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Item Metadata only Adulterants and Contaminants in Psychotropic Herbal Medicines Detected with Mass Spectrometry and Next-Generation DNA Sequencing(Springer, 2018) Hoban, C.L.; Musgrave, I.F.; Coghlan, M.L.; Power, M.W.P.; Byard, R.W.; Nash, C.; Farrington, R.; Maker, G.; Crighton, E.; Trengove, R.; Bunce, M.Introduction: The role of herbal medicine in the treatment of common psychiatric disorders such as anxiety, depression and insomnia has become more established over the past decade. Some herbal preparations such as St John’s wort (Hypericum perforatum) have demonstrated clinical evidence but have also been included in recent reports of widespread adulteration and contamination. Herbal medicines sold in Australia are required to be listed on the Therapeutic Goods Administration’s (TGA) Australian Register of Therapeutic Goods (ARTG) and must comply with strict ingredient and manufacturing guidelines to assure quality and safety. Objective: The aim of this research was to assess whether pharmaceutical adulterants and contaminants were present in psychotropic herbal medicines available in Australia, as a measure of quality, and the effectiveness of regulation. Methods: A two-pronged approach combining next-generation DNA sequencing and small-molecule analysis techniques was undertaken to audit a subset of herbal medicines for the presence of prescription medications, illicit drugs, pesticides, herbicides, heavy metals and contaminant DNA. Small-molecule analysis included liquid chromatography with quadrupole time-of-flight mass spectrometer (LC-QTOF-MS) detection, liquid chromatography with UV/vis diode array (LC-UV) detection, gas chromatography with nitrogen–phosphorus and mass spectrometer detection (GC-NPD/MS) and heavy metal analysis using inductively coupled plasma with mass spectrometer (ICP-MS) detection. Results: In total, 49% (29 of 59) of the investigated herbal medicines had one or more materials not listed on their labels or ARTG registration, including Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES)-listed material (one medicine), heavy metals (12%) or components that could trigger food sensitivity, such as wheat (12%). In contrast to previous studies, no prescription pharmaceutical adulterants were detected, although 10% had undeclared caffeine. Twenty-four percent of herbal medicines had DNA from animal species, including mice and bats, indicating poor quality control. The surveyed herbal medicines included both traditional Chinese medicines (TCM) and Western herbals. Ninety-four percent of TCMs were contaminated or adulterated, compared with 37% of the Western herbals. Only two of the 59 samples contained the listed active ingredient(s) without additional adulterants and contaminants, or missing ingredients. Conclusions: The high levels of contamination found in this study suggests that closer surveillance of herbal medicines is needed in order to assure the required level of quality of herbal medicines available in Australia. The results suggest that the TGA’s low-/high-risk system for regulation coupled with post-market auditing is not keeping unapproved and/or unsafe herbal medicines from the market.Item Open Access GLA-modified RNA treatment lowers GB3 levels in iPSC-derived cardiomyocytes from Fabry-affected individuals(Elsevier (Cell Press), 2023) ter Huurne, M.; Parker, B.L.; Liu, N.Q.; Qian, E.L.; Vivien, C.; Karavendzas, K.; Mills, R.J.; Saville, J.T.; Abu-Bonsrah, D.; Wise, A.F.; Hudson, J.E.; Talbot, A.S.; Finn, P.F.; Martini, P.G.V.; Fuller, M.; Ricardo, S.D.; Watt, K.I.; Nicholls, K.M.; Porrello, E.R.; Elliott, D.A.Recent studies in non-human model systems have shown therapeutic potential of nucleoside-modified messenger RNA (modRNA) treatments for lysosomal storage diseases. Here, we assessed the efficacy of a modRNA treatment to restore the expression of the galactosidase alpha (GLA), which codes for α-Galactosidase A (α-GAL) enzyme, in a human cardiac model generated from induced