Applicability of 'Toolkit for Safety Assessment' tool to interventional radiology using probabilistic risk assessment techniques.

Interventional radiology brings extensive benefits to patients. Nevertheless, certain procedures may result in high doses of radiation, leading to health risks to occupationally exposed individuals (OEIs). Therefore, a more comprehensive risk analysis is essential to ensuring safety and minimising radiation exposures for all OEIs. The Toolkit for Safety Assessment (TOKSA) tool performs risk assessments based on the concepts described in 'General Safety Requirements' Part 3 (Radiation Protection and Safety of Radiation Sources: International Basic Safety Standards) and Part 4 (Safety Assessment for Facilities and Activities). This tool was developed based on the 'Ibero-American Forum of Radiological and Nuclear Regulatory Agencies' risk models and can promote the use of the risk assessment processes by OEIs. The aim of this study was to experimentally analyse the applicability of the TOKSA tool in interventional radiology with the use/support of probabilistic risk assessment techniques. The results were used to reduce the risks associated with a hemodynamics room in a hospital in Belo Horizonte, Brazil.

The miR-139-5p regulates proliferation of supratentorial paediatric low-grade gliomas by targeting the PI3K/AKT/mTORC1 signalling.

AIMS: Paediatric low-grade gliomas (pLGGs) are a heterogeneous group of brain tumours associated with a high overall survival: however, they are prone to recur and supratentorial lesions are difficult to resect, being associated with high percentage of disease recurrence. Our aim was to shed light on the biology of pLGGs. METHODS: We performed microRNA profiling on 45 fresh-frozen grade I tumour samples of various histological classes, resected from patients aged ≤16 years. We identified 93 microRNAs specifically dysregulated in tumours as compared to non-neoplastic brain tissue. Pathway analysis of the microRNAs signature revealed PI3K/AKT signalling as one of the centrally enriched oncogenic signalling. To date, activation of the PI3K/AKT pathway in pLGGs has been reported, although activation mechanisms have not been fully investigated yet. RESULTS: One of the most markedly down-regulated microRNAs in our supratentorial pLGGs cohort was miR-139-5p, whose targets include the gene encoding the PI3K's (phosphatidylinositol 3-kinase) catalytic unit, PIK3CA. We investigated the role of miR-139-5p in regulating PI3K/AKT signalling by the use of human cell cultures derived from supratentorial pLGGs. MiR-139-5p overexpression inhibited pLGG cell proliferation and decreased the phosphorylation of PI3K target AKT and phosphorylated-p70 S6 kinase (p-p70 S6K), a hallmark of PI3K/AKT/mTORC1 signalling activation. The effect of miR-139-5p was mediated by PI3K inhibition, as suggested by the decrease in proliferation and phosphorylation of AKT and p70 S6K after treatment with the direct PI3K inhibitor LY294002. CONCLUSIONS: These findings provide the first evidence that down-regulation of miR-139-5p in supratentorial pLGG drives cell proliferation by derepressing PI3K/AKT signalling.

Expanding the mutational spectrum associated to neural tube defects: literature revision and description of novel VANGL1 mutations.

BACKGROUND: Neural Tube Defects (NTD) are a common class of birth defects that occur in approximately 1 in 1000 live births. Both genetic and nongenetic factors are involved in the etiology of NTD. Planar cell polarity (PCP) genes plays a critical role in neural tube closure in model organisms. Studies in humans have identified nonsynonymous mutations in PCP pathway genes, including the VANGL genes, that may play a role as risk factors for NTD. METHODS: Here, we present the results of VANGL1 and VANGL2 mutational screening in a series of 53 NTD patients and 27 couples with a previous NTD affected pregnancy. RESULTS: We identified three heterozygous missense variants in VANGL1, p.Ala187Val, p.Asp389His, and p.Arg517His, that are absent in controls and predicted to be detrimental on the protein function and, thus, we expanded the mutational spectrum of VANGL1 in NTD cases. We did not identify any new variants having an evident pathogenic effect on protein function in VANGL2. Moreover, we reviewed all the rare nonsynonymous or synonymous variants of VANGL1 and VANGL2 found in patients and controls so far published and re-evaluated them for their pathogenic role by in silico prediction tools. Association tests were performed to demonstrate the enrichment of deleterious variants in reviewed cases versus controls from Exome Variant Server (EVS). CONCLUSION: We showed a significant (p = 7.0E-5) association between VANGL1 rare genetic variants, especially missense mutations, and NTDs risk.

