Zbiec-Piekarska R.,Central Forensic Laboratory of the Police |
Spolnicka M.,Central Forensic Laboratory of the Police |
Kupiec T.,Institute of Forensic Research |
Makowska Z.,Central Forensic Laboratory of the Police |
And 5 more authors.
Forensic Science International: Genetics | Year: 2014
Age estimation in forensic investigations may complement the prediction of externally visible characteristics and the inference of biogeographical ancestry, thus allowing a better description of an unknown individual. Multiple CpG sites that show linear correlation between age and degree of DNA methylation have been identified in the human genome, providing a selection of candidates for age prediction. In this study, we optimized an assay based on bisulfite conversion and pyrosequencing of 7 CpG sites located in the ELOVL2 gene. Examination of 303 blood samples collected from individuals aged 2-75 years allowed selection of the most informative site, explaining 83% of variation in age. The final linear regression model included two CpG sites in ELOVL2 and enabled age prediction with R2 = 0.859, prediction error = 6.85 and mean absolute deviation MAD = 5.03. Examination of a testing set of 124 blood samples (MAD = 5.75) showed that 68.5% of samples were correctly predicted, assuming that chronological and predicted ages matched ±7 years. It was found that the ELOVL2 methylation status in bloodstains had not changed significantly after 4 weeks of storage in room temperature conditions. Analysis of 45 bloodstains deposited on tissue paper after 5, 10 and 15 years of storage in room conditions indicated that although a gradual decrease of positive PCR results was observed, the general age prediction success rate remained similar and equaled 60-78%. The obtained results show that the ELOVL2 locus provides a very good source of information about human chronological age based on analysis of blood, including bloodstains, and it may constitute a powerful and reliable predictor in future forensic age estimation models. © 2014 Elsevier Ltd. All rights reserved.
PubMed | Institute of Forensic Research, Medical University of Warsaw, Central Forensic Laboratory of the Police, Jagiellonian University and BioVectis
Type: | Journal: Forensic science international. Genetics | Year: 2015
Forensic DNA phenotyping needs to be supplemented with age prediction to become a relevant source of information on human appearance. Recent progress in analysis of the human methylome has enabled selection of multiple candidate loci showing linear correlation with chronological age. Practical application in forensic science depends on successful validation of these potential age predictors. In this study, eight DNA methylation candidate loci were analysed using convenient and reliable pyrosequencing technology. A total number of 41 CpG sites was investigated in 420 samples collected from men and women aged from 2 to 75 years. The study confirmed correlation of all the investigated markers with human age. The five most significantly correlated CpG sites in ELOVL2 on 6p24.2, C1orf132 on 1q32.2, TRIM59 on 3q25.33, KLF14 on 7q32.3 and FHL2 on 2q12.2 were chosen to build a prediction model. This restriction allowed the technical analysis to be simplified without lowering the prediction accuracy significantly. Model parameters for a discovery set of 300 samples were R(2)=0.94 and the standard error of the estimate=4.5 years. An independent set of 120 samples was used to test the model performance. Mean absolute deviation for this testing set was 3.9 years. The number of correct predictions 5 years achieved a very high level of 86.7% in the age category 2-19 and gradually decreased to 50% in the age category 60-75. The prediction model was deterministic for individuals belonging to these two extreme age categories. The developed method was implemented in a freely available online age prediction calculator.
Strawa K.,Medical University of Warsaw |
Markowska A.,Medical University of Warsaw |
Miskiewicz P.,Medical University of Warsaw |
Kus A.,Medical University of Warsaw |
And 7 more authors.
Clinical Endocrinology | Year: 2014
Background T-cell receptor rearrangement excision circles (TREC) are circular DNA molecules generated during T-cell maturation in the thymus. Recent studies suggested that a decreased TREC concentration in peripheral blood may be a general feature of autoimmunity. Our purpose was to assess the TREC concentration in Graves' disease (GD).Methods TREC concentration was assessed by real time PCR in DNA samples isolated from peripheral blood leucocytes among younger (n = 94, age range 6-29 years) and older patients with GD (n = 93, age range 57-80 years) and age-matched controls (n = 206).Results TREC concentration decreased with age in all subjects, but it was significantly higher in GD compared with controls (P = 9·4 × 10-10). TREC concentration was higher (P = 0·0038) in hyperthyroid (n = 78) than euthyroid (n = 82) patients with GD, but in both groups, it remained increased relative to controls (P = 2·2 × 10-11 and P = 4·4 ×10-7, respectively).Conclusions Patients with GD, particularly those with hyperthyroidism, have increased concentration of TREC which may suggest increased rather than decreased thymic activity. Thus, GD does not follow the paradigm suggested for other autoimmune disorders which links autoimmunity with thymic senescence. © 2014 John Wiley & Sons Ltd.
