• A review of the worldwide prevalence of vitiligo in children/adolescents and adults.

      Krüger, Christian; Schallreuter, Karin U. (2012-10)
      Background  Vitiligo is an acquired, idiopathic, and worldwide common depigmentation disorder with an estimated prevalence from 0.1 to 8%. These numbers are based on clinical population studies and field research examining inhabitants of geographically enclosed areas. Our aim was to collect all available data on the prevalence of vitiligo in the general population, paying particular attention to children/adolescent groups and adults. Materials and methods  Screening of available literature and online databases using several key words. Results  We found more than 50 studies that used several methods and subgroups of the general population. The prevalence of vitiligo ranges from 0.06 to 2.28%, whereas this was 0.0–2.16% in children/adolescents populations. Conclusions  The often cited prevalence of 8% could not be confirmed after excluding clinical patient populations. Accordingly, the worldwide prevalence of vitiligo ranges between 0.5 and 2%.
    • Revisiting the Blind Tests in Crystal Structure Prediction: Accurate Energy Ranking of Molecular Crystals.

      Asmadi, Aldi; Neumann, M.A.; Kendrick, John; Girard, P.; Perrin, M-A.; Leusen, Frank J.J. (American Chemical Society, 01/12/2009)
      In the 2007 blind test of crystal structure prediction hosted by the Cambridge Crystallographic Data Centre (CCDC), a hybrid DFT/MM method correctly ranked each of the four experimental structures as having the lowest lattice energy of all the crystal structures predicted for each molecule. The work presented here further validates this hybrid method by optimizing the crystal structures (experimental and submitted) of the first three CCDC blind tests held in 1999, 2001, and 2004. Except for the crystal structures of compound IX, all structures were reminimized and ranked according to their lattice energies. The hybrid method computes the lattice energy of a crystal structure as the sum of the DFT total energy and a van der Waals (dispersion) energy correction. Considering all four blind tests, the crystal structure with the lowest lattice energy corresponds to the experimentally observed structure for 12 out of 14 molecules. Moreover, good geometrical agreement is observed between the structures determined by the hybrid method and those measured experimentally. In comparison with the correct submissions made by the blind test participants, all hybrid optimized crystal structures (apart from compound II) have the smallest calculated root mean squared deviations from the experimentally observed structures. It is predicted that a new polymorph of compound V exists under pressure.
    • RGD-Binding Integrins in Head and Neck Cancers

      Ahmedah, H.T.; Patterson, Laurence H.; Shnyder, Steven D.; Sheldrake, Helen M. (2017-05-26)
      Alterations in integrin expression and function promote tumour growth, invasion, metastasis and neoangiogenesis. Head and neck cancers are highly vascular tumours with a tendency to metastasise. They express a wide range of integrin receptors. Expression of the αv and β1 subunits has been explored relatively extensively and linked to tumour progression and metastasis. Individual receptors αvβ3 and αvβ5 have proved popular targets for diagnostic and therapeutic agents but lesser studied receptors, such as αvβ6, αvβ8, and β1 subfamily members, also show promise. This review presents the current knowledge of integrin expression and function in squamous cell carcinoma of the head and neck (HNSCC), with a particular focus on the RGD-binding integrins, in order to highlight the potential of integrins as targets for personalised tumour specific identification and therapy.
    • Rheged: An Early Historic Kingdom Near the Solway

      McCarthy, Michael R. (2003)
      Rheged has been well known to historians for some time, but it is usually considered from the standpoint of the written sources. This paper seeks to begin the process of wider examination, firstly by discussing salient aspects of the archaeological setting, specifically the Iron Age and Roman background. Secondly, attention is drawn to those elements of the archaeological and written record relating to the location of Rheged, as well as to kingship and power. Earlier assumptions as to the location of Rheged are challenged, and it is suggested that its focus was in the Rhinns of Galloway. By the late sixth century Rheged, led by its great king Urien, was in existence, but it proved to be transient, and within a century or so of the earliest references in the literature, it had become absorbed into the expanding kingdom of Northumbria. Later, the Men of the North provided the heroic ancestry and models appropriate to kings in Wales, and ultimately found a place in one of the most enduring themes in medieval romantic literature.
    • RHX Dating: measurement of the Activation Energy of Rehydroxylation for Fired-Clay Ceramics

