Development and optimization of the VISAGE basic prototype tool for forensic age estimation

The VISAGE (VISible Attributes through GEnomics) consortium aims to develop, optimize and validate prototype tools to broaden the use of DNA intelligence methods in forensic routine laboratories. This includes age estimation based on the quantification of DNA methylation at specific CpG sites. Here, we present the VISAGE basic prototype tool for age estimation targeting 32 CpGs from five genes ELOVL

Development of an epigenetic age predictor for costal cartilage with a simultaneous somatic tissue differentiation system

Age prediction from DNA has been a topic of interest in recent years due to the promising results obtained when using epigenetic markers. Since DNA methylation gradually changes across the individual's lifetime, prediction models have been developed accordingly for age estimation. The tissue-dependence for this biomarker usually necessitates the development of tissue-specific age prediction models, in this way, multiple models for age inference have been constructed for the most commonly encountered forensic tissues (blood, oral mucosa, semen). The analysis of skeletal remains has also been attempted and prediction models for bone have now been reported. Recently, the VISAGE Enhanced Tool was developed for the simultaneous DNA methylation analysis of 8 age-correlated loci using targeted high-throughput sequencing. It has been shown that this method is compatible with epigenetic age estimation models for blood, buccal cells, and bone. Since when dealing with decomposed cadavers or postmortem samples, cartilage samples are also an important biological source, an age prediction model for cartilage has been generated in the present study based on methylation data collected using the VISAGE Enhanced Tool. In this way, we have developed a forensic cartilage age prediction model using a training set composed of 109 samples (19–74 age range) based on DNA methylation levels from three CpGs in FHL2, TRIM59 and KLF14, using multivariate quantile regression which provides a mean absolute error (MAE) of ± 4.41 years. An independent testing set composed of 72 samples (19–75 age range) was also analyzed and provided an MAE of ± 4.26 years. In addition, we demonstrate that the 8 VISAGE markers, comprising EDARADD, TRIM59, ELOVL2, MIR29B2CHG, PDE4C, ASPA, FHL2 and KLF14, can be used as tissue prediction markers which provide reliable blood, buccal cells, bone, and cartilage differentiation using a developed multinomial logistic regression model. A training set composed of 392 samples (n = 87 blood, n = 86 buccal cells, n = 110 bone and n = 109 cartilage) was used for building the model (correct classifications: 98.72%, sensitivity: 0.988, specificity: 0.996) and validation was performed using a testing set composed of 192 samples (n = 38 blood, n = 36 buccal cells, n = 46 bone and n = 72 cartilage) showing similar predictive success to the training set (correct classifications: 97.4%, sensitivity: 0.968, specificity: 0.991). By developing both a new cartilage age model and a tissue differentiation model, our study significantly expands the use of the VISAGE Enhanced Tool while increasing the amount of DNA methylation-based information obtained from a single sample and a single forensic laboratory analysis. Both models have been placed in the open-access Snipper forensic classification website.</p

Formation of a Bacteriostatic Surface on ZrNb Alloy via Anodization in a Solution Containing Cu Nanoparticles

High strength, excellent corrosion resistance, high biocompatibility, osseointegration ability, and low bacteria adhesion are critical properties of metal implants. Additionally, the implant surface plays a critical role as the cell and bacteria host, and the development of a simultaneously antibacterial and biocompatible implant is still a crucial challenge. Copper nanoparticles (CuNPs) could be a promising alternative to silver in antibacterial surface engineering due to low cell toxicity. In our study, we assessed the biocompatibility and antibacterial properties of a PEO (plasma electrolytic oxidation) coating incorporated with CuNPs (Cu nanoparticles). The structural and chemical parameters of the CuNP and PEO coating were studied with TEM/SEM (Transmission Electron Microscopy/Scanning Electron Microscopy), EDX (Energy-Dispersive X-ray Dpectroscopy), and XRD (X-ray Diffraction) methods. Cell toxicity and bacteria adhesion tests were used to prove the surface safety and antibacterial properties. We can conclude that PEO on a ZrNb alloy in Ca–P solution with CuNPs formed a stable ceramic layer incorporated with Cu nanoparticles. The new surface provided better osteoblast adhesion in all time-points compared with the nontreated metal and showed medium grade antibacterial activities. PEO at 450 V provided better antibacterial properties that are recommended for further investigation

