Paternity analysis based on NGM SElect system in the Medical and Forensic Genetics Laboratory, Department of Forensic Medicine, Medical University of Lodz.

The aim of the study was to evaluate the usefulness of the NGM SElect multiplex kit for paternity testing in the population of central Poland, and compare it with the IDENTIFILER system. The study material consisted of buccal swabs taken from individuals who reported to the Medical and Forensic Genetics Laboratory in Lodz. Samples from 450 trio cases of disputed paternity carried out in 2010-2014 were investigated. Genomic DNA was extracted from buccal swabs collected from 1,350 individuals using the Swab kit (A and A Biotechnology) according to the manufacturer's protocol. DNA amplification was performed using the AmpFℓSTR® NGM SelectTM PCR Amplification Kit (Life Technologies). PCR products were separated by capillary electrophoresis using HID 3500 Genetic Analyzer. In the analyzed cases with paternity confirmation in the NGM SElect system, the maximum value of PI was 3.9 × 1012, which corresponds to the probability of paternity W = 99.9999999999%. It was thus significantly higher than analogical parameters obtained in the IDENTIFILER system (PI = 6.0 × 1010, W = 99.99999999%). The NGM SElect kit was unable to resolve just one case out of 450, which represents only 0.2% of all analyzed disputed paternity cases. The study showed the SE33 (ACTBP2) locus to have the highest evidence value in paternity analysis out of all investigated autosomal STRs.

Titania-Supported Catalysts for Levulinic Acid Hydrogenation: Influence of Support and its Impact on γ-Valerolactone Yield.

A series of titania-supported ruthenium and platinum catalysts was investigated in the levulinic acid hydrogenation towards γ-valerolactone, a key reaction for the catalytic transformation of biomass. It was shown that various morphologies and phases of titania strongly influence the physicochemical and catalytic properties of supported Ru and Pt catalysts in different ways. In the case of the catalyst supported on mixed TiO2 phases, Ru particles are exclusively located on the minority rutile crystallites, whereas such an effect was not observed for platinum. The platinum catalyst activity could be increased when the metal was dispersed on the large surface-area anatase, which was not the case for ruthenium as a result of its agglomeration on this support. The activity of ruthenium on anatase could be increased in two ways: a) when RuO2 formation during catalyst preparation was avoided; b) when pure anatase support material was modified so that it exhibited no microporosity. The obtained results allow a better understanding of the role of the support for Ru and Pt catalysts.

Dangers resulting from DNA profiling of biological materials derived from patients after allogeneic hematopoietic stem cell transplantation (allo-HSCT) with regard to forensic genetic analysis.

The study documents the risk that comes with DNA analysis of materials derived from patients after allogeneic hematopoietic stem cell transplantation (allo-HSCT) in forensic genetics. DNA chimerism was studied in 30 patients after allo-HSCT, based on techniques applied in contemporary forensic genetics, i.e. real-time PCR and multiplex PCR-STR with the use of autosomal DNA as well as Y-DNA markers. The results revealed that the DNA profile of the recipient's blood was identical with the donor's in the majority of cases. Therefore, blood analysis can lead to false conclusions in personal identification as well as kinship analysis. An investigation of buccal swabs revealed a mixture of DNA in the majority of recipients. Consequently, personal identification on the basis of stain analysis of the same origin may be impossible. The safest (but not ideal) material turned out to be the hair root. Its analysis based on autosomal DNA revealed 100% of the recipient's profile. However, an analysis based on Y-chromosome markers performed in female allo-HSCT recipients with male donors demonstrated the presence of donor DNA in hair cells - similarly to the blood and buccal swabs. In the light of potential risks arising from DNA profiling of biological materials derived from persons after allotransplantation in judicial aspects, certain procedures were proposed to eliminate such dangers. The basic procedures include abandoning the approach based exclusively on blood collection, both for kinship analysis and personal identification; asking persons who are to be tested about their history of allo-HSCT before sample collection and profile entry in the DNA database, and verification of DNA profiling based on hair follicles in uncertain cases.

Dynamic Vascular Pattern (DVP), a quantification tool for contrast enhanced ultrasound.

Contrast-enhanced ultrasound (CEUS) is widely applied in tumour diagnosis, especially for focal liver lesions (FLL), due to its high sensitivity and specificity. According to the European Federation of Societies for Ultrasound in Medicine and Biology (EFSUMB) CEUS guidelines (2012) and non-liver guidelines (2011), the majority of tumours, regardless of location, show specific CEUS enhancement patterns that can distinguish benign from malignant lesions. However, even experienced clinicians evaluating FLL may find occasional irregularities in these patterns, due to particular FLL pathologies, that make a definitive diagnosis difficult. Hence, there is a need to train physicians to utilize contrast enhancement kinetics to aid in the correct interpretation of data from CEUS examinations in patients with divergent liver tumour pathologies. Here we report on a CEUS quantitation software, SonoLiver®, to verify and improve diagnostic accuracy in the characterization of suspicious liver lesions through the analysis of dynamic vascular patterns (DVP).