      Clelland, Sarah-Jane; Wilson, M.A.; Carter, M.A.; Batt, Catherine M. (2015-04)
      In rehydroxylation (RHX) dating, the activation energy of the rehydroxylation reaction is required first in the estimate of a material's effective lifetime temperature (ELT), and second to correct the RHX rate constant obtained at a given measurement temperature to that at the ELT. Measurement of the activation energy is thus integral to the RHX methodology. In this paper, we report a temperature-step method for the measurement of activation energy and develop fully the underlying theoretical basis. In contrast to obtaining the activation energy from a series of separate experiments (each of which requires the sample to be dehydroxylated prior to measuring the RHX rate constant), the temperature-step method not only requires a single dehydroxylation at 500°C but also eliminates repeated acquisition of Stage I data, which are not required for dating purposes. Since the first temperature step is set to correspond to the temperature at which a dating determination is carried out, the measurement of rate constants at higher temperatures simply becomes an extension of dating. Consequently, the logistics of obtaining the activation energy of rehydroxylation are greatly simplified.
    • The Right to Self–determination and Individual rights in the Era of Decolonization in Sub-Saharan Africa: The Case of UNESCO.

      Morvaridi, Behrooz (2011-12)
      This paper examines the conceptual origins of individual rights that shaped the UN and UNESCO model of human rights and the origins of group rights as they emerged in the post–colonial era to challenge inequality. It argues that the idea of rights to self determination, associated initially with decolonization in Africa based on equal statehood status in international relations, has, since decolonization, reinvigorated the promotion of group or peoples’ rights as a framework for challenging poverty and inequality, including access or rights to development.
    • Riluzole elevates GLT-1 activity and levels in striatal astrocytes

      Carbone, M.; Duty, S.; Rattray, Marcus (2012)
      Drugs which upregulate astrocyte glutamate transport may be useful neuroprotective compounds by preventing excitotoxicity. We set up a new system to identify potential neuroprotective drugs which act through GLT-1. Primary mouse striatal astrocytes grown in the presence of the growth-factor supplement G5 express high levels of the functional glutamate transporter, GLT-1 (also known as EAAT2) as assessed by Western blotting and (3)H-glutamate uptake assay, and levels decline following growth factor withdrawal. The GLT-1 transcriptional enhancer dexamethasone (0.1 or 1 muM) was able to prevent loss of GLT-1 levels and activity following growth factor withdrawal. In contrast, ceftriaxone, a compound previously reported to enhance GLT-1 expression, failed to regulate GLT-1 in this system. The neuroprotective compound riluzole (100 muM) upregulated GLT-1 levels and activity, through a mechanism that was not dependent on blockade of voltage-sensitive ion channels, since zonasimide (1 mM) did not regulate GLT-1. Finally, CDP-choline (10 muM-1 mM), a compound which promotes association of GLT-1/EAAT2 with lipid rafts was unable to prevent GLT-1 loss under these conditions. This observation extends the known pharmacological actions of riluzole, and suggests that this compound may exert its neuroprotective effects through an astrocyte-dependent mechanism.
    • Riluzole neuroprotection in a parkinson’s disease model involves suppression of reactive astrocytosis but not GLT-1 regulation.

      Carbone, M.; Duty, S.; Rattray, Marcus (2012-04)
      Background: Riluzole is a neuroprotective drug used in the treatment of motor neurone disease. Recent evidence suggests that riluzole can up-regulate the expression and activity of the astrocyte glutamate transporter, GLT-1. Given that regulation of glutamate transport is predicted to be neuroprotective in Parkinson’s disease, we tested the effect of riluzole in parkinsonian rats which had received a unilateral 6-hydroxydopamine injection into the median forebrain bundle. Results: Rats were treated with intraperitoneal riluzole (4 mg/kg or 8 mg/kg), 1 hour before the lesion then once daily for seven days. Riluzole produced a modest but significant attenuation of dopamine neurone degeneration, assessed by suppression of amphetamine-induced rotations, preservation of tyrosine hydroxylase positive neuronal cell bodies in the substantia nigra pars compacta and attenuation of striatal tyrosine hydroxylase protein loss. Seven days after 6-hydroxydopamine lesion, reactive astrocytosis was observed in the striatum, as determined by increases in expression of glial fibrillary acidic protein, however the glutamate transporter, GLT-1, which is also expressed in astrocytes was not regulated by the lesion. Conclusions: The results confirm that riluzole is a neuroprotective agent in a rodent model of parkinson’s disease. Riluzole administration did not regulate GLT-1 levels but significantly reduced GFAP levels, in the lesioned striatum. Riluzole suppression of reactive astrocytosis is an intriguing finding which might contribute to the neuroprotective effects of this drug.
    • Riluzole–Triazole Hybrids as Novel Chemical Probes for Neuroprotection in Amyotrophic Lateral Sclerosis

