[show abstract][hide abstract] ABSTRACT: The 2011 collaborative exercise of the ISFG Italian Working Group GeFI was aimed at validating the five ENFSI/EDNAP miniSTR loci D1S1656, D2S441, D10S1248, D12S391 and D22S1045. The protocol required to type at least 50 multilocus profiles from locally resident individuals and two blind bloodstains in duplicate (i.e., using at least two different commercial kits), and to send the electropherograms to the Organizing Committee. Nineteen laboratories distributed across Italy participated, collecting a total of 960 samples. Full concordance was found for the five new miniSTRs as observed from the comparison of 13,150 alleles. The inspection of the electropherograms allowed the identification of a very limited number of mistypings in the miniSTR genotypes thus contributing to the establishment of an high quality Italian database of frequencies.
[show abstract][hide abstract] ABSTRACT: Genetic markers represent a very important tool in forensic identification caseworks, in family relationships as well as in criminal analysis. The discrimination power of current genetic polymorphisms is so high that the inferential process can be efficiently used even in cases where direct knowledge on the genetic data of one of the terms in comparison is lacking. However in some cases despite the use of Probabilistic Expert Systems (FINEX) it is not always possible to achieve an acceptable percentage of paternity probability. Certainly one of these cases is the request to verify the relationship between two half-siblings of different gender in the absence of data from parents. In these cases it is not possible to use important tools such as polymorphisms of sexual chromosomes, so that the only possible approach is to increase the number of autosomal STRs. Therefore the authors decided to investigate 36 pairs of half-siblings with known relationships from different parts of Italy, using a high number of autosomal STRs. The aim of this study is to verify whether, increasing the number of autosomal STRs analyzed, the application of PES allows to achieve an acceptable value of paternity probability without availability of parents’ profiles.
Forensic Science International Genetics Supplement Series 03/2012;
[show abstract][hide abstract] ABSTRACT: We collected published and unpublished data from 17 contributing groups participating in the GeFI (Italian Forensic Geneticists). The total number of typed subjects was 1114 males and 777 females, coming from 11 regions of North, Centre, and South Italy, and Sardinia. Individual's multilocus genotypes included 4–12 loci. The total number of typed markers was 29, scattered along the X-chromosome genetic map in several clusters; the most used marker was DXS7423 (2429 gene copies); the mean number of subjects typed per marker was 336 for males and 208 for females. Data are available online.
Forensic Science International Genetics Supplement Series 12/2011;
[show abstract][hide abstract] ABSTRACT: The X-chromosomal markers are increasingly used in forensic genetics, particularly for relationship testing. Their use has become a valuable tool in complex cases of kinship but rather in criminal caseworks is still quite rare. In this paper the authors present a case of sexual assault in which the use of X chromosome polymorphisms has been crucial. The victim was a young immigrant woman found dead in her home. The main suspect was her husband. However, the couple lived in a community in which the particular cultural context suggested the involvement of the other males of the husband family. The suspect lived together with his brother, his father and his uncle. Generally, a useful tool to solve cases of sexual violence is undoubtedly the use of Y chromosome, but in this case this device could not discriminate between the four males involved. An additional factor has further complicated the situation: the most important biological evidence (typed with AmpFlSTR Identifiler and AmpFlSTR NGM) showed a mixed profile in which was very difficult to discriminate the suspects profiles. To solve the casework, the authors typed the victim, the suspects and the biological traces with 6 X-STRs in an homemade esaplex. The results showed the presence of victim and her husband profiles in the biological trace excluding his brother, father and uncle's profiles.