pluripotent stem cells (iPSCs) derived from two individuals with Fabry disease. Consistent with the clinical phenotype, cardiomyocytes from iPSCs derived from Fabry-affected individuals showed accumulation of the glycosphingolipid Globotriaosylceramide (GB3), which is an α-galactosidase substrate. Furthermore, the Fabry cardiomyocytes displayed significant upregulation of lysosomal-associated proteins. Upon GLA modRNA treatment, a subset of lysosomal proteins were partially restored to wild-type levels, implying the rescue of the molecular phenotype associated with the Fabry genotype. Importantly, a significant reduction of GB3 levels was observed in GLA modRNA-treated cardiomyocytes, demonstrating that α-GAL enzymatic activity was restored. Together, our results validate the utility of iPSC-derived cardiomyocytes from affected individuals as a model to study disease processes in Fabry disease and the therapeutic potential of GLA modRNA treatment to reduce GB3 accumulation in the heart.Item Open Access A first-in-class pan-lysyl oxidase inhibitor impairs stromal remodeling and enhances gemcitabine response and survival in pancreatic cancer(Springer, 2023) Chitty, J.L.; Yam, M.; Perryman, L.; Parker, A.L.; Skhinas, J.N.; Setargew, Y.F.I.; Mok, E.T.Y.; Tran, E.; Grant, R.D.; Latham, S.L.; Pereira, B.A.; Ritchie, S.C.; Murphy, K.J.; Trpceski, M.; Findlay, A.D.; Melenec, P.; Filipe, E.C.; Nadalini, A.; Velayuthar, S.; Major, G.; et al.The lysyl oxidase family represents a promising target in stromal targeting of solid tumors due to the importance of this family in crosslinking and stabilizing fibrillar collagens and its known role in tumor desmoplasia. Using small-molecule drug-design approaches, we generated and validated PXS-5505, a first-in-class highly selective and potent pan-lysyl oxidase inhibitor. We demonstrate in vitro and in vivo that pan-lysyl oxidase inhibition decreases chemotherapy-induced pancreatic tumor desmoplasia and stiffness, reduces cancer cell invasion and metastasis, improves tumor perfusion and enhances the efficacy of chemotherapy in the autochthonous genetically engineered KPC model, while also demonstrating antifibrotic effects in human patient-derived xenograft models of pancreatic cancer. PXS-5505 is orally bioavailable, safe and effective at inhibiting lysyl oxidase activity in tissues. Our findings present the rationale for progression of a pan-lysyl oxidase inhibitor aimed at eliciting a reduction in stromal matrix to potentiate chemotherapy in pancreatic ductal adenocarcinoma.Item Open Access On the rules of engagement for microRNAs targeting protein coding regions(Oxford University Press, 2023) Sapkota, S.; Pillman, K.A.; Dredge, B.K.; Liu, D.; Bracken, J.M.; Kachooei, S.A.; Chereda, B.; Gregory, P.A.; Bracken, C.P.; Goodall, G.J.MiRNAs post-transcriptionally repress gene expression by binding to mRNA 3 UTRs, but the extent to which they act through protein coding regions (CDS regions) is less well established. MiRNA interaction studies show a substantial proportion of binding occurs in CDS regions, however sequencing studies show much weaker effects on mRNA levels than from 3 UTR interactions, presumably due to competition from the translating ribosome. Consequently, most target prediction algorithms consider only 3 UTR interactions. However, the consequences of CDS interactions may have been underestimated, with the reporting of a novel mode of miRNA-CDS interaction requiring base pairing of the miRNA 3 end, but not the canonical seed site, leading to repression of translation with little effect on mRNA turnover. Using extensive reporter, western blotting and bioinformatic analyses, we confirm that miRNAs can indeed suppress genes through CDS-interaction in special circumstances. However, in contrast to that previously reported, we find repression requires extensive base-pairing, including of the canonical seed, but does not strictly require base pairing of the 3 miRNA terminus and is mediated through reducing mRNA levels. We conclude that