012 American Academy of Forensic Sciences.)

A detailed understanding of blood droplet impact dynamics and stain formation is an essential prerequisite to the interpretation of both individual bloodstains and spatter patterns. The current literature on theoretical models for the spreading and splashing of liquid drops on surfaces relevant to the forensic context of bloodstain formation has been reviewed. These models have been evaluated for a paper substrate using experimental data obtained as function of droplet size, impact velocity and angle. It is shown that for perpendicular impact there are fairly simple mathematical models for the spreading diameter and the number of scallops or spines formed around the stain though these have quite limited ranges of validity in their basic form. In particular, predictions for the diameter are best for small droplets impacting at high velocity and the number of spines saturates for higher impact velocities. In the case of spreading, a modification to the energy conservation model is found to provide excellent agreement with experimental stain diameters across a wide range of impact velocities. For non-perpendicular impact, the width of stains is found to depend principally on the normal component of impact velocity and may be predicted by an appropriate modification to the expression for the perpendicular case. Limitations in the calculation of impact angle from the stain aspect ratio are identified and a theoretical basis for the prediction of spines around an elliptical stain is proposed. Some key issues for future research are identified which include a systematic, quantitative study of the effect of surface properties on bloodstain formation., (Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.)

A novel approach to the non-destructive discrimination of black, ball-point pen ink-lines on paper, has been developed which uses a standard luminescence spectrometer coupled with multivariate statistical analysis. The application of Principal Component Analysis (PCA) to these data-sets, followed by linear regression of the loadings, not only facilitates separation of the ink luminescence from that of the paper, but also allows a direct comparison between two ink lines on documents. This data reduction means that such a comparison can be done on an effectively univariate basis whereby a straightforward statistical comparison is made between two numbers, each characteristic of an ink. Thus the question, in the forensic context, of whether two ink-lines are indistinguishable may be answered with a particular statistical confidence and the method shows significantly better discrimination, for the pens studied, than conventional forensic investigative techniques involving the luminescence imaging of writing using filtered light.

The technique of principal component analysis has been applied to the UV-vis spectra of inks obtained from a wide range of black ballpoint pens available in the UK market. Both the pen ink and material extracted from the ink line on paper have been examined. Here, principal component analysis characterised each spectrum within a group through the numerical loadings attached to the first few principal components. Analysis of the spectra from multiple measurements on the same brand of pen showed excellent reproducibility and clear discrimination between inks that was supported by statistical analysis. Indeed it was possible to discriminate between the pen ink and the ink line from all brands examined in this way, suggesting that the solvent extraction process may have an influence on these results. For the complete set of 25 pens, interpretation of the loadings for the first few principal components showed that both the pen inks and the extracted ink lines may be classified in an objective manner and in agreement with the results of parallel thin layer chromatography studies. Within each class almost all inks could be individualised. Further work has shown that principal component analysis may be used to identify a particular ink from a database of reference UV-vis spectra and a strategy for developing this approach is suggested.

Thermally sprayed hydroxyapatite powders and coatings have been studied using X-ray powder diffraction and scanning electron microscopy. Preferred orientation of the crystallites has been found on a number of coated plates using X-ray powder diffraction, and full profile fitting of the patterns from the extracted powders indicates anisotropically thermally strained crystallites. This is a result of rapid crystal regrowth and associated thermal gradients on deposition. Electron micrographs of some coatings show a consistent picture of columnar crystallites orientated perpendicular to the substrate surface.