      Sweeney, J.B.; Rattray, Marcus; Pugh, V.; Powell, L.A. (2018-06-14)
      Despite intense attention from biomedical and chemical researchers, there are few approved treatments for amyotrophic lateral sclerosis (ALS), with only riluzole (Rilutek) and edaravone (Radicava) currently available to patients. Moreover, the mechanistic basis of the activity of these drugs is currently not well-defined, limiting the ability to design new medicines for ALS. This Letter describes the synthesis of triazole-containing riluzole analogues, and their testing in a novel neuroprotective assay. Seven compounds were identified as having neuroprotective activity, with two compounds having similar activity to riluzole.
    • RNA interference by single- and double-stranded siRNA with a DNA extension containing a 3' nuclease-resistant mini-hairpin structure

      Allison, Simon J.; Milner, J. (2014)
      Selective gene silencing by RNA interference (RNAi) involves double-stranded small interfering RNA (ds siRNA) composed of single-stranded (ss) guide and passenger RNAs. siRNA is recognized and processed by Ago2 and C3PO, endonucleases of the RNA-induced silencing complex (RISC). RISC cleaves passenger RNA, exposing the guide RNA for base-pairing with its homologous mRNA target. Remarkably, the 3′ end of passenger RNA can accommodate a DNA extension of 19-nucleotides without loss of RNAi function. This construct is termed passenger-3′-DNA/ds siRNA and includes a 3′-nuclease-resistant mini-hairpin structure. To test this novel modification further, we have now compared the following constructs: (I) guide-3′-DNA/ds siRNA, (II) passenger-3′-DNA/ds siRNA, (III) guide-3′-DNA/ss siRNA, and (IV) passenger-3′-DNA/ss siRNA. The RNAi target was SIRT1, a cancer-specific survival factor. Constructs I–III each induced selective knock-down of SIRT1 mRNA and protein in both noncancer and cancer cells, accompanied by apoptotic cell death in the cancer cells. Construct IV, which lacks the SIRT1 guide strand, had no effect. Importantly, the 3′-DNA mini-hairpin conferred nuclease resistance to constructs I and II. Resistance required the double-stranded RNA structure since single-stranded guide-3′-DNA/ss siRNA (construct III) was susceptible to serum nucleases with associated loss of RNAi activity. The potential applications of 3′-DNA/siRNA constructs are discussed.
    • Road my body goes: re-creating ancestors from stone at the great moai quarry of Rano Raraku, Rapa Nui (Easter Island)

      Richards, C.; Croucher, Karina T.; Paoa, T.; Parish, T.; Tucki, E.; Welham, K. (2011)
      Recognizable throughout the world, the stone statues (moai) of Rapa Nui (Easter Island) represent the largest monolithic architecture produced in Polynesia. The exquisitely carved and finished head and torso of each statue testifies to a skill in stone carving and dressing unmatched throughout the Pacific. Yet, approximately one thousand ‘classic’ statues were produced at the quarries within a few hundred years. What was the ritual status of the quarry and the labour necessary to produce the numbers of statues that allowed Heyerdahl to declare that the ‘whole mountain massif has been reshaped, the volcano has been greedily cut up’ (1958: 83)? What was it like to go to work at Rano Raraku? By drawing on a range of evidence we argue that walking to and labouring at Rano Raraku represented a spatial and temporal journey to a place of highly dangerous forces, a cosmogonic centre where prehistoric Rapa Nui people came face to face with their ancestors and the Polynesian gods.
    • A Robust and Reliable Test to Measure Stereopsis in the Clinic

