Integrative molecular profiling of routine clinical prostate cancer specimens.

BACKGROUND: Comprehensive molecular profiling led to the recognition of multiple prostate cancer (PCa) molecular subtypes and driving alterations, but translating these findings to clinical practice is challenging. PATIENTS AND METHODS: We developed a formalin-fixed paraffin-embedded (FFPE) tissue compatible integrative assay for PCa molecular subtyping and interrogation of relevant genetic/transcriptomic alterations (MiPC). We applied MiPC, which combines capture-based next generation sequencing and quantitative reverse transcription PCR (qRT-PCR), to 53 FFPE PCa specimens representing cases not well represented in frozen tissue cohorts, including 8 paired primary tumor and lymph node metastases. Results were validated using multiplexed PCR based NGS and Sanger sequencing. RESULTS: We identified known and novel potential driving, somatic mutations and copy number alterations, including a novel BRAF T599_V600insHT mutation and CYP11B2 amplification in a patient treated with ketoconazole (a potent CYP11B2 inhibitor). qRT-PCR integration enabled comprehensive molecular subtyping and provided complementary information, such as androgen receptor (AR) target gene module assessment in advanced cases and SPINK1 over-expression. MiPC identified highly concordant profiles for all 8 tumor/lymph node metastasis pairs, consistent with limited heterogeneity amongst driving events. MiPC and exome sequencing were performed on separately isolated conventional acinar PCa and prostatic small cell carcinoma (SCC) components from the same FFPE resection specimen to enable direct comparison of histologically distinct components. While both components showed TMPRSS2:ERG fusions, the SCC component exclusively harbored complete TP53 inactivation (frameshift variant and copy loss) and two CREBBP mutations. CONCLUSIONS: Our results demonstrate the feasibility of integrative profiling of routine PCa specimens, which may have utility for understanding disease biology and enabling personalized medicine applications.

Targeting the PI3K/Akt/mTOR signaling pathway in B-precursor acute lymphoblastic leukemia and its therapeutic potential.

B-precursor acute lymphoblastic leukemia (B-pre ALL) is a malignant disorder characterized by the abnormal proliferation of B-cell progenitors. The prognosis of B-pre ALL has improved in pediatric patients, but the outcome is much less successful in adults. Constitutive activation of the phosphatidylinositol 3-kinase (PI3K), Akt and the mammalian target of rapamycin (mTOR) (PI3K/Akt/mTOR) network is a feature of B-pre ALL, where it strongly influences cell growth and survival. RAD001, a selective mTORC1 inhibitor, has been shown to be cytotoxic against many types of cancer including hematological malignancies. To investigate whether mTORC1 could represent a target in the therapy of B-pre ALL, we treated cell lines and adult patient primary cells with RAD001. We documented that RAD001 decreased cell viability, induced cell cycle arrest in G0/G1 phase and caused apoptosis in B-pre ALL cell lines. Autophagy was also induced, which was important for the RAD001 cytotoxic effect, as downregulation of Beclin-1 reduced drug cytotoxicity. RAD001 strongly synergized with the novel allosteric Akt inhibitor MK-2206 in both cell lines and patient samples. Similar results were obtained with the combination CCI-779 plus GSK 690693. These findings point out that mTORC1 inhibitors, either as a single agent or in combination with Akt inhibitors, could represent a potential therapeutic innovative strategy in B-pre ALL.

Non-enhanced MR angiography of renal arteries: comparison with contrast-enhanced MR angiography.

BACKGROUND: The main causes of renal artery stenosis (RAS) are atherosclerosis and fibromuscular dysplasia. Despite contrast-enhanced magnetic resonance angiography (CE-MRA) being a safe and reliable method for diagnosis of RAS especially in young individuals, recently it has been possible to adopt innovative technologies that do not require paramagnetic contrast agents. PURPOSE: To assess the accuracy of steady-state free-precession (SSFP) non-contrast-enhanced magnetic resonance angiography (NC-MRA) by using a 1.5 T MR scanner for the detection of renal artery stenosis, in comparison with breath-hold CE-MRA as the reference standard. MATERIAL AND METHODS: Sixty-three patients (33 men, 30 women) with suspected renovascular hypertension (RVHT) were examined by a 1.5T MR scanner; NC-MRA with an electrocardiography (ECG)-gated SSFP sequence was performed in 58.7% (37/63) of patients; in 41.3% (26/63) of patients a respiratory trigger was used in addition to cardiac gating. CE-MRA, with a three-dimensional gradient echo (3D-GRE) T1-weighted sequence, was performed in all patients within the same session. Maximum intensity projection (MIP) image quality, number of renal arteries, and the presence of stenosis were assessed by two observers (independently for NC-MRA and together for CE-MRA). The agreement between NC-MRA and CE-MRA as well as the inter-observer reproducibility were calculated with Bland-Altman plots. RESULTS: MIP image quality was considered better for NC-MRA. NC-MRA identified 143 of 144 (99.3%) arteries detected by CE-MRA (an accessory artery was not identified). Fourteen stenoses were detected by CE-MRA (11 atherosclerotic, 3 dysplastic) with four of 14 (28.5%) significant stenosis. Bland-Altman plot demonstrated an excellent concordance between NC-MRA and CE-MRA; particularly, the reader A evaluated correctly all investigated arteries, while over-estimation of two stenoses occurred for reader B. Regarding NC-MRA, inter-observer agreement was excellent. CONCLUSION: NC-MRA is a valid alternative to CE-MRA for the assessment of renal arteries.

