Plasma total cell-free DNA (cfDNA) is a surrogate biomarker for tumour burden and a prognostic biomarker for survival in metastatic melanoma patients.

INTRODUCTION: Tumour burden is a prognostic biomarker in metastatic melanoma. However, tumour burden is difficult to measure and there are currently no reliable surrogate biomarkers to easily and reliably determine it. The aim of this study was to assess the potential of plasma total cell free DNA as biomarker of tumour burden and prognosis in metastatic melanoma patients. MATERIALS AND METHODS: A prospective biomarker cohort study for total plasma circulating cell-free DNA (cfDNA) concentration was performed in 43 metastatic melanoma patients. For 38 patients, paired blood collections and scan assessments were available before treatment and at first response evaluation. Tumour burden was calculated as the sum of volumes from three-dimensional radiological measurements of all metastatic lesions in individual patients. RESULTS: Baseline cfDNA concentration correlated with pre-treatment tumour burden (ρ = 0.52, P < 0.001). Baseline cfDNA levels correlated significantly with hazard of death and overall survival, and a cut off value of 89 pg/µl identified two distinct prognostic groups (HR = 2.22 for high cfDNA, P = 0.004). Patients with cfDNA ≥89 pg/µl had shorter OS (10.0 versus 22.7 months, P = 0.009; HR = 2.22 for high cfDNA, P = 0.004) and the significance was maintained when compared with lactic dehydrogenase (LDH) in a multivariate analysis. We also found a correlation between the changes of cfDNA and treatment-related changes in tumour burden (ρ = 0.49, P = 0.002). In addition, the ratio between baseline cfDNA and tumour burden was prognostic (HR = 2.7 for cfDNA/tumour volume ≥8 pg/(µl*cm3), P = 0.024). CONCLUSIONS: We have demonstrated that cfDNA is a surrogate marker of tumour burden in metastatic melanoma patients, and that it is prognostic for overall survival.

Molecular mechanisms and modulation of key pathways underlying the synergistic interaction of sorafenib with erlotinib in non-small-cell-lung cancer (NSCLC) cells.

Combination of drugs with different targets is a logical approach to overcome multilevel cross-stimulation among key pathways in NSCLC progression such as EGFR, K-Ras and VEGFR. The sorafenib-erlotinib combination showed clinical activity and acceptable safety. Therefore, we evaluated mechanisms underlying sorafenib-erlotinib interaction in seven NSCLC cell lines selected for their heterogeneous pattern of EGFR and Raf-kinase-inhibitor protein (RKIP) expression, and EGFR/K-Ras mutations. Pharmacologic interaction was studied using MTT/SRB assays and the combination index (CI) method, while effects on EGFR, Erk1/2 and Akt phosphorylation, cell cycle and apoptosis were studied with western-blot, ELISA, and flow cytometry. Intracellular drug concentrations were measured with LC-MS/MS, whereas kinase activity profiles were generated on tyrosine kinase peptide substrate arrays. Synergism was detected in all cell lines, with CIs < 0.6 in K-Ras mutated A549, SW1573 and H460, as well as in H1975 (EGFR-T790M) cells. Sorafenib slowed cell cycle progression and induced apoptosis, which was significantly increased in the combination. Moreover, sorafenib reduced Akt/ERK phosphorylation in erlotinib-resistant cells, associated with significant RKIP up-regulation. No direct drug interaction was detected by LC-MS/MS measurement, while lysates from A549 and H1975 cells exposed to erlotinib+sorafenib showed a significant inhibition in the phosphorylation of 16 overlapping peptides, including sites from RAF, VEGFR2, PDGFR, CDK2 and SRC, suggesting new markers to identify NSCLC patients who are likely to respond to this treatment. In conclusion, several mechanisms, including apoptosis-induction, modulation of expression/phosphorylation of RKIP and crucial kinases contribute to erlotinib-sorafenib synergistic interaction and should be evaluated in future trials for the rational development of this combination in NSCLC.

[Lymphocele after renal transplant: considerations based on a clinical case]. / Il linfocele dopo trapianto renale: considerazioni da un caso clinico.

The Authors study again the problems of lymphocele in kidney transplantation by showing an interesting clinical case: a lymphocele with smallest size, developing an intensively compressive action on iliac vein. In the light of the experience of 354 transplantations, they discuss the etiopathology and clinics thereof. They particularly emphasize the diagnostical role of echography. Whereas the asymptomatic lymphoceles require no treatments for their trend to be reabsorbed, the symptomatic ones need a quick solution of the compressive effect. Then they consider the therapeutical solutions suggested as an alternative to the interperitoneal marsupialization, which remains the most widely used method.

[Significance of mixed lymphocyte culture (MLC) in renal transplants from living donors]. / Significatività del MLC nel trapianto renale da donatore vivente.

