Interruption of p53-MDM2 Interaction by Nutlin-3a in Human Lymphoma Cell Models Initiates a Cell-Dependent Global Effect on Transcriptome and Proteome Level.

In most lymphomas, p53 signaling pathway is inactivated by various mechanisms independent to p53 gene mutations or deletions. In many cases, p53 function is largely regulated by alterations in the protein abundance levels by the action of E3 ubiquitin-protein ligase MDM2, targeting p53 to proteasome-mediated degradation. In the present study, an integrating transcriptomics and proteomics analysis was employed to investigate the effect of p53 activation by a small-molecule MDM2-antagonist, nutlin-3a, on three lymphoma cell models following p53 activation. Our analysis revealed a system-wide nutlin-3a-associated effect in all examined lymphoma types, identifying in total of 4037 differentially affected proteins involved in a plethora of pathways, with significant heterogeneity among lymphomas. Our findings include known p53-targets and novel p53 activation effects, involving transcription, translation, or degradation of protein components of pathways, such as a decrease in key members of PI3K/mTOR pathway, heat-shock response, and glycolysis, and an increase in key members of oxidative phoshosphorylation, autophagy and mitochondrial translation. Combined inhibition of HSP90 or PI3K/mTOR pathway with nutlin-3a-mediated p53-activation enhanced the apoptotic effects suggesting a promising strategy against human lymphomas. Integrated omic profiling after p53 activation offered novel insights on the regulatory role specific proteins and pathways may have in lymphomagenesis.

Incidence, time trends and survival patterns of childhood pilocytic astrocytomas in Southern-Eastern Europe and SEER, US.

Pilocytic astrocytomas (PA) comprise the most common childhood central nervous system (CNS) tumor. Exploiting registry-based data from Southern and Eastern Europe (SEE) and SEER, US, we opted to examine incidence, time trends, survival and tentative outcome disparities of childhood PA by sociodemographic and clinical features. Childhood PA were retrieved from 12 SEE registries (N = 552; 1983-2014) and SEER (N = 2723; 1973-2012). Age-standardized incidence rates (ASR) were estimated and survival was examined via Kaplan-Meier and Cox regression analysis. ASR of childhood PA during 1990-2012 in SEE was 4.2/106, doubling in the USA (8.2/106). Increasing trends, more prominent during earlier registration years, were recorded in both areas (SEE: +4.1 %, USA: +4.6 %, annually). Cerebellum comprised the most common location, apart from infants in whom supratentorial locations prevailed. Age at diagnosis was 1 year earlier in SEE, whereas 10-year survival was 87 % in SEE and 96 % in SEER, improving over time. Significant outcome predictors were age <1 year at diagnosis diagnosis (hazard ratio, HR [95% confidence intervals]: 3.96, [2.28-6.90]), female gender (HR: 1.38, [1.01-1.88]), residence in SEE (HR: 4.07, [2.95-5.61]) and rural areas (HR: 2.23, [1.53-3.27]), whereas non-cerebellar locations were associated with a 9- to 12-fold increase in risk of death. The first comprehensive overview of childhood PA epidemiology showed survival gains but also outcome discrepancies by geographical region and urbanization pointing to healthcare inequalities. The worse prognosis of infants and, possibly, females merits further consideration, as it might point to treatment adjustment needs, whereas expansion of systematic registration will allow interpretation of incidence variations.

Childhood central nervous system tumour mortality and survival in Southern and Eastern Europe (1983-2014): Gaps persist across 14 cancer registries.