Successful isolation and long-term establishment of a cell line with stem cell-like features from an anaplastic medulloblastoma.

AIMS: Herein we report on the successful isolation and establishment of a novel, long-term, primary, neurosphere-like cell line called 1603-MED from a 5-year-old boy affected by a highly aggressive anaplastic medulloblastoma. METHODS: Elaboration of the new protocol for neurosphere assay is extensively discussed, together with a complete immuno-histochemical and cytogenetic characterization of 1603-MED. RESULTS: Clinical course and histopathology are briefly discussed. The 1603-MED possesses a high capacity for proliferation, CD133 expression, self-renewal and differentiation, thus indicating that anaplastic medulloblastoma contains a subpopulation of cancer stem cells as observed in classic medulloblastoma. CONCLUSIONS: 1603-MED provides us with the first in vitro model of anaplastic medulloblastoma that may be suitable for studying both tumour progression and the genetic mechanisms related to therapy resistance, and may lead to the development and testing of chemosensitivity and new therapeutic targets.

Synthesis and mass spectroscopy kinetics of a novel ternary copper(II) complex with cytotoxic activity against cancer cells.

The X-ray structure of the [Cu(I-hip)(phen)2](+).(I-hip-).(H2O)7 complex (1) (where I-hipH is referred to o-iodohippuric acid and phen is 1,10-phenanthroline) and its binary synthetic intermediate [Cu(I-hip)2(H2O)3].(H2O)2 (2) have been solved and characterized by different techniques. This ternary [Cu(I-hip)(phen)2]+.(I-hip-).7H2O complex generates the copper(I) complex [Cu(phen)2]+ in aqueous solution without the addition of any external reductant, possibly by an intramolecular red-ox process in the presence of oxygen; the ESI-HRMS spectra (electrospray ionization-high resolution mass spectroscopy) detect these species and 24h after the solution, [Cu(phen)2]+ is the main product. The complex 1 is capable of cleaving DNA. To evaluate the biological properties, we carried out: cell culture, cell proliferation assays, cell cycle analysis, and electrophoresis (SDS-PAGE) and immunoblotting. Complex 1 induced apoptosis of A549 cells at low nanomolar and induced marked decreases of cancer cells at concentrations that did not change adipocyte survival. These data indicate that the parent complex is a potential anticancer drug.

Whole-genome sequencing as standard practice for the analysis of clonality in outbreaks of meticillin-resistant Staphylococcus aureus in a paediatric setting.

Meticillin-resistant Staphylococcus aureus (MRSA) is one of the leading causes of hospital-associated infections. This study investigated the potential use of whole-genome sequencing (WGS) for surveillance purposes by re-examining MRSA strains related to past outbreaks among hospitalized paediatric patients. WGS data ameliorated the genotypic profile previously obtained with Sanger sequencing and pulsed-field gel electrophoresis typing, and discriminated between strains that were related and unrelated to the outbreaks. This allowed strain clonality to be defined with a higher level of resolution than achieved previously. This study demonstrates the potential of WGS to trace hospital outbreaks, which may lead to WGS becoming standard practice in outbreak investigations.

Effect of metal complexes of acyclovir and its acetylated derivative on Herpes simplex virus 1 and Herpes simplex virus 2 replication.

The effect of zinc, nickel, cobalt and cadmium complexes of acyclovir (ACV) and its omicron-acetylated derivative (Ac-ACV) on the replication of wild type (wt) and ACV-resistant (ACV(R)) strains of Herpes simplex virus 1 (HSV-1) and Herpes simplex virus 2 (HSV-2) was examined. According to cytotoxicity, these compounds followed the order Ni-ACV chloride > Cd-ACV 3 Ni-ACV nitrate > ACV = Zn-ACV nitrate = Ac-ACV = Zn-Ac-ACV > Zn-ACV chloride > Co-ACV. Besides Ac-ACV, the only active complexes in inhibiting virus replication were Zn-ACV nitrate and Zn-Ac-ACV, which effectively suppressed the growth of both wt and ACVR strains of HSV-1 and HSV-2. The most active and most selective inhibitor of the growth of ACVR strains of HSV-1 and HSV-2 was Ac-ACV; its EC50 and SI were 100 and 10 times higher than those of ACV, respectively. Zn-Ac-ACV was less active than Ac-ACV, obviously due to the stability of the complex. Zn-ACV nitrate was active against both wt and ACVR strains of HSV-1; its activity and selectivity were 100 and 75 times higher than those of ACV, respectively. Ac-ACV and Zn-Ac-ACV suppressed the pre-mitotic arrest caused by HSV-1 infection during the first 2 hrs of infection and later on restored the cell division.