Forensic Science International Genetics Supplement Series 01/2011;
[show abstract][hide abstract] ABSTRACT: In recent years, European working groups (ENFSI-EDNAP) have strongly encouraged the development of new amplification kits that allow to obtain DNA profiles even on difficult samples (degraded samples, presence of inhibitory substances, LCN, mixtures, etc.). To enable the amplification of difficult biological samples the new loci were designed to achieve greater resistance to inhibitors and most robust and uniform amplification. With the aim of evaluating the performance of latest generation kits in real forensic caseworks the authors retested some bone samples previously analyzed in their laboratory over the past five years with the old commercial kits. The authors compared the performance of AmpFℓSTR® NGM™ PCR Amplification kit (Applied Biosystems) with PowerPlex® ESX 17 systems (Promega). The twelve analyzed samples came from eight exhumed corpses and four bodies found outdoor in the advanced stage of putrefaction. For the genetic investigation the authors used a piece of bone taken from the femur. In conclusion, the authors can claim that the latest generation kits have proved to be decisive in all cases, including those where the previous use of traditional kits did not produce reliable and uniquely interpretable results.
Forensic Science International Genetics Supplement Series 01/2011;
[show abstract][hide abstract] ABSTRACT: Mitochondrial DNA (mtDNA) U/K and J/T are sister haplogroups within the superhaplogroup R. They are both common in Europe, with a combined overall frequency similar to the one reported for H, the most common European haplogroup (40-50%). In this study, we selected 159 Italian subjects, already ascribed to U/K and J/T by RFLP typing, and assigned each mtDNA to specific clades/subclades by investigating at least one diagnostic coding region SNP. For each sister haplogroup, one multiplex PCR and one SNaPshot minisequencing reaction were set up targeting 16 U/K and 7 J/T coding region SNPs. Each mtDNA sample was clearly assigned to a specific subclade, which could be further subdivided into several minor sub-branches according to peculiar HVS I/II motifs. Such a molecular dissection of haplogroups U/K and J/T could be extremely useful to reduce the overall analysis time and labor intensive sequencing procedures in high volume forensic casework, for example when it is important to rapidly exclude samples in order to restrict the number of suspects.
[show abstract][hide abstract] ABSTRACT: A collaborative study was carried out by the Italian ISFG Working Group in order to improve the data on Y-STR mutations at the loci mostly used in forensic analysis, following recommendations of the ISFG DNA Commission.
The knowledge on Y-STR mutation rates needs to be considered in the paternity probabilities, especially in deficiency cases of disputed paternity involving male offspring where the alleged father is not available for DNA analysis. Furthermore, the mutation rate represents a precious tool to estimate the local and temporal origin of a given Y-SNP based haplogroup.
The sample consisted of 433 father/son pairs from paternity cases in 15 different laboratories from Italy. The biological relationship of all father/son pairs was previously confirmed by using autosomal microsatellites. The laboratories used AmpFlSTR YFiler kit (AB) and PowerPlex Y System (Promega); DYS19, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS385, DYS437, DYS438, DYS439, DYS448, DYS456, DYS458, GATA C4, and GATA H4.1 data were collected. The participants were also asked to provide the age of the biological father and, if possible, male descendants beyond the first generation.
20 mutations were observed among all of the allele transfers in the sample (19 single step and 1 double step), and mutations in the same father/son pair were found in three cases. Locus-specific mutation rates were calculated. Forensic implication of the average age of the father as well as the number of locus deletions and amplifications were discussed.
[show abstract][hide abstract] ABSTRACT: A case of a woman killed in Perugia is reported. The woman was beaten to death and the body showed evidence of bites, kicks and punches. The request of the Court was to verify the presence of bites and if they belonged to humans. Morphological examination and genetic analysis with human Y-specific markers were performed in order to verify the origin of the bites. The DNA profile from the surrounded area of the traces was compared with the profile of the victim's husband (the suspect).The results showed a match between the profile of the suspect and that of the traces for all loci examined. Due to the fact that also other relatives of the husband's male lineage lived in the same house, it was not possible to identify the man who really contributed to the traces. Therefore, the analysis was implemented with autosomal STR markers, which showed a mixed genetic profile. In order to verify the number and the identity of the contributors, statistical analysis based upon peak area information was performed with Probabilistic Expert Systems.