AIM: Childhood central nervous system (CNS) tumour registration and control programs in Southern and Eastern Europe remain thin, despite the lethal nature of the disease. Mortality/survival data were assembled to estimate the burden of malignant CNS tumours, as well as the potential role of sociodemographic survival determinants across 14 cancer registries of this region. METHODS: Average age-adjusted mortality rates were calculated, whereas time trends were quantified through Poisson and Joinpoint regressions. Kaplan-Meier curves were derived for the maximum and the more recent (10 and 5 year) registration periods. Multivariate Cox regression models were used to assess demographic and disease-related determinants. RESULTS: Variations in mortality (8-16 per million) and survival (5-year: 35-69%) were substantial among the participating registries; in most registries mortality trend was stable, whereas Bulgaria, having the highest starting rate, experienced decreasing annual mortality (-2.4%, p=0.001). A steep decrease in survival rates was evident before the second year of follow-up. After controlling for diagnostic subgroup, age, gender and diagnostic year, Greece seemed to present higher survival compared with the other contributing registries, although the follow-up period was short. Irrespective of country, however, rural residence was found to impose substantial adverse repercussions on survival (hazard ratio (HR): 1.2, 95% confidence interval (CI): 1.1-1.4). CONCLUSION: Cross-country mortality and survival variations possibly reflect suboptimal levels of health care delivery and cancer control in some regions of Southern and Eastern Europe, notwithstanding questionable death certification patterns or follow-up procedures. Continuous childhood cancer registration and linkage with clinical data are prerequisite for the reduction of survival inequalities across Europe.

The Digital Microscopy in Organ Transplantation: Ergonomics of the Tele-Pathological Evaluation of Renal and Liver Grafts.

AIM: Ergonomics of Digital Microscopy (DM) based on virtual slides, on Telemedicine Systems (TS) for Tele-Pathological (TPE) evaluation of the grafts (G) in organ transplantation (OT). MATERIAL AND METHODS: Simulation of DM on TS based TPE by 2 specialists on a total of 238 human Renal Graft (RG) and 172 Liver Graft (LG) tissues digital microscopic images diagnosing inflammatory and neoplastic lesions on four different electronic spaces (ES). RESULTS: DM on TS for TPE in OT is elaborated perfectly on the ES of a Desktop, followed by the ES of the applied Exp.-TS. Tablet and Mobile-Phone ES seem significantly risky for the application of DM in OT (p<.001). CONCLUSION: Integration of DM on TS for TPE is feasible, while ergonomics of post-grafting and pre-transplant decision support and planning depend on the size of the working ES.

FAK/Src family of kinases: protective or aggravating factor for ischemia reperfusion injury in nervous system?

INTRODUCTION: The focal adhesion kinase (FAK) and the Src families of kinases are subfamilies of the non-receptor protein tyrosine kinases. FAK activity is regulated by gene amplification, alternative splicing and phosporylation/dephosphorylation. FAK/Src complex has been found to participate through various pathways in neuronal models of ischemia-reperfusion injury (IRI) with conflicting results. The aim of the present review is to summarize the currently available data on this subject. AREAS COVERED: The MEDLINE/PubMed database was searched for publications with the medical subject heading IRI and FAK and/or Src, nervous system. We restricted our search till 2014. We identified 93 articles that were available in English as abstracts or/and full-text articles that were deemed appropriate for our review. EXPERT OPINION: FAK has been found to have a beneficial preconditioning effect on IRI through activation via the protein kinase C (PKC) pathway by anesthetic agents. Of great importance are the interactions between FAK/Src and VEGF that has been already detected as a protective mean for IRI. The effect of VEGF administration might depend on dose as well as on time of administration. A Ca(2+)/calmodulin-dependent protein kinase II or PKC inhibitors seem to have protective effects on IRI by inhibiting ion channels activation.

Catheter based inhibition of arterial calcification by bisphosphonates in an experimental atherosclerotic rabbit animal model.

BACKGROUND: Vascular calcification is an active process, sharing common molecular mechanisms with bone formation. Bisphosphonates are components, which inhibit calcification. The aim of the present study was to evaluate the safety and effectiveness of local delivery of the bisphosphonate zoledronate on inhibition of calcium formation in the arterial wall in an experimental animal model. METHODS: Sixteen New Zealand rabbits were placed on vitamin D enriched atherogenic diet for 3 weeks. Subsequently, all animals underwent angiography of abdominal aorta and common iliac arteries. A mixture containing 500 µg/l zoledronate was delivered on the vascular wall of the target iliac artery, using a dedicated balloon catheter. A placebo mixture was administered on the contralateral iliac artery of each animal, which was used as control. At 28 days all animals were sacrificed. Histologic sections of each common iliac artery were stained with hematoxylin-eosin and von Kossa. Computer-assisted histomorphometry was performed for the calcium content quantification of each section from the target and the control iliac artery. RESULTS: In all animals the local delivery of zoledronate and placebo mixtures was successful and uncomplicated. The mean percentage of the calcium content of the media was higher in the control artery segments compared to the target (2.66 ± 0.73 versus 1.08 ± 0.62 % of the area of the media, p<0.01). CONCLUSIONS: Inhibition of vascular calcification by local catheter-based delivery of bisphosphonate zolendronic acid is effective without evident short-term complications. These finding and its potential clinical implication remain to be confirmed in human studies.