Cytotoxic activity of the novel Akt inhibitor, MK-2206, in T-cell acute lymphoblastic leukemia.

T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive neoplastic disorder arising from T-cell progenitors. T-ALL accounts for 15% of newly diagnosed ALL cases in children and 25% in adults. Although the prognosis of T-ALL has improved, due to the use of polychemotherapy schemes, the outcome of relapsed/chemoresistant T-ALL cases is still poor. A signaling pathway that is frequently upregulated in T-ALL, is the phosphatidylinositol 3-kinase/Akt/mTOR network. To explore whether Akt could represent a target for therapeutic intervention in T-ALL, we evaluated the effects of the novel allosteric Akt inhibitor, MK-2206, on a panel of human T-ALL cell lines and primary cells from T-ALL patients. MK-2206 decreased T-ALL cell line viability by blocking leukemic cells in the G(0)/G(1) phase of the cell cycle and inducing apoptosis. MK-2206 also induced autophagy, as demonstrated by an increase in the 14-kDa form of LC3A/B. Western blotting analysis documented a concentration-dependent dephosphorylation of Akt and its downstream targets, GSK-3α/ß and FOXO3A, in response to MK-2206. MK-2206 was cytotoxic to primary T-ALL cells and induced apoptosis in a T-ALL patient cell subset (CD34(+)/CD4(-)/CD7(-)), which is enriched in leukemia-initiating cells. Taken together, our findings indicate that Akt inhibition may represent a potential therapeutic strategy in T-ALL.

Volumetric analysis of the aneurysmal sac with computed tomography in the follow-up of abdominal aortic aneurysms after endovascular treatment.

PURPOSE: Our aim was to assess the usefulness of volumetric analysis for the follow-up of abdominal aortic aneurysms after endovascular repair (EVAR) and operator independence of the method. MATERIALS AND METHODS: We retrospectively evaluated 99 computed tomography (CT) exams of 33 patients. Two blinded operators assessed the volume before treatment and after EVAR at 1-3 and 12-24 months. Friedman's statistical test was used to assess the reproducibility of the method. The time required for postprocessing by the two operators was compared. RESULTS: One patient was excluded. Twenty-one patients showed no endoleak: 12/21 showed a volume reduction at both follow-up scans (9.7% and 19.5%, respectively); 8/21 showed an early volume increase (9.8%) with a late reduction (10.5%); 1/21 patient showed a volume increase at both follow-up scans (endotension). Eleven patients had an endoleak (one type I, nine type II and one type III); 4/9 type II endoleaks showed a volume reduction at both post-EVAR scans (8.5% and 19.5%). All other cases showed a volume increase after EVAR (type II 15.4%/16.8%, type I 24.1%/9.1%, type III 8%/10.7%). The Friedman statistical test assessed operator independence with p < 0.001. Mean difference between the two operators was 0.9% (0-4.3%). CONCLUSIONS: CT volume analysis is an accurate and reproducible modality for the follow-up of abdominal aortic aneurysms after EVAR. At early follow-up, contrast-enhanced CT remains mandatory to identify small endoleaks. For later follow-up, volumetric analysis would eliminate the need for contrast material in asymptomatic patients with stable or decreasing aneurysm volume.

Endovascular treatment of ruptured abdominal aortic aneurysms: aorto-uni-iliac or bifurcated endograft?

PURPOSE: This study evaluated the safety and technical and clinical success rates of positioning endovascular endografts (EG) in ruptured abdominal aneurysms. MATERIALS AND METHODS: Patients with a ruptured abdominal aortic aneurysm confirmed by contrast-enhanced computed tomography angiography (CTA) were eligible for the analysis. Of 67 patients, 42 (62.7%) were treated with EG. Thirteen patients (30.9%) received an aorto-uni-iliac EG (group A) and 29 a bifurcated EG (group B). Patients were divided for comparative analysis according to the configuration of the EG implanted. RESULTS: The primary technical success rate was 100%; the primary clinical success rate was 95% (40/42). There were two intraoperative deaths (4.7%) related to intractable shock. No patient required conversion to open repair. Overall, 12 patients (28.5%) died within 30 days. The in-hospital death rate was 30.9% (13/42). Hospital mortality rate was statistically higher in group A; the type of EG and intensive care unit admission were the only independent predictors of hospital mortality. CONCLUSIONS: In our experience, a higher mortality rate was observed for the aorto-uni-iliac configuration; shock at admission was confirmed as the most important factor for postoperative survival.