suppression of endogenous genes can occur through miRNAs binding to CDS, but the requirement for extensive basepairing likely limits the regulatory impacts to modest effects on a small subset of targets.Item Open Access The Australian Reproductive Genetic Carrier Screening Project (Mackenzie's Mission): Design and Implementation(MDPI AG, 2022) Archibald, A.D.; McClaren, B.J.; Caruana, J.; Tutty, E.; King, E.A.; Halliday, J.L.; Best, S.; Kanga-Parabia, A.; Bennetts, B.H.; Cliffe, C.C.; Madelli, E.O.; Ho, G.; Liebelt, J.; Long, J.C.; Braithwaite, J.; Kennedy, J.; Massie, J.; Emery, J.D.; McGaughran, J.; Marum, J.E.; et al.Reproductive genetic carrier screening (RGCS) provides people with information about their chance of having children with autosomal recessive or X-linked genetic conditions, enabling informed reproductive decision-making. RGCS is recommended to be offered to all couples during preconception or in early pregnancy. However, cost and a lack of awareness may prevent access. To address this, the Australian Government funded Mackenzie’s Mission—the Australian Reproductive Genetic Carrier Screening Project. Mackenzie’s Mission aims to assess the acceptability and feasibility of an easily accessible RGCS program, provided free of charge to the participant. In study Phase 1, implementation needs were mapped, and key study elements were developed. In Phase 2, RGCS is being offered by healthcare providers educated by the study team. Reproductive couples who provide consent are screened for over 1200 genes associated with >750 serious, childhood-onset genetic conditions. Those with an increased chance result are provided comprehensive genetic counseling support. Reproductive couples, recruiting healthcare providers, and study team members are also invited to complete surveys and/or interviews. In Phase 3, a mixed-methods analysis will be undertaken to assess the program outcomes, psychosocial implications and implementation considerations alongside an ongoing bioethical analysis and a health economic evaluation. Findings will inform the implementation of an ethically robust RGCS program.Item Open Access Investigating genetic variants in microRNA regulators of Neurokinin-1 receptor in sudden infant death syndrome(Wiley, 2022) Shaukat, Z.; Byard, R.W.; Vink, R.; Hussain, R.; Ricos, M.G.; Dibbens, L.M.Sudden infant death syndrome (SIDS) occurs more often in male than in female infants, suggesting involvement of the X-chromosome. Histopathological studies have suggested that altered expression of the Neurokinin-1 receptor may also play a role in the pathogenesis of SIDS. It was hypothesised that genetic variants in three X-chromosome- encoded microRNA (miRNA/miR), known to down-regulate expression of the Neurokinin-1 receptor, may contribute to SIDS. Aim: To identify sequence variants in the miRNAs within a study cohort (27 cases of SIDS and 28 controls) and determine if there was a difference in the frequencies in male and female SIDS infants. Methods: Genomic DNA prepared from stored blood spots was amplified and sequenced to identify genetic variants in miR500A, miR500B and miR320D2. Results: No novel variants in the miRNAs were identified in our study cohort. We identified one known single-nucleotide polymorphism (SNP) in miR320D2: rs5907732 G/T, in both cases and controls. No significant difference in the SNP frequency was observed between male and female SIDS cases. Conclusion: This pilot study suggests that sequence variants in three miRNAs do not contribute to the reported higher prevalence of SIDS in male infants and do not contribute to the pathogenesis of SIDS in our cohort.Item Metadata only Determination of ester position in isomeric (O-acyl)-hydroxy fatty acids by ion trap mass spectrometry(Wiley, 2016) Marshall, D.L.; Saville, J.T.; Maccarone, A.T.; Ailuri, R.; Kelso, M.J.; Mitchell, T.W.; Blanksby, S.J.Abstract not availableItem Metadata only Animal and cell culture models for cystic fibrosis: which model is right for your application?