Integrated Tele-radiological and Tele-pathological Evaluation of Pulmonary Grafts for Reducing the Rate of Injured and Diseased Grafts, in Lung Transplant.

AIM: Feasibility and reliability assessment of the integrated Tele-Radiological (TRE) and Tele-Pathological (TPE) evaluation of the Pulmonary Graft (PG) on Telemedicine Systems, in Lung Transplant (LT). MATERIAL AND METHODS: Sensitivity-specificity by simulating TRE of PG on 15 MR thoracic images by two radiologists and TPE by 20 specialists based on a total of 100 human PG digital images. RESULTS: Integration of TRE and TPE of PG showed an accuracy of 94.1% for hematomas and 97.1% for neoplastic identification. CONCLUSION: Integrated TRE and TPE of PG is feasible and reliable for reducing the rate of injured and diseased PG in LT.

Telemedicine systems in organ transplantation: a feasibility and reliability study of the integrated teleradiological and tele-pathological evaluation of the cardiac graft.

UNLABELLED: Aim. Feasibility and reliability of the integrated Tele-Radiological (TRE) and Tele-Pathological (TPE) evaluation of the Cardiac Graft (CG) on Telemedicine Systems. MATERIAL AND METHODS: Sensitivity-specificity analysis by a simulation of TRE of CG on 15 MR thoracic images by two radiologists and of TPE by 20 specialists based on total 100 human CG digital images, assessing injuries/trauma and lesions. RESULTS: Integration of TRE and TPE of CG for remote diagnosis showed an accuracy for: injuries/trauma/hematoma=90.9-100%, and non-neoplastic lesions=94.3%. CONCLUSION: Integration of TRE and TPE of CG is feasible and reliable for Heart Pre-Grafting and Pre-Transplant decision support and planning.

Increased expression of phosphorylated NBS1, a key molecule of the DNA damage response machinery, is an adverse prognostic factor in patients with de novo myelodysplastic syndromes.

The expression of activated forms of key proteins of the DNA damage response machinery (pNBS1, pATM and γH2AX) was assessed by means of immunohistochemistry in bone marrow biopsies of 74 patients with de novo myelodysplastic syndromes (MDS) and compared with 15 cases of de novo acute myeloid leukemia (AML) and 20 with reactive bone marrow histology. Expression levels were significantly increased in both MDS and AML, compared to controls, being higher in high-risk than in low-risk MDS. Increased pNBS1 and γH2AX expression possessed a significant negative prognostic impact for overall survival in MDS patients, whereas pNBS1 was an independent marker of poor prognosis.

Telepathology as a method to optimize quality in organ transplantation: a feasibility and reliability study of the virtual benching of renal graft.

AIM: To search feasibility and reliability of Telemedicine Systems (TS) in the procurement of Renal Transplantation. MATERIAL AND METHODS: By an experimental TS, twenty six (n=26) specialists based on a hundred and thirty (N=130) simulated remote image examinations, assessed sensitivity-specificity of the remote examination of Renal Graft (RG) about diseases and damages (Virtual Benching=VB). RESULTS: Analysis showed: injuries (sensitivity=100%), injuries of the capsula (sensitivity=100%), hematomas (sensitivity=96.2%), non neoplasmatic diseases (specificity=100%), tumors (sensitivity=100%). CONCLUSION: The VB of RG after retrieval is feasible and reliable for prevention of damaged renal RG from being sent to the recipient hospital and for transplant planning.

Telepathology as a method to optimize quality in organ transplantation: a feasibility and reliability study of the virtual benching of pancreas graft.

