Development of an Italian RM Y-STR haplotype database: Results of the 2013 GEFI collaborative exercise.

Recently introduced rapidly mutating Y-chromosomal short tandem repeat (RM Y-STR) loci, displaying a multiple-fold higher mutation rate relative to any other Y-STRs, including those conventionally used in forensic casework, have been demonstrated to improve the resolution of male lineage differentiation and to allow male relative separation usually impossible with standard Y-STRs. However, large and geographically-detailed frequency haplotype databases are required to estimate the statistical weight of RM Y-STR haplotype matches if observed in forensic casework. With this in mind, the Italian Working Group (GEFI) of the International Society for Forensic Genetics launched a collaborative exercise aimed at generating an Italian quality controlled forensic RM Y-STR haplotype database. Overall 1509 male individuals from 13 regional populations covering northern, central and southern areas of the Italian peninsula plus Sicily were collected, including both "rural" and "urban" samples classified according to population density in the sampling area. A subset of individuals was additionally genotyped for Y-STR loci included in the Yfiler and PowerPlex Y23 (PPY23) systems (75% and 62%, respectively), allowing the comparison of RM and conventional Y-STRs. Considering the whole set of 13 RM Y-STRs, 1501 unique haplotypes were observed among the 1509 sampled Italian men with a haplotype diversity of 0.999996, largely superior to Yfiler and PPY23 with 0.999914 and 0.999950, respectively. AMOVA indicated that 99.996% of the haplotype variation was within populations, confirming that genetic-geographic structure is almost undetected by RM Y-STRs. Haplotype sharing among regional Italian populations was not observed at all with the complete set of 13 RM Y-STRs. Haplotype sharing within Italian populations was very rare (0.27% non-unique haplotypes), and lower in urban (0.22%) than rural (0.29%) areas. Additionally, 422 father-son pairs were investigated, and 20.1% of them could be discriminated by the whole set of 13 RM Y-STRs, which was very close to the theoretically expected estimate of 19.5% given the mutation rates of the markers used. Results obtained from a high-coverage Italian haplotype dataset confirm on the regional scale the exceptional ability of RM Y-STRs to resolve male lineages previously observed globally, and attest the unsurpassed value of RM Y-STRs for male-relative differentiation purposes.

Population data for 17 Y-chromosome STRs in a sample from Apulia (Southern Italy).

The 17 Y-STR loci included in the AmpFLSTR Yfiler PCR Amplification Kit were analyzed in 98 unrelated healthy males from Apulia (Southern Italy). A total of 97 different haplotypes were identified, of which 96 haplotypes were unique and 1 occurred twice. Allele frequencies for each Y-STR locus in pooled sample and estimated value of gene diversity (GD) were evaluated. The lowest value of GD was observed for DYS392 (0.126) and the highest one (0.936) for DYS385. The HD (haplotype diversity) for the studied Y-STR set showed a value of 0.9994, with an HMP (haplotype match probability) value of 0.0006, while the overall DC was 98.98%. Microvariant alleles were found for the DYS458 and DYS385 markers and sequenced. Furthermore, Φ(st)-based genetic distance computation and pair-wise analysis of molecular variance (AMOVA) test were carried out. When comparing our population with the Apulia sample previously investigated, the AMOVA analysis detected no evidence for significant differentiation. The comparison with all Italian populations submitted to the YHRD website showed no relevant differences with all Southern Italian populations (San Giorgio La Molara, Belvedere, Trapani and Catania) and significant genetic deviation with all Northern Italian populations (Udine, Biella, La Spezia, Modena, Ravenna, Marche and North Sardinia). Moreover, the other populations and meta-populations belonging to the whole Mediterranean area (Croatia, Macedonia, Albania, Greece, Turkey, Israel, Libya, Tunisia, Algeria, Morocco and Spain) were different from our Apulia sample. The data were submitted to YHRD.

Allele frequencies of the new European Standard Set (ESS) loci in a population of Apulia (Southern Italy).

Allele frequencies of five miniSTRs loci (D1S1656, D2S441, D12S391, D10S1248 and D22S1045) included in the new European Standard Set (ESS) were calculated from a sample of 150 unrelated individuals from Apulia, a Region of Southern Italy. Two different PCR Amplification Kits were used, in order to evaluate the concordance of the genotypes. The results obtained with the two kits showed no differences in all genotype profiles. No deviation from Hardy-Weinberg expectations was detected at either locus. Moreover genetic analysis using Fst estimation showed no evidence for differentiation at the five new loci between Apulia and Italian populations. The high levels of polymorphisms of the analyzed markers in the Apulian population allow to confirm that these markers are useful tools in paternity and forensic analysis from degraded DNA samples.