Comparison between MRI with spin-echo echo-planar diffusion-weighted sequence (DWI) and histology in the diagnosis of soft-tissue tumours.

PURPOSE: Our aim was to assess the usefulness of magnetic resonance imaging (MRI) with spin-echo echo-planar diffusion-weighted sequences (SE-EPI-DWI) in the study of primary and secondary soft-tissue tumours by correlating the results of imaging and histology. MATERIAL AND METHODS: We retrospectively studied 23 patients (14 men, 9 women; age range 25-87 years) affected by soft-tissue lesions. The MRI study was performed with baseline and contrast-enhanced SE-T1, proton density/T2-weighted (PD/T2), fat-saturated (FATSAT) DP/T2 and single-shot SE-EPI-DWI (b value 50-400- 800s/mm2) sequences. RESULTS: We identified 7/23 benign lesions (three myxoid, four nonmyxoid) and 16/23 malignant tumours (four myxoid, 12 nonmyxoid) with a mean diameter between 21 mm and 20 cm. Qualitative analysis of DWI showed persistence of high signal intensity for increasing b-values in all malignant tumours. Quantitative DWI analysis of the apparent diffusion coefficient (ADC) maps showed a statistical difference between benign and malignant lesions. CONCLUSIONS: In our experience, DWI with qualitative and quantitative analysis correlated well with histology.

Recurrence of intracardiac large B-cell non-Hodgkin's lymphoma (NHL) encasing and nearly obliterating the right coronary artery: a case report.

Among malignant tumors of the heart, primary malignant lymphomas are unusual and they are typically non-Hodgkin's B-cell cancers. A 73-year-old man affected by non-Hodgkin lymphoma (NHL) treated with four cycle of chemotherapy and radiotherapy was admitted to the Emergency Department for chest pain. Echocardiography showed a mass inside the right ventricle obstructing blood outflow in the pulmonary artery. The ECG-gated angio-multidetector computed tomography (MDCT) examination confirmed a solid mass in the right ventricle encasing the proximal-middle tract of the right coronary artery (RCA); RCA stenosis was confirmed by coronary angiography. After trans-thoracic CT-guided biopsy the mass was characterized as a recurrence of NHL and the patient started a new cycle of chemotherapy. After 15 days a MDCT exam showed both mass reduction and absence of RCA significant stenosis. MDCT imaging allows an accurate assessment of tumour extension and it represents an useful guide during biopsy procedures, necessary for a precise histological characterization of neoplasms.

MDCT evaluation of the cardiac venous system.

PURPOSE: This study was undertaken to evaluate the usefulness of electrocardiographically (ECG)-gated multidetector-row computed tomography (MDCT) for the assessment of the coronary venous system and detection of its anatomical variants, in order to identify those suitable for lead placement in cardiac resynchronisation therapy (CRT). MATERIALS AND METHODS: We retrospectively examined the coronary MDCT studies of 89 patients (73 males, 16 females, average age 62.5 years, range 31-79) referred for suspected coronary artery disease. The cardiac venous system was assessed in all patients using three-dimensional (3D) postprocessing on a dedicated Vitrea workstation (five patients were excluded from the analysis). RESULTS: The coronary sinus, the great cardiac vein, the anterior interventricular vein and the middle cardiac vein were visualised in all cases. The lateral cardiac vein was visualised in 56/84 patients (67%) and the posterior cardiac vein in 63/84 patients (75%), never both missing. Along the postero-lateral wall of the left ventricle, only one branch was present in 44 cases, two branches in 21 cases and three or more branches in 19/84 cases (22%). Evaluation of the maximum diameter revealed that the lateral vein was dominant over the posterior vein in 20/40 cases. The small cardiac vein was visualised in 11/84 cases. CONCLUSIONS: MDCT provides good depiction of the cardiac venous system, enabling the study of the vessel course and the identification of anatomical variants. Hence, this imaging technique could be proposed for the preoperative planning of CRT in selected patients.

[The management of head trauma in a general surgery department. a contribution to an epidemiological analysis]. / La gestione del trauma cranico in un reparto di chirurgia generale. Contributo per un'analisi epidemiologica.

Personal experience in the management of head injury in a General Surgery Department is reported and a series of 352 patients analysed with a view to identifying variations in the epidemiological profile of this pathology over the last decade and in order to establish indications for admission and the transfer of the patient to specialist departments.