(Elsevier, 2021) Mc Carron, A.; Parsons, D.; Donnelley, M.Over the past 30 years, a range of cystic fibrosis (CF) animal models have been generated for research purposes. Different species, including mice, rats, ferrets, rabbits, pigs, sheep, zebrafish, and fruit flies, have all been used to model CF disease. While access to such a variety of animal models is a luxury for any research field, it also complicates the decision-making process when it comes to selecting the right model for an investigation. The purpose of this review is to provide a guide for selecting the most appropriate CF animal model for any given application. In this review, the characteristics and phenotypes of each animal model are described, along with a discussion of the key considerations that must be taken into account when choosing a suitable animal model. Available in vitro systems of CF are also described and can offer a useful alternative to using animal models. Finally, the future of CF animal model generation and its use in research are speculated upon.Item Open Access Evaluation of multiple methods for quantification of glycosaminoglycan biomarkers in newborn dried blood spots from patients with severe and attenuated mucopolysaccharidosis-I(MDPI, 2020) Herbst, Z.M.; Urdaneta, L.; Klein, T.; Fuller, M.; Gelb, M.H.All newborn screening (NBS) for mucopolysaccharidosis-I (MPS-I) is carried out by the measurement of α-iduronidase (IDUA) enzymatic activity in dried blood spots (DBS). The majority of low enzyme results are due to pseudodeficiencies, and studies from the Mayo Clinic have shown that the false positive rate can be greatly reduced by including a second-tier analysis of glycosaminoglycans (GAGs) in DBS as part of NBS. In the present study, we obtained newborn DBS from 13 patients with severe MPS-I and 2 with attenuated phenotypes. These samples were submitted to four different GAG mass spectrometry analyses in a comparative study: (1) internal disaccharide; (2) endogenous disaccharide; (3) Sensi-Pro; (4) Sensi-Pro Lite (a variation of Sensi-Pro with a simplified workflow). Patients with attenuated MPS-I show less GAG elevation than those with severe disease, and all MPS-I patients were separated from the reference range using all four methods. The minimal differential factor (lowest GAG marker level in MPS-I samples divided by highest level in the reference range of 30 random newborns) was about two for internal disaccharide, Sensi-Pro, and Sensi-Pro Lite methods. The endogenous disaccharide was clearly the best method with a minimal differential of 16-fold. This study supports use of second-tier GAG analysis of newborn DBS, especially the endogenous disaccharide method, as part of NBS to reduce the false positive rate.Item Metadata only Gene selection for the Australian Reproductive Genetic Carrier Screening Project ("Mackenzie's Mission")(Springer Nature, 2021) Kirk, E.P.; Ong, R.; Boggs, K.; Hardy, T.; Righetti, S.; Ben, K.; Roscioli, T.; Amor, D.J.; Bakshi, M.; Chung, C.W.T.; Colley, A.; Jamieson, R.; Liebelt, J.; Ma, A.; Pachter, N.; Rajagopalan, S.; Ravine, A.; Wilson, M.; Caruana, J.; Casella, R.; et al.Reproductive genetic carrier screening aims to offer couples information about their chance of having children with certain autosomal recessive and X-linked genetic conditions. We developed a gene list for use in "Mackenzie's Mission", a research project in which 10,000 couples will undergo screening. Criteria for selecting genes were: the condition should be life-limiting or disabling, with childhood onset, such that couples would be likely to take steps to avoid having an affected child; and/or be one for which early diagnosis and intervention would substantially change outcome. Strong evidence for gene-phenotype relationship was required. Candidate genes were identified from OMIM and via review of 23 commercial and published gene lists. Genes were reviewed by 16 clinical geneticists using a standard operating procedure, in a process overseen by a multidisciplinary committee which included clinical geneticists, genetic counselors, an ethicist, a parent of a child with a genetic condition and scientists from diagnostic and research backgrounds. 