The 2011 GeFI collaborative exercise. Concordance study, proficiency testing and Italian population data on the new ENFSI/EDNAP loci D1S1656, D2S441, D10S1248, D12S391, D22S1045.

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.

Endoluminal repair of infrarenal abdominal aortic aneurysms using a modular stent-graft: one-year clinical results from a European multicentre trial.

This is a report of a prospective study to evaluate the safety and efficacy of the Medtronic AneuRx stent-graft. Patients with an infrarenal aneurysm with a proximal neck length of greater than 10 mm and a neck diameter not greater than 26 mm and iliac artery diameters of at least 6 mm were accepted for endovascular repair using the Medtronic AneuRx modular stent-graft. A total of 104 patients were included in the study. Transfemoral placement of the bifurcated stent-graft was successful in 102 of 104 patients. Two conversions were performed. The mean operating time was 148 min (range 75-480) and the mean blood loss was 605 ml (range 100-2900). The mean follow-up was 15 months (range 12-21). Complications were rare and no kinking or migration occurred. At follow-up, four endoleaks persisted after 12 months. The Medtronic AneuRx stent-graft is a safe and efficacious alternative to open abdominal aortic aneurysm repair with excellent early results.

[Transluminal treatment in aneurysm of the abdominal aorta. Role of diagnostic imaging]. / Trattamento transluminale degli aneurismi dell'aorta addominale. Ruolo della diagnostica per immagini.

PURPOSE: To emphasize the importance of diagnostic imaging: a) in the selection of patients to be treated with the transluminal approach b) during stent-graft positioning c) in the follow-up of treated patients. MATERIAL AND METHODS: From January 1997 to May 1998, twenty-five patients with abdominal aortic aneurysms (AAA) were treated with transfemoral stent-grafts (AneuRx Medtronic). All patients were submitted to a preoperative study including digital angiography (DSA) and Spiral CT. Intraoperatively they underwent DSA and intravascular ultrasound (IVUS). Follow-up was performed with Spiral CT. RESULTS: Twenty-four bifurcated and one straight device were inserted in twenty-five patients with AAA. In two cases it was impossible to position the endoprosthesis due to narrow or tortuous artery access. Following the intention to treat criteria, the technical success rate was 92.5%. All the stents were patent and no dislodgement was observed at follow-up. Partial thrombosis of the stent was observed in three patients. Owing to incomplete distal covering, early endoleak occurred in one patient with an aortoiliac aneurysm; the positioning of two cuffs allowed a successful outcome. CT examination performed 6 months after positioning revealed the presence of endoleaks in three patients, due to persistence of lumbar and inferior mesenteric artery patency. The AAA was no more appreciable in five of the ten patients submitted to CT follow-up one year after the procedure. In three of ten cases it was reduced in size and in two patients there was no change. DISCUSSION: Spiral CT plays a basic role in the selection of patients because it helps assess the length and diameter of the proximal neck, thus permitting to choose the device to be inserted. Preoperative DSA is mandatory in the evaluation of size and tortuosity of the iliac arteries. IVUS allows to monitor the site of delivery during the maneuver and to make the final measurements while DSA plays a role in checking the correct positioning of the device and excluding the presence of endoleaks at the end of the procedure. Late follow-up with Spiral CT aims at demonstrating possible malfunctioning of the endoprosthesis and confirms the definitive exclusion of AAA. CONCLUSIONS: Diagnostic imaging plays a basic role in the endovascular treatment of AAA, much more than that required for traditional surgical treatment. In particular, pretreatment planning is critical and requires sophisticated imaging including Spiral CT with 3D reconstruction and angiographic evaluation using catheters with calibrated markers.

Carotid plaque characterization using digital image processing and its potential in future studies of carotid endarterectomy and angioplasty.