      Hess, R.F.; Ding, R.; Clavagnier, S.; Liu, C.; Guo, C.; Viner, Catherine; Barrett, Brendan T.; Radia, Krupali; Zhou, J. (2016-03)
      Purpose: The purpose of this study was to develop a convenient test of stereopsis in the clinic that is both robust and reliable and capable of providing a measure of variability necessary to make valid comparisons between measurements obtained at different occasions or under different conditions. Methods: Stereo acuity was measured based on principles derived from the laboratory measurement of stereopsis (i.e., staircase method). Potential premeasurement compensations are described if there is a significant degree of ocular misalignment, reduced visual acuity, or aniseikonia. Forty-six adults at McGill University, 44 adults at Auckland University, and 51 adults from the University of Bradford, with an age range of 20 to 65 years old and normal or corrected-to-normal vision participated in this study. Results: Stereo acuity within this normal population was widely distributed, with a significant percentage (28%) of the population with only coarse stereo (>300 arc seconds). Across subjects, the SD was approximately 25% of the mean. Measurements at two different times were strongly (r = 0.79) and significantly (P < 0.001) correlated, with little to no significant (P = 0.79) bias (0.01) between test and retest measures of stereopsis. Conclusions: The application enables measurements over the wide disparity range and not just at the finest disparities. In addition, it allows changes in stereopsis of the order of 1.9 to be statistically distinguished.
    • Rod Electroretinograms Elicited by Silent Substitution Stimuli from the Light-Adapted Human Eye

      Maguire, John; Parry, Neil R.A.; Kremers, Jan; Kommanapalli, Deepika; Murray, I.J.; McKeefry, Declan J. (2016-08)
      Purpose: To demonstrate that silent substitution stimuli can be used to generate electroretinograms (ERGs) that effectively isolate rod photoreceptor function in humans without the need for dark adaptation, and that this approach constitutes a viable alternative to current clinical standard testing protocols. Methods: Rod-isolating and non-isolating sinusoidal flicker stimuli were generated on a 4 primary light-emitting diode (LED) Ganzfeld stimulator to elicit ERGs from participants with normal and compromised rod function who had not undergone dark-adaptation. Responses were subjected to Fourier analysis, and the amplitude and phase of the fundamental were used to examine temporal frequency and retinal illuminance response characteristics. Results: Electroretinograms elicited by rod-isolating silent substitution stimuli exhibit low-pass temporal frequency response characteristics with an upper response limit of 30 Hz. Responses are optimal between 5 and 8 Hz and between 10 and 100 photopic trolands (Td). There is a significant correlation between the response amplitudes obtained with the silent substitution method and current standard clinical protocols. Analysis of signal-to-noise ratios reveals significant differences between subjects with normal and compromised rod function. Conclusions: Silent substitution provides an effective method for the isolation of human rod photoreceptor function in subjects with normal as well as compromised rod function when stimuli are used within appropriate parameter ranges. Translational Relevance: This method of generating rod-mediated ERGs can be achieved without time-consuming periods of dark adaptation, provides improved isolation of rod- from cone-based activity, and will lead to the development of faster clinical electrophysiologic testing protocols with improved selectivity.
    • Role of 5-HT receptor mechanisms in sub-chronic PCP-induced reversal learning deficits in the rat

      McLean, Samantha L.; Woolley, M.L.; Thomas, D.; Neill, Joanna C. (2009-10-15)
      Rationale: 5-HT receptor mechanisms have been suggested to mediate improvements in cognition in schizophrenia. Aim: To investigate the involvement of 5-HT receptor mechanisms in sub-chronic PCP-induced reversal learning deficits in female rats, a task of relevance to schizophrenia. Methods: Adult female hooded-Lister rats were trained to perform an operant reversal learning task and then received sub-chronic PCP (2 mg/kg) or vehicle i.p. twice daily for seven days, followed by 7-days washout. Rats then received an acute dose of the 5-HT7 receptor antagonist SB-269970A (1.0, 3.0, 10.0 mg/kg; i.p.) or vehicle. In experiment 2, PCP-treated rats received the selective 5-HT2C receptor antagonist, SB-243213A acutely (1.0, 3.0, 10.0 mg/kg; i.p.) or vehicle. In experiment 3, PCP-treated rats received the 5-HT1A receptor partial agonist, buspirone (0.15625, 0.3125, 0.625 mg/kg, i.p.) in combination with the selective 5-HT1A receptor antagonist WAY-100635 (0.3, 1.0 mg/kg). Results: In all experiments sub-chronic PCP significantly impaired reversal phase performance (P<0.01-0.001), with no effect in the initial phase. SB-269970A at 3.0 and 10.0 mg/kg significantly improved the PCP-induced deficit (P<0.05). SB-243213A also significantly attenuated the deficit at 10 mg/kg (P<0.05). In experiment 3, buspirone attenuated the deficit with significant effects at 0.3125 mg/kg and 0.625 mg/kg (P<0.05). WAY-100635 at 0.3 and 1.0 mg/kg produced a partial attenuation of buspirone’s effect as buspirone (0.3125 mg/kg) in the presence of WAY-100635 did not significantly reverse the PCP-induced deficit. Conclusions: These studies implicate the role of 5-HT7, 5-HT2C and 5-HT1A receptors in the improvement of cognitive dysfunction of relevance to schizophrenia.
    • The role of constitutive pka-mediated phosphorylation in the regulation of basal ICa in isolated rat cardiac myocytes.