Forensic Science International Genetics Supplement Series 09/2009;
[show abstract][hide abstract] ABSTRACT: Forensic identification with genetic markers is currently experiencing various scenarios of inference, spanning from direct to indirect comparisons in matter of family relationship and criminal analysis. The discrimination power of current genetic polymorphisms is such that the inferential process underlying this field of human identification can efficiently be deployed even in cases where there is a lack of direct knowledge on the genetic asset on one of the terms in comparison. The most promising of these software facilities are the probabilistic expert systems (PES). To verify the power of these systems to confirm relationships among individuals in defective cases, we analyzed four different groups of complex families with known relationships. To test the goodness of the software employed (FINEX), we created different pedigrees by deleting one or more individuals from each family and calculating the different probability values obtained.
Forensic Science International Genetics Supplement Series 01/2009;
[show abstract][hide abstract] ABSTRACT: MiniSTR loci have demonstrated to be an effective tool to recover genetic information from degraded sample due to the small PCR products. The use of these non-CODIS miniSTRs can increase the probability that a degraded sample can be typed and in addition these systems can be used in complex paternity cases where more markers are needed.Six autosomal loci (D10S1248, D14S1434, D22S1045, D4S2364, D2S441, D1S1677) were investigated in 200 individuals from Italian regions, Umbria and Sardinia.Statistical analysis and comparison between two groups were performed.
Forensic Science International Genetics Supplement Series 01/2009; 2(1):367-368.
[show abstract][hide abstract] ABSTRACT: Eight X-chromosome STR were investigated in two groups of individuals from two Italian regions, Umbria and Sardinia. These two regions have a very different history. Umbria, situated in the centre of Italy and without defensive natural barriers, in the course of the centuries has been object of many invasions and appropriations by foreign people. On the contrary, Sardinia, an island in the centre of the Mediterranean sea, has endured a minor number of foreign invasions. The ChrX markers were amplified in a pentaplex (DXS6789, DXS7423, DXS6807, DXS101, DXS8377) and a triplex (DXS10074, DXS10075, DXS10079) that represents a haplotype. The DNA was extracted from 200 blood samples using the QIAmp DNA Minikit (Qiagen TM). Amplification products were detected on an ABI PRISM 310 Genetic Analyzer (Applied Biosystems), using the same dye labels, run conditions, standard (GeneScan 500 Liz) and matrix file of AmpFlSTRIdentifiler. Statistical analyses for all the loci and intergroup comparisons between the two regional groups of individuals were performed.
Forensic Science International Genetics Supplement Series 01/2009;
[show abstract][hide abstract] ABSTRACT: In the course of an investigation to identify a burns-related death victim a particular case of paternity test was introduced. In a locality near Spoleto (Umbria), an oil mill burned down causing the death of four men. One of them was identified comparing his genetic profile with those of his relatives: a daughter and a brother. The wife was not available for the analysis. Data obtained with Identifiler showed a genetic compatibility between the man and his daughter. By chance we observed that a genetic compatibility was also evident between the daughter and the other man analyzed, the uncle. To clarify this situation we implemented the panel of markers analyzed, adding PowerPlex 16, 6 miniSTRs (NC01 and NC02) and 5 STRs of X chromosome.
Forensic Science International Genetics Supplement Series 02/2008;
[show abstract][hide abstract] ABSTRACT: Within an Italian collaborative exercise on the extended haplotype of the Y-chromosome, 1288 subjects were typed by the AmpFlSTR YFiler Amplification Kit (AB Applied Biosystems) and other 526 were typed by the PowerPlex Y® System (Promega). The sampling scheme included either a “regional” or a “local” recruitment, the first referring to individuals born in the region of the participating lab, the second referring to individuals coming from small villages. Total sample sizes were N=954 and 860, respectively. A significant decrease of haplotype diversity was found in the local samples. The results may be of interest in forensic applications of the Y-chromosome.
Forensic Science International Genetics Supplement Series 01/2008; 1(1):239-241.