PURPOSE: To corroborate the validity of a computerized methodology for evaluating carotid lesions at risk for stroke based on plaque echogenicity. METHODS: The records of 96 carotid endarterectomy patients (59 men; median age 69.5 years, range 52 to 83) with stenoses > 50% were studied retrospectively. Forty-one patients (43%) had been symptomatic preoperatively. All patients had undergone computed tomography (CT) to detect infarction in the carotid territory and a duplex scan to measure carotid stenosis. Plaque echogenicity was analyzed by computer, expressing the echodensity in terms of the gray scale median (GSM). The incidence of CT-documented cerebral infarction was analyzed in relation to symptomatology, percent stenosis, and echodensity. RESULTS: Symptoms correlated well with CT evidence of brain infarction: 32% of symptomatic patients had a positive CT scan versus 16% for asymptomatic plaques (p = 0.076). The mean GSM value was 56 +/- 14 for plaques associated with negative CT scans and 38 +/- 13 for plaques from patients with positive scans (p < 0.0001). However, there was no difference in the GSM value between plaques with > or < 70% stenosis. Furthermore, the incidence of CT infarction was 40% in the cerebral territory of carotid plaques with a GSM value < 50 and only 9% in those with a GSM > 50 (p < 0.001). CONCLUSIONS: Computerized analysis of plaque echogenicity appears to provide clinically useful data that correlates with the incidence of cerebral infarction and symptoms. This method of analyzing plaque echolucency could be used as a screening tool for carotid stent studies to identify high-risk lesions better suited to conventional surgical treatment.

European multicentre experience with modular device (Medtronic Aneurx) for the endoluminal repair of infrarenal abdominal aortic aneurysms.

OBJECTIVE: Transfemoral endoluminal repair of AAA, introduced for the first time in the early 90's, has become a very promising alternative to conventional open repair and more and more centers are reporting satisfactory postoperative results in a high percentage of cases. Straight and bifurcated grafts represent the devices available on the market at present and aortic, as well as iliac aneurysmal lesions can be safely treated through a transfemoral approach. The possibility to indicate an endovascular AAA repair is related to the configuration (length and size) of the proximal and distal necks, tortuosity and calcification of the access arteries and to vascular and non-vascular comorbidities, which afflict the patients. The objective of our study was to evaluate the early and late postoperative results in a series of patients affected by infrarenal AAA, who underwent endoluminal repair. MATERIALS AND METHODS: From December 1996 to 31 October 1997 in 5 different European Centers, 100 Medtronic AneuRx bifurcated stent grafts were implanted for infrarenal abdominal aortic aneurysms. The diameter of the AAA varied from 33 to 77 mm (average 64 mm) and the mean age of the patients was 70.8 years (51-87 years). In one patient with a 33 mm diameter of the aneurysm, the surgical procedure was indicated because the size of the aneurysm had increased by 5 mm, compared to the previous control made 2 months before. In addition the aneurysm became symptomatic. There were 92 male and 8 female patients. The average time of the surgical procedure was 150 minutes (75-480 minutes) with an average blood loss of 570 ml (100-2,600 ml).

New diagnostic and screening possibilities in the stenobliterative lesions of the deep visceral vessels.

The evolution and continuous improvement of the ultrasonic diagnostic devices allows an hemodynamic and morphological exploration of the deep abdominal vessels that until now seemed to be precluded to non invasive diagnostic techniques. For example, the angiodynography today advantageously substitutes classical urographic investigation during the initial screening of the hypertensive patients suspected of stenotic lesions of the renal arteries. In fact the ecocolordopplersonography allows not only a morphological evaluation of the parenchyma and of the renal vascular tree, but also supplies a quantitative measurement of the blood flow through the renal artery. Still more important are the possibilities offered by these devices in the study of the acute or chronic intestinal ischemia, in which until now a sure diagnosis was only possible using invasive techniques. Performing a study of the mesenteric vascularization urgently, sometimes is possible a decisive demonstration of an ischemic pathology where the clinical abdominal picture provides only uncertain diagnostic elements.

[Subrenal aortic occlusion]. / L'obliterazione aortica sottorenale.

The juxtarenal aortic occlusion is an infrequent but not exceptional condition determining several problems during the corrective procedure. The incidence of this pathological picture is around 10% of the patients affected by steno-obliterative lesions of the subrenal aorta. The danger of this morbid condition derives mainly from the risks threatening the renal function which sometimes is already endangered before surgical intervention. The operation however implies high additional risks because some surgical procedures require suprarenal aortic clamping and because during the surgical manoeuvres embolization could occur in the lumen of renal arteries. This study analyzes 206 patients who underwent surgical intervention for subrenal aortic occlusion during the last 15 years. 11 patients were operated on in emergency because of lower limbs ischaemia. 4 patients presented a complete thrombosis of an aneurysmatic lesion of the subrenal aorta. The study is aimed at analyzing the renal behaviour in the postoperative period, comparing its functionality with the preoperative parameters. A second aspect which will be considered refers to the type of surgical procedure.