Wolfram syndrome in the Polish population: novel mutations and genotype-phenotype correlation.

OBJECTIVE: Wolfram syndrome is a rare form of diabetes mellitus associated with optic atrophy and disorders of different organs (e.g. diabetes insipidus, hearing loss, ataxia, anaemia and many others). This syndrome is caused by recessive mutations in the wolframin gene (WFS1) localized on chromosome 4p16·1. The aim of this study was to identify the causative mutations in WFS1 in a group of Polish patients with suspected Wolfram syndrome. PATIENTS AND MEASUREMENTS: Nine patients with clinical symptoms consistent with Wolfram syndrome (at least diabetes mellitus and optic atrophy) and 22 first-degree relatives were examined. The molecular analysis was carried out by direct sequencing of the exons, the exon-intron junctions, and the 5' and 3' untranslated regions of WFS1. RESULTS: Nine different mutations in WFS1 (five of them novel) were identified in the nine patients. Six patients were homozygous for the following mutations: V412fs, S443R, W539X, V659fs. They developed diabetes at a mean age of 5·2 years. Three patients were compound-heterozygous for the following mutations: S167fs, Q392X, Y513fs, W648X, V779G. They developed diabetes at a mean age of 6·5 years. CONCLUSIONS: Mean age of diagnosis of diabetes among the Polish patients was typical for Wolfram syndrome; however, compound-heterozygous patients were slightly older at diabetes onset.

Donor-derived DNA in hair follicles of recipients after allogeneic hematopoietic stem cell transplantation.

The hair follicles of recipients of allogeneic hematopoietic SCT (HSCT) constitute the tissue with the greatest need for regeneration after high-dose chemotherapy. Previous studies have shown a lack of donor-derived DNA in the hair follicles of recipients. Therefore, we carried out a study to determine whether male donor-derived genetic material can be found in female recipients' hair follicles after HSCT. Fluorescent-based PCR with analyses of Y-chromosome STR (Y-STR) and RQ-PCR with the sex-determining region Y (SRY) were used independently to evaluate chimerism status. Our results proved the existence of donor-derived stem DNA in the recipients' hair follicle cells. This report undermines the validity of data indicating that hair follicle cells maintain 100% of recipient origin.

Population database on 15 autosomal STR loci in 1000 unrelated individuals from the Lodz region of Poland.

Allele frequency data and forensic efficiency parameters for 15 STR loci: D8S1179, D21S11, D7S820, CSF1PO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, vWA, TPOX, D18S51, D5S818, and FGA, were estimated from the sample of 1000 unrelated individuals from the Lodz region of Poland. The combined MP and PE for all 15 loci are 4.8 x 10(-18) and 0.9999989, respectively. The comparison of our data with other Polish populations revealed statistically significant differences in 6 out of 15 loci between Lodz and the Podlasie region of Poland.

Sonographic assessment of splanchnic arteries and the bowel wall.

The intestinal wall can be visualized using high resolution transabdominal ultrasound. The normal intestinal wall thickness in the terminal ileum, cecum, and right and left colon is <2mm when examined with graded compression. It is important to appreciate that a contracted intestinal segment can be misinterpreted as a thickened wall. Vascularisation can be mainly displayed in the second hyperechoic layer (submucosal layer) as well as vessels penetrating the muscularis propria. Imaging of the gastrointestinal wall is dependent on the experience of the examiner as well dependent on the equipment used. Acute or chronic inflammation of the intestinal wall is accompanied by increased perfusion of the mesentery, which can be displayed non-quantitatively with colour duplex. In contrast, ischemia is characterised by hypoperfusion of the mesenteric arteries and the bowel wall. The most promising sonographic approach in assessing splanchnic arteries and the bowel wall is combining the analysis of superior and inferior mesenteric inflow by pulsed Doppler scanning (systolic and diastolic velocities, resistance index) with the end-organ vascularity by colour Doppler imaging diminishing the influence of examination technique only displaying bowel wall vascularity. Colour Doppler imaging has been described as helpful in a variety of gastrointestinal disorders, particularly in patients with Crohn's disease, celiac disease, mesenteric artery stenosis and other ischemic gastrointestinal diseases, graft versus host disease and hemorrhagic segmental colitis.