1300 genes met criteria. Genes associated with non-syndromic deafness and non-syndromic differences of sex development were not included. Our experience has highlighted that gene selection for a carrier screening panel needs to be a dynamic process with ongoing review and refinement.Item Metadata only Prospective study assessing associated factors in patients with colon polyps: interim analysis(Wiley, 2017) Cheng, L.Z.T.P.; Siew, D.C.K.; Ovenden, A.; Burt, A.D.; Singh, R.; Asian Pacific Digestive Week (APDW). The Future in Digestive Diseases (23 Sep 2017 - 26 Sep 2017 : Hong Kong)Item Metadata only Ssb1 and Ssb2 cooperate to regulate mouse hematopoietic stem and progenitor cells by resolving replicative stress(American Society of Hematology, 2017) Shi, W.; Vu, T.; Boucher, D.; Biernacka, A.; Nde, J.; Pandita, R.K.; Straube, J.; Boyle, G.M.; Al-Ejeh, F.; Nag, P.; Jeffery, J.; Harris, J.L.; Bain, A.L.; Grzelak, M.; Skrzypczak, M.; Mitra, A.; Dojer, N.; Crosetto, N.; Cloonan, N.; Becherel, O.J.; et al.Hematopoietic stem and progenitor cells (HSPCs) are vulnerable to endogenous damage and defects in DNA repair can limit their function. The 2 single-stranded DNA (ssDNA) binding proteins SSB1 and SSB2 are crucial regulators of the DNA damage response; however, their overlapping roles during normal physiology are incompletely understood. We generated mice in which both Ssb1 and Ssb2 were constitutively or conditionally deleted. Constitutive Ssb1/Ssb2 double knockout (DKO) caused early embryonic lethality, whereas conditional Ssb1/Ssb2 double knockout (cDKO) in adult mice resulted in acute lethality due to bone marrow failure and intestinal atrophy featuring stem and progenitor cell depletion, a phenotype unexpected from the previously reported single knockout models of Ssb1 or Ssb2 Mechanistically, cDKO HSPCs showed altered replication fork dynamics, massive accumulation of DNA damage, genome-wide double-strand breaks enriched at Ssb-binding regions and CpG islands, together with the accumulation of R-loops and cytosolic ssDNA. Transcriptional profiling of cDKO HSPCs revealed the activation of p53 and interferon (IFN) pathways, which enforced cell cycling in quiescent HSPCs, resulting in their apoptotic death. The rapid cell death phenotype was reproducible in in vitro cultured cDKO-hematopoietic stem cells, which were significantly rescued by nucleotide supplementation or after depletion of p53. Collectively, Ssb1 and Ssb2 control crucial aspects of HSPC function, including proliferation and survival in vivo by resolving replicative stress to maintain genomic stability.Item Metadata only Line of fire – what happened at the Wantabadgery seige?(Humana Press, 2018) Byard, R.; Ford, A.; Raymond, T.; Sofonia, J.; Kaluza, O.; Barnes, D.A gunfight between police and a gang of men led by the self-styled "Captain Moonlite", a.k.a. George Scott, occurred on 16th November 1879 at a farmhouse near Wantabadgery Station in the colony of New South Wales. The skirmish resulted in the deaths of two bushrangers and one police officer. As a result, Captain Moonlite and Thomas Rogan were hung in Sydney's Darlinghurst Gaol on 20 January 1880 for the murder of Constable Edward Webb-Bowen. Culpability for firing the fatal shot, however, has remained a source of controversy. Information obtained from an analysis of historical records was used to guide an archeological excavation at the scene of the shooting in which Terrestrial Laser Scanning (TLS) technology was employed to produce a digital (3D) terrain model of the siege location. Utilizing the terrain model, the relative positions of Moonlite, Webb-Bowen, and the other gang members were established with possible projectile trajectories plotted. This, in combination with inquest evidence from a gun maker and the medical practitioner who examined Constable Webb-Bowen's wound, indicates that the most likely shooter