An improved method for HLA-B and -C supratyping.

A growing body of evidence links the analysis of the KIR genotype and the presence of their HLA-B and -C ligands to a wide repertoire of human diseases. We noticed that, using a panel of 184 Caucasoid donors, a limited number of HLA alleles were incorrectly supratyped by previously described pyrosequence-based assays. Here we describe a simple implementation of the reported methods that corrects all the discrepancies found with HLA-B and -C molecular typing and allows establishing a quick and high-throughput method for the determination of HLA-Bw4 I(80), Bw4T(80), Bw6 and HLA-C1 or -C2 supratype.

DNA fingerprinting of yeast strains by restriction enzyme analysis.

Restriction endonuclease analysis was used as a new method to obtain genomic DNA fingerprints in yeast. Fifteen yeast strains belonging to the genera Saccharomyces and Zygosaccharomyces were examined. Restriction fragments obtained with ApaI or KspI endonucleases were separated by SDS-PAGE and silver-stained. Analysis of the fingerprints showed that restriction enzyme digestion of genomic DNA can be successfully applied to yeast, enabling the differentiation between strains belonging to different or to the same species or genera.

Ternary complexes metal [Co(II), Ni(II), Cu(II) and Zn(II)]--ortho-iodohippurate (I-hip)--acyclovir. X-ray characterization of isostructural [(Co, Ni or Zn)(I-hip)(2)(ACV)(H(2)O)(3)] with stacking as a recognition factor.

Four ternary metal--ortho-iodohippurate (I-hip)--acyclovir (ACV) complexes, [M(I-hip)(2)(ACV)(H(2)O)(3)] where M is Co(II) (1), Ni(II) (2), Cu (3) and Zn(II) have been obtained by reaction between the corresponding binary complexes M(II)(I-hip)(2)xnH(2)O and ACV. Three ternary complexes (M=Co, Ni and Zn) and the corresponding Zn(II)--ortho-iodohippurate binary derivative have been structurally characterized by X-ray diffraction: The studies show these three ternary complexes are isostructural and present, in solid state, an interesting stacking between the nucleobase and the aryl ring of the hippurate moiety, which probably promotes the formation of ternary complexes. Moreover, the two different ligands interact between them by means of ancillary hydrogen bonds with water molecules coordinated to the metal ion. It must be mentioned that these two recognition factors, hydrogen bonds plus stacking, could explain the reason for the isostructurality of these ternary derivatives with so different three metal ions, with diverses trends in coordination numbers and geometries. In solid state, there are two enantiomeric molecules that are related by an inversion center as the crystal-building unit (as a translational motif) for the ternary complexes.

Interactions of d(10) metal ions with hippuric acid and cytosine. X-ray structure of the first cadmium (II)-amino acid derivative-nucleobase ternary compound.

The interactions of Zn(II), Cd(II) and Hg(II) with hippuric acid (hipH) were studied and several novel compounds were synthesized and studied by NMR. Some new metal-hippuric-cytosine ternary compounds were formed and the structure of the [Cd(hip)(2)(cyt)(H(2)O)](2) ternary complex resolved. Each cadmium (II) atom has a distorted trigonal bipyramid coordination which is linked to a water molecule, a cytosine via N(3), a carboxylic oxygen atom of a hippurate moiety and two bridging dicoordinated hippurates bound through the carboxylic oxygen atoms. To these five main bonds, two longer ancillary interactions can be observed: the second oxygen of the monocoordinated hippurate group and the carboxylic oxygen of the cytosine ligand. The compound is stabilized by an intramolecular stacking between the benzene and cytosine rings and by the hydrogen bonds between the coordinated water molecules and the ligands. This is, to our knowledge, the first structure of a cadmium-amino acid derivative-natural nucleobase compound described so far.

