High-Level Expression of Wild-Type p53 in Melanoma Cells is Frequently Associated with Inactivity in p53 Reporter Gene Assays

Background: Inactivation of the p53 pathway that controls cell cycle progression, apoptosis and senescence, has been proposed to occur in virtually all human tumors and p53 is the protein most frequently mutated in human cancer. However, the mutational status of p53 in melanoma is still controversial; to clarify this notion we analysed the largest series of melanoma samples reported to date. Methodology/Principal Findings: Immunohistochemical analysis of more than 180 melanoma specimens demonstrated that high levels of p53 are expressed in the vast majority of cases. Subsequent sequencing of the p53 exons 5–8, however, revealed only in one case the presence of a mutation. Nevertheless, by means of two different p53 reporter constructs we demonstrate transcriptional inactivity of wild type p53 in 6 out of 10 melanoma cell lines; the 4 other p53 wild type melanoma cell lines exhibit p53 reporter gene activity, which can be blocked by shRNA knock down of p53. Conclusions/Significance: In melanomas expressing high levels of wild type p53 this tumor suppressor is frequently inactivated at transcriptional level

Research needs in allergy: an EAACI position paper, in collaboration with EFA

Abstract In less than half a century, allergy, originally perceived as a rare disease, has become a major public health threat, today affecting the lives of more than 60 million people in Europe, and probably close to one billion worldwide, thereby heavily impacting the budgets of public health systems. More disturbingly, its prevalence and impact are on the rise, a development that has been associated with environmental and lifestyle changes accompanying the continuous process of urbanization and globalization. Therefore, there is an urgent need to prioritize and concert research efforts in the field of allergy, in order to achieve sustainable results on prevention, diagnosis and treatment of this most prevalent chronic disease of the 21 st century. The European Academy of Allergy and Clinical Immunology (EAACI) is the leading professional organization in the field of allergy, promoting excellence in clinical care, education, training and basic and translational research, all with the ultimate goal of improving the health of allergic patients. The European Federation of Allergy and Airways Diseases Patients' Associations (EFA) is a non-profit network of allergy, asthma and Chronic Obstructive Pulmonary Disorder (COPD) patients' organizations. In support of their missions, the present EAACI Position Paper, in collaboration with EFA, highlights the most important research needs in the field of allergy to serve as key recommendations for future research funding at the national and European levels. Although allergies may involve almost every organ of the body and an array of diverse external factors act as triggers, there are several common themes that need to be prioritized in research efforts. As in many other chronic diseases, effective prevention, curative treatment and accurate, rapid diagnosis represent major unmet needs. Detailed phenotyping/endotyping stands out as widely required in order to arrange or re-categorize clinical syndromes into more coherent, uniform and treatment-responsive groups. Research efforts to unveil the basic pathophysiologic pathways and mechanisms, thus leading to the comprehension and resolution of the pathophysiologic complexity of allergies will allow for the design of novel patient-oriented diagnostic and treatment protocols. Several allergic diseases require well-controlled epidemiological description and surveillance, using disease registries, pharmacoeconomic evaluation, as well as large biobanks. Additionally, there is a need for extensive studies to bring promising new biotechnological innovations, such as biological agents, vaccines of modified allergen molecules and engineered components for allergy diagnosis, closer to clinical practice. Finally, particular attention should be paid to the difficult-to-manage, precarious and costly severe disease forms and/or exacerbations. Nonetheless, currently arising treatments, mainly in the fields of immunotherapy and biologicals, hold great promise for targeted and causal management of allergic conditions. Active involvement of all stakeholders, including Patient Organizations and policy makers are necessary to achieve the aims emphasized herein

Relationship between low tube voltage (70 kV) and the iodine delivery rate (IDR) in CT angiography: An experimental in-vivo study

Objective Very short acquisition times and the use of low-kV protocols in CTA demand modifications in the contrast media (CM) injection regimen. The aim of this study was to optimize the use of CM delivery parameters in thoraco-abdominal CTA in a porcine model. Materials and methods Six pigs (55–68 kg) were examined with a dynamic CTA protocol (454 mm scan length, 2.5 s temporal resolution, 70 s total acquisition time). Four CM injection protocols were applied in a randomized order. 120 kV CTA protocol: (A) 300 mg iodine/kg bodyweight (bw), IDR = 1.5 g/s (flow = 5 mL/s), injection time (ti) 12 s (60 kg bw). 70 kV CTA protocols: 150 mg iodine/kg bw: (B) IDR = 0.75 g/s (flow = 2.5 mL/s), ti = 12 s (60 kg bw); (C) IDR = 1.5 g/s (flow = 5 mL/s), ti = 12 s (60 kg bw); (D) IDR = 3.0 g/s (flow = 10 mL/s), ti = 3 s (60 kg bw). The complete CM bolus shape was monitored by creating time attenuation curves (TAC) in different vascular territories. Based on the TAC, the time to peak (TTP) and the peak enhancement were determined. The diagnostic window (relative enhancement > 300 HU), was calculated and compared to visual inspection of the corresponding CTA data sets. Results The average relative arterial peak enhancements after baseline correction were 358.6 HU (A), 356.6 HU (B), 464.0 HU (C), and 477.6 HU (D). The TTP decreased with increasing IDR and decreasing ti, protocols A and B did not differ significantly (systemic arteries, p = 0.843; pulmonary arteries, p = 0.183). The delay time for bolus tracking (trigger level 100 HU; target enhancement 300 HU) for single-phase CTA was comparable for protocol A and B (3.9, 4.3 s) and C and D (2.4, 2.0 s). The scan window time frame was comparable for the different protocols by visual inspection of the different CTA data sets and by analyzing the TAC. Conclusions All protocols provided sufficient arterial enhancement. The use of a 70 kV CTA protocol is recommended because of a 50% reduction of total CM volume and a 50% reduced flow rate while maintaining the bolus profile. In contrast to pulmonary arterial enhancement, the systemic arterial enhancement improved only slightly increasing the IDR from 1.5 g/s to 3 g/s because of bolus dispersion of the very short bolus (3s) in the lungs

Relationship between low tube voltage (70 kV) and the iodine delivery rate (IDR) in CT angiography: An experimental <i>in-vivo</i> study - Fig 2

<p><b>Fitted time attenuation curves of the aorta from a representative animal for protocols A, B, C and D.</b> The dotted line at 100 HU represents the bolus tracking threshold level; the dotted line at 300 HU represents the diagnostic target attenuation. The diagnostic window (enhancement > 300 HU) is comparable for all protocols (7–9 s), and the bolus tracking delay time is shorter for protocols C and D than that for protocols A and B.</p

Contrast media protocols.

<p>Contrast media protocols.</p

Effect of the IDR on peak enhancement and time to peak in 70 kV CTA.

<p>Correlation of peak enhancement and time to peak of the pulmonary arteries and aorta (for the different IDR of the CM injection protocols. The diagnostic window (t300 HU) and the bolus tracking delay time are given for the aorta and its main branches.</p

Time-to-peak (s, mean ± standard deviation) for injection protocol A-D.

<p>Time-to-peak (s, mean ± standard deviation) for injection protocol A-D.</p

Peak arterial enhancement (HU, mean ± standard deviation) for injection protocol A-D.

<p>Peak arterial enhancement (HU, mean ± standard deviation) for injection protocol A-D.</p

Relative HU-enhancement within the aorta for the different protocols at the optimal delay time.

<p>Measurements were taken along a centerline of the aorta. The markers indicate the mean, the error-bars the variation between the different animals for each protocol.</p