was Gus Warnicke, aged 15 years, the youngest member of the gang, who was also killed in the exchange of fire.Item Metadata only Mucosal-associated invariant T cells are reduced and functionally immature in the peripheral blood of primary Sjögren's syndrome patients(John Wiley & Sons, 2016) Wang, J.; Macardle, C.; Weedon, H.; Beroukas, D.; Banovic, T.The frequencies, immunophenotype, and function of mucosal-associated invariant T (MAIT) cells were studied in patients with primary Sjögren syndrome (pSS) and healthy controls. MAIT cells were significantly decreased in the peripheral blood (PB) of patients with pSS. Vα7.2+ MAIT cells were detected in the salivary gland tissue from pSS patients, but not in controls, indicating that the reduction of MAIT cells in PB might be due to migration into the target tissue. Furthermore, the residual peripheral blood MAIT cells in pSS patients showed altered immunophenotype and function. While MAIT cells from controls were almost exclusively CD8+ and expressed an effector memory immunophenotype, in pSS patients they were enriched in CD4+ and naïve subpopulations. Consistently, the functional studies demonstrated that MAIT cells from pSS showed a lower level of activation with reduced expression of CD69 and CD154 (CD40L), and a lower production of TNF and IFN-γ. In summary, our findings demonstrate that MAIT cells were reduced and phenotypically and functionally altered in PB of pSS patients. The altered function of MAIT cells in target tissues from pSS patients may result in dysregulation of mucosal immunity leading to microbial damage of mucosal surfaces and subsequent initiation of autoimmune response.Item Metadata only Assessing impact of organised breast screening across small residential areas-development and internal validation of a prediction model(Wiley-Blackwell, 2017) Buckley, E.; Farshid, G.; Gill, G.; Kollias, J.; Koczwara, B.; Karapetis, C.; Adams, J.; Joshi, R.; Keefe, D.; Niyonsenga, T.; Powell, K.; Fusco, K.; Eckert, M.; Beckmann, K.; Roder, D.Monitoring screening mammography effects in small areas is often limited by small numbers of deaths and delayed effects. We developed a risk score for breast cancer death to circumvent these limitations. Screening, if effective, would increase post-diagnostic survivals through lead-time and related effects, as well as mortality reductions. Linked cancer and BreastScreen data at four hospitals (n = 2,039) were used to investigate whether screened cases had higher recorded survivals in 13 small areas, using breast cancer deaths as the outcome (M1), and a risk of death score derived from TNM stage, grade, histology type, hormone receptor status, and related variables (M2). M1 indicated lower risk of death in screened cases in 12 of the 13 areas, achieving statistical significance (p < .05) in 5. M2 indicated lower risk scores in screened cases in all 13 areas, achieving statistical significance in 12. For cases recently screened at diagnosis (<6 months), statistically significant reductions applied in 8 areas (M1) and all 13 areas (M2). Screening effects are more detectable in small areas using these risk scores than death itself as the outcome variable. An added advantage is the application of risk scores for providing a marker of screening effect soon after diagnosis.Item Metadata only Toll like receptor 4 activation can be either detrimental or beneficial following mild repetitive traumatic brain injury depending on timing of activation(Mary Ann Liebert, 2016) Corrigan, F.; Vink, R.; 12th Symposium of the International Neurotrauma Society (1 Feb 2016 - 4 Feb 2016 : Cape Town, SOUTH AFRICA)Item Metadata only Toll like receptor 4 activation can be either detrimental or beneficial following mild repetitive traumatic brain injury depending on timing of activation(Elsevier, 2017) Corrigan, F.; Arulsamy, A.; Collins-Praino, L.; Holmes, J.; Vink, R.Abstract not availableItem Metadata only The landscape of somatic mutations in infant MLL-rearranged acute lymphoblastic leukemias(Nature Publishing