Spolnicka M.,Central Forensic Laboratory of the Police |
Piekarska R.Z.,Central Forensic Laboratory of the Police |
Jaskula E.,Polish Academy of Sciences |
Jaskula E.,Lower Silesian Center for Cellular Transplantation with National Bone Marrow Donor Registry |
And 13 more authors.
Clinical Epigenetics | Year: 2016
Background: Our recent study demonstrated that DNA methylation status in a set of CpGs located in ELOVL2, C1orf132, TRIM59, KLF14, and FHL2 can accurately predict calendar age in blood. In the present work, we used these markers to evaluate the effect of allogeneic hematopoietic stem cell transplantation (HSCT) on the age-related methylation signature of human blood. Methods: DNA methylation in 32 CpGs was investigated in 16 donor-recipient pairs using pyrosequencing. DNA was isolated from the whole blood collected from recipients 27–360 days (mean 126) after HSCT and from the donors shortly before the HSCT. Results: It was found that in the recipients, the predicted age did not correlate with their calendar age but was correlated with the calendar age (r = 0.94, p = 4 × 10−8) and predicted age (r = 0.97, p = 5 × 10−10) of a respective donor. Despite this strong correlation, the predicted age of a recipient was consistently lower than the predicted age of a donor by 3.7 years (p = 7.8 × 10−4). This shift was caused by hypermethylation of the C1orf132 CpGs, for C1orf132 CpG_1. Intriguingly, the recipient-donor methylation difference correlated with calendar age of the donor (r = 0.76, p = 6 × 10−4). This finding could not trivially be explained by shifts of the major cellular factions of blood. Conclusions: We confirm the single previous report that after HSCT, the age of the donor is the major determinant of age-specific methylation signature in recipient’s blood. A novel finding is the unique methylation dynamics of C1orf132 which encodes MIR29B2C implicated in the self-renewing of hematopoietic stem cells. This observation suggests that C1orf132 could influence graft function after HSCT. © 2016, The Author(s).
PubMed | Jagiellonian University, Medical University of Warsaw, Polish Academy of Sciences, Central Forensic Laboratory of the Police and Medical University of Lódz
Type: | Journal: Clinical epigenetics | Year: 2016
Kowalczuk A.P.,Polish National Medicines Institute |
Lozak A.,Polish National Medicines Institute |
Bachlinski R.,Central Forensic Laboratory of the Police |
Duszynska A.,Central Forensic Laboratory of the Police |
And 2 more authors.
Acta Poloniae Pharmaceutica - Drug Research | Year: 2015
Psychotria viridis (chacruna) is a hallucinogenic plant with psychoactive properties associated with the presence of N,N-dimethyltryptamine (DMT). This species is primarily known as an ingredient of the beverage Ayahuasca, but dry leaves are also smoked by recreational users. The plant is controlled in Poland and France and its proper identification poses many challenges due to the fact that genus Psychotria is relatively large and there are other species that are easily confused with chacruna. The aim of the present work was to develop an effective authentication procedure for the dried and shredded leaves of P. viridis, to be used in comparison of chemical and botanical characteristics of its commercial products. Dried leaves of P. viridis originating from Brazil, Peru and Hawaii were purchased from Internet providers. For DMT identification, thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) methods have been elaborated, validated and applied. In order to clarify the existing differences among samples, chemometric methods have been used. Botanical features and the gas chromatography tandem mass spectrometry (GC-MS) chromatograms have been analyzed using hierarchical cluster analysis (HCA). Our studies revealed significant variety among plant material marketed as P. viridis. Grouping of samples based on their micromorphology features and GC-MS results did not correspond well with the presence of DMT. Based on our results an indisputable identification of dried specimens as P. viridis is very problematic. It is necessary to postulate changes in legislation regarding regulation of P. viridis and replace it with DMT as controlled substance. © 2015, Polish Pharmaceutical Society. All rights reserved.
Klemczak K.,Central Forensic Laboratory of the Police |
Szczepanski T.M.,Central Forensic Laboratory of the Police |
Wieckiewicz U.,Central Forensic Laboratory of the Police |
Kulczyk T.,Ministry of Interior
Journal of Forensic Sciences | Year: 2015
This paper presents the case of a cadaver buried in a pine grove, in loamy soil at a depth of about 1.8 m. The case is quite interesting due to the fact that one hand of the victim was bare and the other one was clad in a latex glove. In the laboratory, the severed hands were gently washed with water. The left palm exhibited signs of advanced decomposition. The ridges were visible only on the thumb. The gloved hand was found to be in much better condition. There were signs of saponification on the palm. Ridges on both the palm and all fingers were visible. The fingers of the right hand, palm prints, and the thumb of the left hand were successfully matched against a ten-print card. The match provided the police with a positive identification of the missing person. © 2014 American Academy of Forensic Sciences.