Uracilato and 5-halouracilato complexes of Cu(II), Zn(II) and Ni(II). X-ray structures of [Cu(uracilato-N(1))(2)(NH(3))(2)].2(H(2)O), [Cu(5-chlorouracilato-N(1))(2)(NH(3))(2)](H(2)O)(2), [Ni(5-chlorouracilato-N(1))(2)(en)(2)].2H(2)O and [Zn(5-chlorouracilato-N(1))(NH(3))(3)].(5-chlorouracilato-N(1)).(H(2)O).

Four new complexes of uracilato and 5-halouracilato with the divalent metal ions Cu(II), Zn(II) and Ni(II) were obtained and structurally characterized. [Cu(uracilato- N(1))(2)(NH(3))(2)].2(H(2)O) (1) and [Cu(5-chlorouracilato-N(1))(2)(NH(3))(2)](H(2)O)(2) (2) complexes present distorted square planar co-ordination geometry around the metal ion. Although an additional axial water molecule is present [Cu(II)-OH(2)=2.89 A (for 1) and 2.52 A (for 2)] in both cases, only in the complex 2 would be considered in the limit of a bond distance. The Zn(II) in [Zn(5-chlorouracilato-N(1))(NH(3))(3)].(5-chlorouracilato-N(1)).(H(2)O) presents a tetrahedral co-ordination with three ammonia molecules and the N(1) of the corresponding uracilato moiety. A non-coordinated uracilato molecule is present as a counterion and a recognition between co-ordinated and free ligands, by means a tandem of H-bonds, should be mentioned. Finally, the complex [Ni(5-chlorouracilato-N(1))(2)(en)(2)] (H(2)O)(2) (where en is ethylenediamine) presents a typical octahedral trans co-ordination with additional hydrogen bonds between 5-chlorouracilato and the NH(2) groups of ethylenediamine units.

Bilateral popliteal artery entrapment syndrome. A case report and clinical considerations.

The authors report their experience in the management of a rare case of bilateral popliteal artery entrapment syndrome occurred to their observation. This unusual pathological entity, arising mostly in young subjects and unilaterally, in this case was bilateral and developed in a 51 years old patient. The clinical case is fully documented by multiple Echo-Doppler and arteriographic examinations which clearly demonstrate the peculiar characteristics of the disease. Finally the patient received a two stage surgical treatment with a good global result. The Authors stress the importance of a correct diagnostic approach to this unusual disease to avoid the possibility to misdiagnose it.

Superior mesenteric and renal artery embolism during PTA and re-stenting of infrarenal abdominal aorta. Report of a case and review of the literature.

The authors report a case of acute superior mesenteric and right renal artery embolism that occurred during an interventional radiological procedure on the abdominal aorta of a young diabetic woman. The onset of a severe abdominal pain during the procedure evoked the clinical suspicion of intestinal ischemia related to the dislodgement of atheroembolic material into the mesenteric artery; the event was correctly diagnosed, but the surgical therapy was delayed by many hours because of the fact that the patient was in a peripheral hospital of the region and had to be transferred to our institution. Fortunately in spite of the considerable delay, the operation was fully successful, probably because of the favourable location of the embolus, which allowed collateral splanchnic circulation to maintain a good metabolic balance.

Left-sided inferior vena cava. Report of a case occasionally encountered while performing an aorto bifemoral bypass and review of the literature.

We present a case of left-sided inferior vena cava unexpectedly observed during an operation of aorto bifemoral bypass in a patient with severe Leriche syndrome and almost complete obstruction of the infrarenal aorta. This very rare congenital malformation (0.2-0.5%) was not recognized by the duplex scanner performed preoperatively, probably because of the low level of suspicion carried on by an experienced operator. AngioCT or angioMR, which would have surely shown us the anomaly, were not done because, in the lack of an aneurysmal disease or other abdominal pathological situations, these investigations were not required before operation. The possible hazards of such an unrecognized malformation are great, mostly in terms of incontrollable intraoperative hemorrhages, but the final outcome of this case was positive.

Left-sided inferior vena cava in patients submitted to aorto iliac surgery. Our experience and review of the literature.