AIM: To search feasibility and reliability of Telemedicine Systems (TS) in the procurement of Pancreas Transplantation. MATERIAL AND METHODS: By an experimental TS, twenty six (n=26) specialists based on a hundred and thirty (N=130) simulated remote image examinations, assessed sensitivity-specificity of the remote examination of Pancreas Graft (PG) about diseases and damages (Virtual Benching=VB). RESULTS: Analysis showed: Injuries (sensitivity=100%), injuries of the capsula (sensitivity=84.6%), hematomas (sensitivity=92.3%), non neoplasmatic diseases (specificity=92.3%), tumors (sensitivity=72%). CONCLUSION: The VB of PG after retrieval is feasible and reliable for prevention of damaged PG from being sent to the recipient hospital and for transplant planning.

Telepathology as a method to optimize quality in organ transplantation: a feasibility and reliability study of the virtual benching of liver graft.

AIM: To search feasibility and reliability of Telemedicine Systems (TS) in the procurement of Liver Transplantation. MATERIAL AND METHODS: By an experimental TS, twenty six (n=26) specialists based on a hundred and thirty (N=130) simulated remote image examinations, assessed sensitivity-specificity of the remote examination of Liver Graft (LG) about diseases and damages (Virtual Benching=VB). RESULTS: Analysis showed: injuries (sensitivity=96%), injuries of the capsula (sensitivity=91.7%), hematomas (sensitivity=91,7%), non neplasmatic diseases (specificity=100%), tumors (sensitivity=100%). CONCLUSION: The VB of LG after retrieval is feasible and reliable for prevention of damaged liver grafts from being sent to the recipient hospital and for transplant planning.

Telemedicine systems in organ transplantation: a feasibility and reliability study of the integrated teleradiological and tele-pathological evaluation of renal graft.

AIM: To determine feasibility and reliability of the integrated Tele-Radiological (TRE) and Tele-Pathological (TPE) evaluation of the Renal Graft (RG) on Telemedicine Systems (TS). MATERIAL AND METHODS: A sensitivity-specificity analysis by a simulation of the TRE of RG on 15 MR abdominal images by a radiologist and of the TPE of RG by 26 specialists based on 130 human RG images assessing damages and lesions. RESULTS: The integrated analysis of TRE and TPE of RG showed: Sensitivity=96.7%, Specificity=100% and Accuracy=97.6%. CONCLUSION: The TRE integrated with the TPE of RG is feasible and reliable for RG transplant decision support and planning.

Telemedicine systems in organ transplantation: a feasibility and reliability study of the integrated teleradiological and tele-pathological evaluation of pancreas graft.

AIM: To determine feasibility and reliability of the integrated Tele-Radiological (TRE) and Tele-Pathological (TPE) evaluation of the Pancreas Graft (PG) on Telemedicine Systems (TS). MATERIAL AND METHODS: A sensitivity-specificity analysis by a simulation of the TRE of PG on 15 MR abdominal images by a radiologist and of the TPE of PG by 26 specialists based on 130 human PG images assessing damages and lesions. RESULTS: The integrated analysis of TRE and TPE of PG showed: Sensitivity=93.3%, Specificity=100% and Accuracy=95.1%. CONCLUSION: The TRE integrated with the TPE of PG is feasible and reliable for PG transplant decision support and planning.

Telemedicine systems in organ transplantation: a feasibility and reliability study of the integrated teleradiological and tele-pathological evaluation of liver graft.

AIM: To determine feasibility and reliability of the integrated Tele-Radiological (TRE) and Tele-Pathological (TPE) evaluation of the Liver Graft (LG) on Telemedicine Systems (TS). MATERIAL AND METHODS: A sensitivity-specificity analysis by a simulation of the TRE of LG on 15 MR abdominal images by a radiologist and of the TPE of LG by 26 specialists based on 130 human LG images assessing damages and lesions. RESULTS: The integrated analysis of TRE and TPE of LG showed: Sensitivity=96.6%, Specificity=100% and Accuracy=97.5%. CONCLUSION: The TRE integrated with the TPE of LG is feasible and reliable for LG transplant decision support and planning.

Children exposure to trace levels of heavy metals at the north zone of Kifissos River.

