Whole genome amplification of degraded and nondegraded DNA for forensic purposes

Degraded DNA is often analyzed in forensic genetics laboratories. Reliable analysis of degraded DNA is of great importance, since its results impact the quality and reliability of expert testimonies. Recently, a number of whole genome amplification (WGA) methods have been proposed as preamplification tools. They work on the premise of being able to generate microgram quantities of DNA from as little as the quantity of DNA from a single cell. We chose, investigated, and compared seven WGA methods to evaluate their ability to “recover” degraded and nondegraded DNA: degenerate oligonucleotide-primed PCR, primer extension preamplification PCR, GenomePlex™ WGA commercial kit (Sigma), multiple displacement amplification, GenomiPhi™ Amplification kit (Amersham Biosciences), restriction and circularization-aided rolling circle amplification, and blunt-end ligation-mediated WGA. The efficiency and reliability of those methods were analyzed and compared using SGMPlus, YFiler, mtDNA, and Y-chromosome SNP typing. The best results for nondegraded DNA were obtained with GenomiPhi and PEP methods. In the case of degraded DNA (200 bp), the best results were obtained with GenomePlex which successfully amplified also severely degraded DNA (100 bp), thus enabling correct typing of mtDNA and Y-SNP loci. WGA may be very useful in analysis of low copy number DNA or degraded DNA in forensic genetics, especially after introduction of some improvements (sample pooling and replicate DNA typing). ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00414-012-0764-9) contains supplementary material, which is available to authorized users

Wpływ polimorfizmów genów ACE i angiotensynogenu na rozwój otyłości brzusznej oraz strukturę i funkcję mięśnia lewej komory — wyniki 10-letniej prospektywnej obserwacji młodych zdrowych mężczyzn

Background Genetic and environmental factors play a major role in the pathogenesis of hypertension. The aim of this study was to investigate whether the insertion/deletion (I/D) polymorphism of angiotensin-converting enzyme (ACE) gene and angiotensinogen (AGT) gene increase the susceptibility to abdominal obesity and affect left ventricular function and structure in a 10-year follow-up of a young healthy male population Material and methods The study was carried out in a group of 68 normotensive subjects, aged 23 &#177; 3.5 years, mean &#177; SD. The subjects underwent the following procedures: anthropometric measurements, basic biochemical tests, office blood pressure measurements, ambulatory blood pressure measurements (ABPM), echocardiography. The I/D polymorphism within intron 16 of the ACE gene and AGT gene polymorphism (the allele M235T and T174M) were genotyped by polymerase chain reaction. Results No relationship between waist circumference and ACE, AGT polymorphisms was observed. The structural changes of the heart differed significantly in T174M AGT TT genotype with increased septum/ /posterior wall ratio compared to TM and MM: (1.20 &#177; 0.12 v. 1.1 &#177; 0.11 cm; p < 0.05). The II genotype of ACE gene was associated with an elevated A wave peak velocity in transmitral flow (0.56 &#177; 0.17 v. 0.48 &#177; 0.08 m/s; p < 0.05). Blood pressure was not related to the ACE and AGT polymorphisms. Conclusions In a healthy population, the ACE and AGT polymorphisms might contribute to the structural and functional changes of the heart, but not to development of abdominal obesity or hypertension. Arterial Hypertension 2011, vol. 15, no 4, pages 242&#8211;250.Wstęp W patogenezie nadciśnienia tętniczego istotną rolę odgrywa interakcja czynników genetycznych i środowiskowych. Celem niniejszego badania była ocena wpływu polimorfizmów genu enzymu konwertującego angiotensynę I (ACE) i angiotensynogenu (AGT) na rozwój otyłości brzusznej oraz strukturę i funkcję mięśnia lewej komory na podstawie 10-letniej prospektywnej obserwacji grupy młodych zdrowych mężczyzn. Materiał i metody Obserwacją objęto 68 studentów medycyny w wieku 23 &#177; 3,5 roku z prawidłowym ciśnieniem tętniczym. U wszystkich mężczyzn wykonano pomiar wskaźników antropometrycznych, podstawowe badania biochemiczne, pomiar ciśnienia tętniczego gabinetowy, całodobową automatyczną rejestrację ciśnienia tętniczego (ABPM) oraz badanie echokardiograficzne. Badanie polimorfizmu insercyjno-delecyjnego (I/D) w 16. intronie genu ACE oraz genu angiotensynogenu (polimorfizm T174M i M235T) przeprowadzono metodą łańcuchowej reakcji polimerazy (PCR). Wyniki W 10-letniej obserwacji zmiana obwodu talii nie miała związku z polimorfizmami genów ACE i AGT. W obrębie polimorfizmu T174M AGT, stosunek końcowo-rozkurczowego wymiaru grubości przegrody międzykomorowej do ściany tylnej był wyższy u homozygot TT w porównaniu z genotypem TM i MM (1,2 &#177; 0,12 v. 1,1 &#177; 0,11 cm; p < 0,05). Maksymalna prędkość czynnej fazy napełniania lewej komory serca (fala A) była istotnie wyższa w genotypie II ACE w porównaniu z resztą populacji (0,56 &#177; 0,17 v. 0,48 &#177; 0,08 m/s; p < 0,05). Nie znaleziono związku pomiędzy wysokością ciśnienia tętniczego a polimorfizmem T174M genu AGT i I/D genu ACE. Wnioski Polimorfizmy I/D genu ACE i T174M genu AGT determinują zmiany morfologii i funkcji mięśnia sercowego w populacji młodych zdrowych mężczyzn, ale nie mają związku z rozwojem otyłości brzusznej i nadciśnienia tętniczego w tej grupie osób. Nadciśnienie Tętnicze 2011, tom 15, nr 4, strony 242&#8211;250