      Bracken, N.; El-Kadri, M.; Hart, G.; Hussain, Munir (2006)
      1 Pharmacological inhibitors of protein kinase A (PKA) and protein phosphatases 1/2A were used to determine whether basal L-type Ca2+ current (ICa) observed in the absence of exogenous ß-adrenergic receptor stimulation is sustained by PKA-mediated phosphorylation. Amphotericin B was used to record whole-cell ICa in the perforated patch-clamp configuration. 2 Calyculin A and isoprenaline (both 1 ¿mol l¿1) increased basal ICa (P<0.05), whereas H-89 inhibited ICa in a concentration-dependent manner with an IC50 ~5 ¿mol l¿1. H-89 also inhibited the response to 1.0 ¿mol l¿1 isoprenaline, although relatively high concentrations (30 ¿mol l¿1) were required to achieve complete suppression of the response. 3 Double-pulse protocols were used to study the effects of 10 ¿mol l¿1 H-89 on time-dependent recovery of ICa from voltage-dependent inactivation as well as the steady-state gating of ICa. T0.5 (time for ICa to recover to 50% of the preinactivation amplitude) increased in the presence of H-89 (P<0.05) but was unaffected by calyculin A or isoprenaline. 4 Steady-state activation/inactivation properties of ICa were unaffected by 10 ¿mol l¿1 H-89 or 1 ¿mol l¿1 calyculin A, whereas isoprenaline caused a leftward shift in both curves so that V0.5 for activation and inactivation became more negative. 5 Data show that basal ICa is regulated by cAMP-PKA-mediated phosphorylation in the absence of externally applied ß-receptor agonists and that relatively high concentrations of H-89 are required to fully suppress the response to ß-adrenergic receptor stimulation, thereby limiting the value of H-89 as a useful tool in dissecting signalling pathways in intact myocytes.
    • The role of endothelial cell reactive antibodies in peripheral arterial disease

      Lindsey, Nigel J.; Armitage, J.D.; Homer-Vanniasinkam, Shervanthi (2006)
      Objectives It is hypothesised that endothelial cell reactive antibodies (ECRA) play a role in the progression of PAD through activation of endothelial cells and the release of inflammatory cytokines. We aimed to test this hypothesis by assessing levels of ECRA, E-selectin and IL-6 in patients with PAD of varying severity in a case controlled study. Design, materials, methods Patients were assessed clinically and with ankle¿brachial pressure indices. Patients with critical ischaemia (CI, n=30), stable claudicants (SC, n=30), and age-matched controls (AMC, n=20) were studied. Antibody, E-selectin and IL-6 levels were measured using ELISA. Results ECRA levels were significantly raised in the CI group over AMC. IL-6 levels were significantly elevated in both SC and CI over the control group and in CI over SC. There were no significant differences in E-selectin levels between the AMC, SC and CI. Conclusion Our findings support the hypothesis that autoantibodies play a role in promoting PAD by elevating IL-6. The absence of an elevation in E-selectin in this study may be due to its short half-life, and casts doubt on its value as a marker of inflammation in atherosclerosis.
    • The role of ERK5 in endothelial cell function