This study evaluates the exposure level of primary school children at three different towns in northern Attica, near the banks of the Kifissos River, defining referential values of toxic heavy metals. Concentrations of five toxic metals (arsenic, cadmium, mercury, lead and nickel) were analyzed in water samples from the Kifissos River as well as in the scalp hair of children aged 11 to 12 years old living in the study area. Chronic low-level toxicity of lead and cadmium came into view in recent years as a problem of our civilization. Exposure to concentrations of arsenic (As), cadmium (Cd), mercury (Hg) and lead (Pb) poses a potential threat to humans and can cause effects on children's renal and dopaminergic systems without clear evidence of a threshold, a fact that reinforces the need to control all the potential heavy metal emissions into the environment in order to protect children's health. The results were analyzed with the IBM SPSS Statistics 20 for Windows. The possible influence of sex and area was examined. Statistical differences were observed by t-test between the log-transformed hair concentrations of lead (p=0.021), arsenic (p=0.016) and nickel (p=0.038) in children's scalp hair from the municipalities of Kifisia and Kryoneri. ANOVA one-way test confirmed the difference of Pb concentration in hair between girls and boys from the municipality of Kifisia (p=0.038). The t-test confirms the difference of heavy metal concentrations in river samples between the municipalities Kifisia and Philadelphia in comparison with the samples from Kryoneri. The observations suggest that children living at the municipality of Kifisia are exposed to higher concentrations of heavy metals than the others. Despite all the confounding factors, hair can be used as a biomarker in order to determine the exposure to heavy metals, according to standardized protocols.

Early-stage gastric MALT lymphoma: is it a truly localized disease?

Early-stage gastric mucosa-associated lymphoid tissue lymphoma (GML) is considered a localized disease with an indolent course. Circulating malignant cells have been detected in other early-stage indolent lymphomas by molecular methods. We investigated the incidence of occult blood disease in early-stage GML patients, its impact on clinical outcome, and the similarity between blood and gastric lymphocytic clones. Sixty-two patients with localized GML were included in the study; 51 of them had Helicobacter pylori infection. Monoclonality was investigated by leader polymerase chain reaction. Sequencing was performed for the immunoglobulin variable gene (VH) analysis. Blood involvement was absent in all patients by conventional staging methods. In the whole group of 62 patients, the incidence of blood IgH rearrangement was 45%, and this did not correlate with baseline patient characteristics. The monoclonal blood and gastric products of five patients were sequenced and compared with each other. Clonal identity was evident in four of five patients. The VH3 gene was the most frequently used, both in the blood and in the stomach. Early-stage GML is not a truly localized disease because half the patients had a circulating clone, probably identical to the gastric one. The clinical significance of occult blood disease and the potential appropriate intervention need to be further investigated.

In vivo aortic valve thermal heterogeneity in patients with nonrheumatic aortic valve stenosis the: first in vivo experience in humans.

OBJECTIVES: We investigated in vivo in aortic valve stenosis (AVS) whether there is: 1) thermal heterogeneity within the valve leaflets; 2) temperature difference between the leaflets and the ascending aortic wall; and 3) a possible correlation between heat production, inflammation, and neoangiogenesis. BACKGROUND: Histological studies have demonstrated a potential role of inflammation and neoangiogenesis in AVS. METHODS: We examined 96 leaflets scheduled for aortic valve replacement. Twenty-five patients had AVS, and 7 had aortic valve insufficiency (AVI). Temperature measurements were performed right before hypothermic cardioplegia. Temperature difference (DeltaT) was assigned as the mean temperature of each leaflet minus the temperature of the aortic wall. Histological, immunohistological analysis, and vascular endothelial growth factor (VEGF) immunoreactivity was performed. RESULTS: Significant thermal heterogeneity was recorded within the leaflets of AVS, compared with AVI (1.52 +/- 1.35 degrees C vs. 0.13 +/- 0.11 degrees C, p < 0.01). In AVS DeltaT was greater in all leaflets compared with the AVI group (p < 0.01). Leaflets of AVS had increased inflammatory cell infiltration, calcium deposit, and anti-VEGF expression compared with AVI (p < 0.01). CONCLUSIONS: Thermal heterogeneity is increased in AVS and correlates with inflammatory mononuclear cell infiltration, expression of pro-inflammatory cytokines and neoangiogenic factors.