A Unified Conformal Model for Fundamental Interactions without Dynamical Higgs Field

A Higgsless model for strong, electro-weak and gravitational interactions is proposed. This model is based on the local symmetry group SU(3)xSU(2)xU(1)xC where C is the local conformal symmetry group. The natural minimal conformally invariant form of total lagrangian is postulated. It contains all Standard Model fields and gravitational interaction. Using the unitary gauge and the conformal scale fixing conditions we can eliminate all four real components of the Higgs doublet in this model. However the masses of vector mesons, leptons and quarks are automatically generated and are given by the same formulas as in the conventional Standard Model. The gravitational sector is analyzed and it is shown that the model admits in the classical limit the Einsteinian form of gravitational interactions. No figures.Comment: 25 pages, preprin

Comparison of three methods of DNA extraction from human bones with different degrees of degradation

There is a necessity for deceased identification as a result of many accidents and sometimes bones are the only accessible source of DNA. So far, a universal method that allows for extraction of DNA from materials at different stages of degradation does not exist. The aims of this study were: the comparison of three methods of DNA extraction from bones with different degree of degradation and an evaluation of the usefulness of these methods in forensic genetics. The efficiency of DNA extraction, the degree of extract contamination by polymerase chain reaction (PCR) inhibitors and the possibility of determining the STR loci profile were especially being compared. Nuclear DNA from bones at different states of degradation was isolated using three methods: classical, organic phenol–chloroform extraction, DNA extraction from crystal aggregates and extraction by total demineralisation. Total demineralisation is the best method for most cases of DNA extraction from bones, although it does not provide pure DNA. DNA extraction from aggregates removes inhibitors much better and is also a good method of choice when identity determination of exhumed remains is necessary. In the case of not buried bones (remains found outside) total demineralisation or phenol–chloroform protocols are more efficient for successful DNA extraction

Population genetics of the amplified fragment length polymorphism and short tandem repeat type systems in the population of northern Poland (Gdańsk area)

This paper presents results of the population studies of the VNTR polymorphic systems of human DNA. DNA samples were taken from at least 106 unrelated persons living in the Gdansk area. DNA samples were subjected to the PCR amplification and PCR products separated on polyacrylamide gels. Allelic frequencies ofthree AMPFLPs systems and six STRs were calculated and homogeneity of alleles distribution between Gdansk and other population samples were compared. The presented data are the core of the planned population-based DNA indentification system for Poland

Directions of vision methods application in the works to date Łukasiewicz Research Network - Industrial Institute of Agricultural Engineering. Part 2.

W artykule przedstawiono wyniki prac związanych z rozpoznawaniem i analizą obrazu na potrzeby oceny zdrowotności upraw polowych, oceny jakości i równomierności siewu oraz automatyzacji ruchu pojazdów autonomicznych. Opisano algorytmy stosowane do oceny obrazów z użyciem sztucznej inteligencji oraz zastosowaną aparaturę.The article presents the results of work related to image analysis for the purposes of assessing the health of field crops, assessing the quality and uniformity of sowing, and automating the movement of autonomous vehicles. The algorithms used to evaluate images using artificial intelligence are described. The apparatus used was also presented

Directions of vision methods application in the works to date Łukasiewicz Research Network - Industrial Institute of Agricultural Engineering. Part 1.

W artykule przedstawiono wyniki prac związanych z analizą obrazu na potrzeby oceny zdrowotności upraw polowych, oceny jakości i równomierności siewu oraz automatyzacji ruchu pojazdów autonomicznych. Opisano algorytmy stosowane do oceny obrazów z użyciem sztucznej inteligencji oraz zastosowaną aparaturę.The article presents the results of work related to image analysis for the purposes of assessing the health of field crops, assessing the quality and uniformity of sowing, and automating the movement of autonomous vehicles. The algorithms used to evaluate images using artificial intelligence are described. The apparatus used was also presented

Strength and Weaknesses of Higher Education in Poland

W artykule przedstawiono udokumentowaną opinię o stanie systemu edukacji w Polsce. Oceniono pozycję Polski na tle innych krajów w oparciu o analizę indeksów narodowego potencjału intelektualnego. Wykazano stan niedofinansowania systemu kształcenia i skutki chronicznego niedofinansowania obszaru nauki. Pokazano ocenę stanu kadry nauczycieli akademickich, wpływ przeciążenia obowiązkami akademickimi na jakość kształcenia, brak jakiejkolwiek polityki rekrutacyjnej. W podsumowaniu zwrócono uwagę na konieczność podjęcia działań naprawczych.This paper presents an opinion on the state of the education system in Poland, which is based on the analysis of diverse indexes of national intellectual potential. It is shown that the education system is not financed enough and the area of science suffers from the chronic effects of underfunding. The paper presents an assessment of the state of academic staff, of the influence of an overload with academic responsibilities on the quality of education and of the lack of any recruitment policy. The summary emphasises the need to take corrective actions