      Nithianandarajah-Jones, G.N.; Wilm, B.; Goldring, C.E.; Muller, Jurgen; Cross, M.J. (2014-12-01)
      Extracellular-signal-regulated kinase 5 (ERK5), also termed big MAPK1 (BMK1), is the most recently discovered member of the mitogen-activated protein kinase (MAPK) family. It is expressed in a variety of tissues and is activated by a range of growth factors, cytokines and cellular stresses. Targeted deletion of Erk5 in mice has revealed that the ERK5 signalling cascade is critical for normal cardiovascular development and vascular integrity. In vitro studies have revealed that, in endothelial cells, ERK5 is required for preventing apoptosis, mediating shear-stress signalling and regulating tumour angiogenesis. The present review focuses on our current understanding of the role of ERK5 in regulating endothelial cell function.
    • The role of HOXB9 and miR-196a in head and neck squamous cell carcinoma

      Darda, L.; Hakami, F.; Morgan, Richard; Murdoch, C.; Lambert, D.W.; Hunter, K.D. (2015-04-10)
      Background - Previous studies have demonstrated that a number of HOX genes, a family of transcription factors with key roles in early development, are up-regulated in head and neck squamous cell carcinoma (HNSCC) and other cancers. The loci of several Homeobox (HOX) genes also contain microRNAs (miRs), including miR-196a. Methods - Global miR expression and expression of all 39 HOX genes in normal oral keratinocytes (NOKs), oral pre-malignant (OPM) and HNSCC cells was assessed by expression microarray and qPCR and in tissues by immunohistochemistry (IHC) and qPCR of laser microdissected (LCM) tissues. Expression of miR196a and HOXB9 was reduced using anti-miR-196a and siRNA, respectively. Expression microarray profiles of anti-miR196a and pre-miR196a transfected cells were compared to parental cells in order to identify novel targets of miR- 196a. Putative miR196a targets were validated by qPCR and were confirmed as binding to the 3’UTR of miR196a by a dual luciferase reporter assay combined with mutational analysis of the miR-196a binding site. Results - miR-196a and HOXB9 are highly expressed in HNSCC compared to NOKs, a pattern also seen in HNSCC tissues by HOXB9 IHC and qPCR of miR-196a in LCM tissue. Knock-down of miR-196a expression decreased HNSCC cell migration, invasion and adhesion to fibronectin, but had no effect on proliferation. Furthermore, knock-down of HOXB9 expression decreased migration, invasion and proliferation but did not alter adhesion. We identified a novel primary mRNA transcript containing HOXB9 and miR196a-1 as predicted from in-silico analysis. Expression array analysis identified a number of miR196a targets, including MAMDC2 and HOXC8. We confirmed that MAMDC2 is a novel miR-196a target using a dual luciferase reporter assay with the effect abolished on mutation of the binding site. Conclusions - These results show that miR-196a and HOXB9 are overexpressed, perhaps co-ordinately, as HNSCC develops and exert a pro-tumourigenic phenotype in HNSCC and OPM cells.
    • The role of infant life histories in the construction of identities in death: An incremental isotope study of dietary and physiological status among children afforded differential burial

      Craig-Atkins, E.; Towers, Jacqueline R.; Beaumont, Julia (2018-11)
      Objectives Isotope ratio analyses of dentine collagen were used to characterize short-term changes in physiological status (both dietary status and biological stress) across the life course of children afforded special funerary treatment. Materials and Methods Temporal sequences of δ15N and δ13C isotope profiles for incrementally-forming dentine collagen were obtained from deciduous teeth of 86 children from four early-medieval English cemeteries. Thirty-one were interred in child-specific burial clusters, and the remainder alongside adults in other areas of the cemetery. Isotope profiles were categorized into four distinct patterns of dietary and health status between the final prenatal months and death. Results Isotope profiles from individuals from the burial clusters were significantly less likely to reflect weaning curves, suggesting distinctive breastfeeding and weaning experiences. This relationship was not simply a factor of differential age at death between cohorts. There was no association of burial location with stage of weaning at death, nor with isotopic evidence of physiological stress at the end of life. Discussion This study is the first to identify a relationship between the extent of breastfeeding and the provision of child-specific funerary rites. Limited breastfeeding may indicate the mother had died during or soon after birth, or that either mother or child was unable to feed due to illness. Children who were not breastfed will have experienced a significantly higher risk of malnutrition, undernutrition and infection. These sickly and perhaps motherless children received care to nourish them during early life, and were similarly provided with special treatment in death.