Group, 2015) Andersson, A.; Ma, J.; Wang, J.; Chen, X.; Gedman, A.; Dang, J.; Nakitandwe, J.; Holmfeldt, L.; Parker, M.; Easton, J.; Huether, R.; Kriwacki, R.; Rusch, M.; Wu, G.; Li, Y.; Mulder, H.; Raimondi, S.; Pounds, S.; Kang, G.; Shi, L.; et al.Infant acute lymphoblastic leukemia (ALL) with MLL rearrangements (MLL-R) represents a distinct leukemia with a poor prognosis. To define its mutational landscape, we performed whole-genome, exome, RNA and targeted DNA sequencing on 65 infants (47 MLL-R and 18 non-MLL-R cases) and 20 older children (MLL-R cases) with leukemia. Our data show that infant MLL-R ALL has one of the lowest frequencies of somatic mutations of any sequenced cancer, with the predominant leukemic clone carrying a mean of 1.3 non-silent mutations. Despite this paucity of mutations, we detected activating mutations in kinase-PI3K-RAS signaling pathway components in 47% of cases. Surprisingly, these mutations were often subclonal and were frequently lost at relapse. In contrast to infant cases, MLL-R leukemia in older children had more somatic mutations (mean of 6.5 mutations/case versus 1.3 mutations/case, P = 7.15 × 10(-5)) and had frequent mutations (45%) in epigenetic regulators, a category of genes that, with the exception of MLL, was rarely mutated in infant MLL-R ALL.Item Metadata only Validation of ultraviolet, infrared, and narrow band light alternate light sources for detection of bruises in a pigskin model(Springer, 2016) Olds, K.; Byard, R.; Winskog, C.; Langlois, N.Alternate light sources such as ultraviolet, narrow band, and infrared have been used in an attempt to reveal the presence of bruising that is not otherwise apparent (inapparent). The following study evaluates the ability of alternate light sources to enhance visibility of bruises by employing an objective assessment of digital photography images in conjunction with histology. A pigskin model was employed with bruises created by injection of blood to be not visible or barely visible (inapparent) under white light. The pigskin was photographed using alternate light source illumination. Images were assessed using the program Fiji® to measure enhancement in terms of bruise length (cm). Photography results were compared with histology to confirm the presence of bruising. Violet and blue light sources produced the greatest enhancement, both with a p < 0.0001. Regions that were not bruises were also enhanced with light sources in this study, indicating that light sources are not specific, and that their use to enhance the visibility of bruising should be undertaken with caution.Item Open Access Inherited coding variants at the CDKN2A locus influence susceptibility to acute lymphoblastic leukaemia in children(Nature Publishing Group, 2015) Xu, H.; Zhang, H.; Yang, W.; Yadav, R.; Morrison, A.; Qian, M.; Devidas, M.; Liu, Y.; Perez-Andreu, V.; Zhao, X.; Gastier-Foster, J.; Lupo, P.; Neale, G.; Raetz, E.; Larsen, E.; Bowman, W.; Carroll, W.; Winick, N.; Williams, R.; Hansen, T.; et al.There is increasing evidence from genome-wide association studies for a strong inherited genetic basis of susceptibility to acute lymphoblastic leukaemia (ALL) in children, yet the effects of protein-coding variants on ALL risk have not been systematically evaluated. Here we show a missense variant in CDKN2A associated with the development of ALL at genome-wide significance (rs3731249, P=9.4 × 10(-23), odds ratio=2.23). Functional studies indicate that this hypomorphic variant results in reduced tumour suppressor function of p16(INK4A), increases the susceptibility to leukaemic transformation of haematopoietic progenitor cells, and is preferentially retained in ALL tumour cells. Resequencing the CDKN2A-CDKN2B locus in 2,407 childhood ALL cases reveals 19 additional putative functional germline variants. These results provide direct functional evidence for the influence of inherited genetic variation on ALL risk, highlighting the important and complex roles of CDKN2A-CDKN2B tumour suppressors in leukaemogenesis.