Biocompatibility and Antibacterial Properties of ZnO-Incorporated Anodic Oxide Coatings on TiZrNb Alloy

In a present paper, we demonstrate novel approach to form ceramic coatings with incorporated ZnO nanoparticles (NPs) on low modulus TiZrNb alloy with enhanced biocompatibility and antibacterial parameters. Plasma Electrolytic Oxidation (PEO) was used to integrate ZnO nanoparticles (average size 12–27 nm), mixed with Ca(H2PO2)2 aqueous solution into low modulus TiZrNb alloy surface. The TiZrNb alloys with integrated ZnO NPs successfully showed higher surface porosity and contact angle. XPS investigations showed presence of Ca ions and absence of phosphate ions in the PEO modified layer, what explains higher values of contact angle. Cell culture experiment (U2OS type) confirmed that the surface of as formed oxide-ZnO NPs demonstrated hydrophobic properties, what can affect primary cell attachment. Further investigations showed that Ca ions in the PEO coating stimulated proliferative activity of attached cells, resulting in competitive adhesion between cells and bacteria in clinical situation. Thus, high contact angle and integrated ZnO NPs prevent bacterial adhesion and considerably enhance the antibacterial property of TiZrNb alloys. A new anodic oxide coating with ZnO NPs could be successfully used for modification of low modulus alloys to decrease post-implantation complications

The specific nature of agricultural production in the vicinity of the Czarnorzecko-Strzyżowski Landscape Park / Specyfika produkcji rolniczej na terenach Czarnorzecko-Strzyżowskiego Parku Krajobrazowego

Subject and purpose of work: The aim of the study was to evaluate activities of farms located in the Czarnorzecko-Strzyżowski Landscape Park according to the principles of sustainable development. Materials and methods: Survey research was carried out in the Korczyna commune among randomly selected farm owners at the turn of 2014 and 2015. Results and conclusion: Agricultural production is rather versatile and unspecialized in most of the farm-holdings covered by the study in Korczyna Local Government Area (LGA). Cereals dominate the cropping system, while animal husbandry is predominantly by using the Simmental cattle breed. The farmers are aware that they farm in specific conditions, where sustainable land use is crucial for protecting biodiversity and preserving cultural heritage, thus permitting inhabitants to live in an uncontaminated environment. The Korczyna Local Government Area enjoys favourable conditions for the running of ecological farms, but with farmers visibly lacking in motivation as they consider such farming system as unprofitable, besides requiring lots of time and work.Przedmiot i cel pracy: Celem badań była ocena funkcjonowania gospodarstw rolnych położonych na terenach Czarnorzecko-Strzyżowskiego Parku Krajobrazowego zgodnie z zasadami zrównoważonego rozwoju. Materiały i metody: Badania ankietowe zostały przeprowadzone na terenie gminy Korczyna wśród losowo wybranych właścicieli gospodarstw rolnych na przełomie 2014 i 2015 roku. Wyniki i wnioski: W większości badanych gospodarstw w gminie Korczyna produkcja rolna ma charakter wszechstronny oraz niewyspecjalizowany. W uprawach dominują zboża, a w produkcji zwierzęcej użytkowanie bydła rasy simentalskiej. Rolnicy mają świadomość, że gospodarują w specyficznych warunkach, gdzie zrównoważone użytkowanie gruntów jest bardzo ważne dla ochrony różnorodności biologicznej oraz zachowania dziedzictwa kulturowego, a także pozwala mieszkańcom na życie w nieskażonym środowisku. Gmina Korczyna posiada sprzyjające warunki do tworzenia gospodarstw ekologicznych, ale zaobserwowano brak motywacji ze strony rolników, którzy uważają ten typ gospodarowania za nieopłacalny oraz wymagający dużego nakładu pracy i czasu

The Specific Nature of Agricultural Production in the Vicinity of the Czarnorzecko-Strzyżowski Landscape Park

Subject and purpose of work: The aim of the study was to evaluate activities of farms located in the Czarnorzecko-Strzyżowski Landscape Park according to the principles of sustainable development