We present two cases of left sided inferior vena cava, one unexpectedly observed during an operation of aorto bifemoral bypass in a patient with severe Leriche syndrome and almost complete obstruction of the infrarenal aorta, the second in a patient with an aneurysm of the abdominal aorta, in whom the anomaly was recognized before the operation. This very rare congenital malformation (0.2-0.5%) was not recognized in the first patient by the duplex scanner performed preoperatively, probably because of the low level of suspicion carried on by an experienced operator. Computer tomography angiography or magnetic resonance angiography, which would have surely shown us the anomaly, were not done in the first patient because, in the lack of an aneurysmal disease or other abdominal pathological situations, these investigations were not required before operation. The possible hazards of such an unrecognized malformation are great, mostly in terms of uncontrollable intraoperative hemorrhages, but the final outcome of this case was positive.

Vagal paraganglioma. Report of a case surgically treated and review of the literature.

The authors report the observation of one case of vagal paraganglioma occurred in a young woman. The tumor manifested itself as a left sub-mandibular tumescence; the very first diagnostic approach was achieved through echography, which showed a mass behind the internal carotid artery and compressing the internal jugular vein. The following examinations, represented by neck CT, NMR, angiography and fine needle aspiration initially directed towards the suspicion of chemodectoma. Only at operation, the anatomical situation of the tumor, which encapsulated the vagus nerve and the subsequent results of the hystological examination revealed the correct diagnosis of vagal paraganglioma.

Aneurysms of the hypogastric artery following surgery of the abdominal aorta. A report of two cases and review of the literature.

Isolated aneurysms of the hypogastric artery are very rare and account for between 0.04% and 0.4% of all intra-abdominal aneurysms. In 85% of cases they are monolateral and are present in association with an aneurysm of the infrarenal abdominal aorta and of the common and external iliac arteries, they make up part of a poly-aneurysmal disease. Unless the patient is an extremely poor condition, surgical treatment is generally indicated for aneurysms greater than 3 cm; close monitoring of those with smaller aneurysms is recommended. Two patients presented with isolated aneurysm of an internal iliac artery which had developed several years after aortoiliac surgery. The one received surgical treatment; the other, who was in poor general conditions and at high risk for surgery, underwent endovascular embolization. Both procedures were successful, with a current follow-up between 15 and 18 months. Endovascular embolization, as performed in the 2(nd) patient, provided an alternative to the surgical procedure. After injection in the aneurysmal sac of the Gianturco spirals, a covered stent was placed in the iliac axis to exclude the inflow of the hypogastric artery. According to our experience of 2 patients, the one treated surgically and the other submitted to a less invasive endovascular procedure, we can state that both methods are practicable. The final choice lies with the vascular surgeon, after weighing the multiple factors that each case involves.

True aneurysm of the inferior thyroid artery. Case report and review of the literature.

In a patient already operated for abdominal aortic aneurysm fifteen months previously, because of the onset of aspecific vertigo, instrumental investigations of the supra aortic vessels showed us the presence of a non palpable mass, in the left side of the neck. Duplex Scanner, angio-CT and Angiography let us suspect the presence of an aneurysm located somewhere in the course of the thyrocervical trunk. At the operation the aneurysm, which we originally suspected to be a false and a possibly iatrogenic one, in the reality was a true aneurysm and was located at the termination of the thyrocervical trunk, just in the first segment of the inferior thyroid artery. The case is reported because of its rarity and the difficulties in the preoperative diagnosis.

Post stenotic aneurysm of the inferior mesenteric artery: case report and discussion.

A case of post stenotic aneurysm of the inferior mesenteric artery (IMA) is reported. This case underlines the importance of a correct diagnosis by instrumental methods such as Duplex Scanner, CT and Arteriography, mainly when the first diagnosis is aneurysm of the abdominal aorta (AAA). Our patient showed, furthermore, the occlusion of the celic axis, of the superior mesenteric artery and of both hypogastric arteries; thus his intestinal vascularization was represented, from the gastric fundus until the rectum, only by the inferior mesenteric artery and its collateral network. The operation consisted in the resection and reimplantation of the vessel on the left side of the terminal abdominal aorta, after removal of the dilated segment. This location was chosen because of the coexistence of a mild dilatation of the aorta of 2.8 cm and considering possible aortic surgery for AAA in the future.