Results of MLC were correlated with kidney-graft survival of recipients of living related donor. The 56 patients tested by MLC were divided into two groups according as the stimulation index was more or less than 5. In 20 of these patients transplanted, renal allograft survival correlated better with low stimulation in MLC, suggesting more histocompatibility.

[Relation between cytomegalovirus infection and the so-called intolerance to azathioprine in renal transplantation]. / Rapporto tra infezione da virus citomegalico e la cosiddetta intolleranza all'azatioprina nel trapianto renale.

Renal transplant recipients can develop hepatic function abnormalities or severe leucopenia after transplantation. Generally it is thought to be due to azathioprine intolerance and patients are treated by curtailment of immunosuppressive therapy, being subsequently at risk to lose their allograft because of rejection. Evidence of Cytomegalovirus (CMV) infection is also common after renal transplantation. It is generally thought that the majority of these infections are asymptomatic, but they can be accompanied by leucopenia and/or hepatic function abnormalities. Sixty-nine renal transplant recipients have been studied for at least three months in order to investigate the relationship between CMV and azathioprine intolerance after transplantation. Twenty-five out of 58 patients who underwent seroconversion to CMV (a fourfold or greater rise in titer of CMV antibodies) after transplantation or who had a high CMV titer (greater than or equal to 1 : 16) prior to transplant, developed azathioprine intolerance. None of 11 patients who before renal transplantation had low CMV titers and who did'nt underwent seroconversion did not tolerate azathioprine. Therefore the Authors advance the hypothesis that azathioprine intolerance following renal transplantation can be often due to an asymptomatic and unknown CMV infection.

[Diagnosis and therapy of lymphocele as a complication of renal transplantation]. / Diagnostica e terapie del linfocele quale complicazione del trapianto renale.

Of 165 renal transplanted patients, three (1.8%) developed a pelvic lymphocele. Decreased renal function, leg edema, a lower quadrant abdominal mass and fluid retention represented suspicion as the possibility of lymph collection in the perirenal space. Excretory urography associated with pelvic tomography, Computerized Tomography and B scan ultrasound confirmed diagnosis and were helpful in the post-operative follow-up. Drainage procedure restored normal renal function and morphology. External drainage and marsupialization into the peritoneum have been used successfully.

[Aseptic osseous necrosis after renal transplantation]. / La necrosi ossea asettica dopo trapianto renale.

Of 165 renal Transplantated patients, 12 developed aseptic bone necrosis in the femoral head (6 patients), in the femoral condyle (5 patients), in the astragalus (1 patient). The onset of symptoms was 6 to 23 months after transplantation. 99mTc-O4-MDP bone scintigraphy and radiological examination associated with clinical signs confirmed the diagnosis. Unresolved hyperparathyroidism, phosforus depletion, ponderal increase, total i.v. prednisolone-boluses and trauma represented conditions which might predispose to the development of lesion. 8 patients were managed with conservative treatment. 4 patients required a total of 8 operations: head replacement arthroplasty and articular cartilage reimplant in two patients with disease involving femoral head; articular cartilage reimplant and condyle replacement arthroplasty in two patients with disease involving femoral condyle.

[Internal arteriovenous fistula in the antecubital region]. / La fistola artero-venosa interna in regione antecubitale.

An assessment was made of the anterior cubital region as a vascular approach for primary and alternative dialysis. Reference is made to 36 fistulae in this area within 44 months' survival in the drawing of various conclusions with regard to surgical technique. End anastomosis of the median, cephalic or basilic vein laterally on the brachial or radial artery is recommended as a means of preventing or cutting down the more common complications associated with internal arteriovenous fitulae. The anastomosis should not exceed 6 mm in diameter. Complications led to loss of fistula function. In many cases, however, they did not prevent employment of the region for alternative approaches, such as superficialisation of the basilic vein or prosthesis.

Serum gastrin levels in patients with renal failure on maintenance haemodialysis and after successful kidney transplantation.

Chronic renal failure in man is associated with a hypergastrinaemia which is not corrected by haemodialysis. Serum gastrin concentrations were measured in 66 patients on maintenance haemodialysis and in 27 patients after successful kidney transplantation. The basal gastrin levels distinguished 2 groups of patients on maintenance haemodialysis: a first group with gastrin values less than or equal to 120 pg/ml (mean 58+/-29.88 pg/ml) and a second group with gastrin values greater than 120 pg/ml (mean 295+/-257 pg/ml). The onset of urine output after kidney transplantation was associated with a dramatic fall in gastrin concentration (3 patients studied in the early post-transplant period). 24 patients with stabilized transplant function had serum gastrin levels less than 120 pg/ml (mean 21.33+/- 17.29 pg/ml). The gastrin response to a standard protein meal depended on the basal gastrin concentration and was different in patients with renal failure on maintenance haemodialysis and those with a successful kidney transplantation.