[show abstract][hide abstract] ABSTRACT: A sample of 1070 individuals was collected by 12 collaborating forensic laboratories from Central and Northern Italy, and typed by the so-called “NC01” triplex, including the three loci D10S1248, D14S1434, and D22S1045. Heterozygosities were 0.771, 0.717, and 0.720, respectively.
Forensic Science International Genetics Supplement Series 01/2008; 1(1):373-374.
[show abstract][hide abstract] ABSTRACT: L'uso di preparati istologici a fini identificativi nei casi di interesse giudiziario
sta assumendo i caratteri di una metodica ampiamente diffusa sia nel campo della
responsabilità medica sia per le indagini di parentela.
Il presente lavoro è finalizzato alla valutazione degli effetti degradativi tempo-correlati della formaldeide sul DNA nonché dell'instabilità dei microsatelliti a carico dei tessuti tumorali. È stata, inoltre, valutata l'azione della formaldeide su organi differenti. Modificazioni genetiche sono risultate presenti
in percentuali significative sia nei tessuti tumorali sia in tessuti sani sottoposti agli
usuali protocolli di allestimento di preparati istologici. L azione della formalina su
tessuti tumorali aumenta il numero e il grado delle alterazioni genetiche. Esiste una diretta correlazione tra tempo di pertinenza del campione ii formalina e gravità
del danno subito dalle strutture nucleari. Riguardo all'azione della formaldeide
su diversi organi esaminati non è emersa una variabilità significativa tra organi
dello stesso individuo.
[show abstract][hide abstract] ABSTRACT: T-cell homeostasis is regulated by several molecules; among these, interleukin (IL)-7 plays an essential role in the survival and homeostatic proliferation of peripheral naive T cells. In a previous study, we investigated whether human mesenchymal stromal cells (MSCs) could be engineered with the IL-7 gene to produce functional level of this cytokine. In the present study, we analyzed the impact of different quantities of IL-7 produced by MSCs on the survival and proliferation of a negative immunoselected naive (CD3(+)/CD45RA(+)) T-cell population. Co-cultivation of peripheral naive T cells with MSCs producing low (16 pg/mL) or high (1000 pg/mL) IL-7 levels or in the presence of exogenous IL-7 (0.01 ng/mL and 100 ng/mL) maintained the CD3(+)/CD45RA(+) naive T-cell phenotype. Chemokine receptor CCR7(+) expression was also maintained among this T-cell population. Naive T-cell molecular characteristics were maintained as assessed by the Vbeta spectratyping complexity score, which showed the maintenance of a broad T-cell repertoire. No Th1 or Th2 differentiation was observed, as assessed by interferon-gamma or IL-4 accumulation. In contrast, only MSCs producing high amounts of IL-7 caused increased activation (CD25 31.2% +/- 12% vs 10% +/- 3.5%; P < .05), proliferation (CD71 17.8+/-7% vs 9.3%+/-3, P < .05), apoptosis (assessed by annexin V: 18.6% +/- 5% vs 14.9% +/- 2.6%; P > .05), and the phase S cell cycle (15% vs 6.9%, P > .05). Exogenous IL-7 exhibited no significant effect. In conclusion, we demonstrated that IL-7 produced by MSCs has a dose-independent effect on naive T-cell survival while exerting a dose-dependent effect on activation/proliferation. Due to the continuous production of IL-7 by engineered cells, our system is more efficacious than exogenous IL-7.
Biology of Blood and Marrow Transplantation 01/2007; 12(12):1250-60. · 3.94 Impact Factor
[show abstract][hide abstract] ABSTRACT: Results from a collaborative exercise with proficiency testing conducted by 20 Italian laboratories on the 15 loci included in the Identifiler kit were analyzed by allele sharing methods and by standard population genetics tests. The validated database, including about 1500 subjects, was merged with that of a previous exercise conducted on nine loci, and the resulting allele frequencies, subdivided by Italian region, were published on-line.
Forensic Science International 02/2006; 156(2-3):266-8. · 2.31 Impact Factor