Preliminary Evidence on the Diagnostic and Molecular Role of Circulating Soluble EGFR in Non-Small Cell Lung Cancer

Assessment of biological diagnostic factors providing clinically-relevant information to guide physician decision-making are still needed for diseases with poor outcomes, such as non-small cell lung cancer (NSCLC). Epidermal growth factor receptor (EGFR) is a promising molecule in the clinical management of NSCLC. While the EGFR transmembrane form has been extensively investigated in large clinical trials, the soluble, circulating EGFR isoform (sEGFR), which may have a potential clinical use, has rarely been considered. This study investigates the use of sEGFR as a potential diagnostic biomarker for NSCLC and also characterizes the biological function of sEGFR to clarify the molecular mechanisms involved in the course of action of this protein. Plasma sEGFR levels from a heterogeneous cohort of 37 non-advanced NSCLC patients and 54 healthy subjects were analyzed by using an enzyme-linked immunosorbent assay. The biological function of sEGFR was analyzed in vitro using NSCLC cell lines, investigating effects on cell proliferation and migration. We found that plasma sEGFR was significantly decreased in the NSCLC patient group as compared to the control group (median value: 48.6 vs. 55.6 ng/mL respectively; p = 0.0002). Moreover, we demonstrated that sEGFR inhibits growth and migration of NSCLC cells in vitro through molecular mechanisms that included perturbation of EGF/EGFR cell signaling and holoreceptor internalization. These data show that sEGFR is a potential circulating biomarker with a physiological protective role, providing a first approach to the functional role of the soluble isoform of EGFR. However, the impact of these data on daily clinical practice needs to be further investigated in larger prospective studies.     

Solution conformation of the Pseudomonas syringae MSU 16H phytotoxic lipodepsipeptide Pseudomycin A determined by computer simulations using distance geometry and molecular dynamics from NMR data

Pseudomycin A is a cyclic lipodepsinonapeptide phytotoxin produced by a strain of the plant pathogenic bacterium Pseudomonas syringae. Like other members of this family of bacterial metabolites, it is characterised by a fatty acylated cyclic peptide with mixed chirality and lactonic closure. Several biological activities of Pseudomycin A are lower than those found for some of its congeners, a difference which might depend on the diverse number and distribution of charged residues in the peptide moiety. Hence, it was of interest to investigate its conformation in solution. After the complete interpretation of the two-dimensional NMR spectra, NOE data were obtained and the structure was determined by computer simulations, applying distance geometry and molecular dynamics procedures. The conformation of the large ring of Pseudomycin A in solution includes three rigid structural regions interrupted by three short flexible regions that act as hinges. The overall three-dimensional structure of the cyclic moiety is similar to that of previously studied bioactive lipodepsinonapeptides produced by other pseudomonads.     

MR imaging of focal liver lesions: comparison between turbo spin-echo and conventional spin-echo pulse sequences

The aim of this study was to compare conventional spin-echo (CSE)T ₂-weighted (T2W) images with turbo spin-echo (TSE) T2W pulse sequences in their ability to detect focal liver lesions. Seventy-eight consecutive patients with focal liver lesions were entered into this study. All patients were imaged using the gradient-echo (GE) sequence with the breath-hold technique forT ₁-weighted (T1W) images, and CSE and TSE sequences for T2W images. Qualitative evaluation included lesion detection (number of lesions detected) and conspicuity (extent of visualization of lesional borders); quantitative evaluation included the signal-to-noise (S/N) ratio and the contrast-to-noise (C/N) ratio. TSE showed the best performance in terms of lesion detection; however, the difference between TSE and CSE was significant only in the case of benign cysts (p<0.01). Conspicuity was higher with TSE and CSE, and lower with GE. The S/N and C/N ratios of the two T2W sequences were also comparable, and better than those of GE. However, the combined use of GE and TSE resulted in improved lesion detection. The results show that, because the acquisition time is greatly reduced with TSE sequences, these should be considered as first-line approach to magnetic resonance imaging of the liver for the study of focal lesions.     

Preparation and characterization of comb-shaped polyesters from 2,2-dioctadecyl-1,3-propanediol and phthalic acids

Summary New types of comb-shaped polyesters have been prepared by transesterification of 2,2-dioctadecyl-1,3-propanediol and the three isomeric diphenyl phthalates. Intrinsic viscosity and SEC analyses allowed determination of DPs (60–89) and molar masses; results compare well with absolute average weight molar masses (56,000–118,000) determined by low-angle laser light scattering. High-resolution ¹³C NMR reveals the characteristic aromatic and glycol polyester patterns; it resolves many side-chain methylene carbons, but shows no evidence of end-groups. All the polyesters are crystalline as shown by X-ray diffractometry and DSC, the crystallinity being exclusively due to the octadecyl side-group crystallization in the hexagonal form. The results indicate that the crystallinity decreases substantially when going from the ortho- to the terephthalate polyester.     

Solution structure of glutathione bound to human glutathione transferase P1-1: comparison of NMR measurements with the crystal structure

The conformation of the bound glutathione (GSH) in the active site of the human glutathione transferase P1-1 (EC 2.5.1.18) has been studied by transferred NOE measurements and compared with those obtained by X-ray diffraction data. Two-dimensional TRNOESY and TRROESY experiments have been performed under fast-exchange conditions. The family of GSH conformers, compatible with TRNOE distance constraints, shows a backbone structure very similar to the crystal model. Interesting differences have been found in the side chain regions. After restrained energy minimization of a representative NMR conformer in the active site, the sulfur atom is not found in hydrogen-bonding distance of the hydroxyl group of Tyr 7. This situation is similar to the one observed in an "atypical" crystal complex grown at low pH and low temperature. The NMR conformers display also a poorly defined structure of the glutamyl moiety, and the presence of an unexpected intermolecular NOE could indicate a different interaction of this substrate portion with the G-site. The NMR data seem to provide a snapshot of GSH in a precomplex where the GSH glutamyl end is bound in a different fashion. The existence of this precomplex is supported by pre-steady-state kinetic experiments [Caccuri, A. M., Lo Bello, M., Nuccetelli, M., Nicotra, M., Rossi, P., Antonini, G., Federici, G., and Ricci, G. (1998) Biochemistry 37, 3028-3034] and preliminary time-resolved fluorescence data.     

Indoor air quality in Portuguese schools: levels and sources of pollutants

Indoor air quality (IAQ) parameters in 73 primary classrooms in Porto were examined for the purpose of assessing levels of volatile organic compounds (VOCs), aldehydes, particulate matter, ventilation rates and bioaerosols within and between schools, and potential sources. Levels of VOCs, aldehydes, PM_2.5, PM₁₀, bacteria and fungi, carbon dioxide (CO₂), carbon monoxide, temperature and relative humidity were measured indoors and outdoors and a walkthrough survey was performed concurrently. Ventilation rates were derived from CO₂ and occupancy data. Concentrations of CO₂ exceeding 1000 ppm were often encountered, indicating poor ventilation. Most VOCs had low concentrations (median of individual species <5 μg/m³) and were below the respective WHO guidelines. Concentrations of particulate matter and culturable bacteria were frequently higher than guidelines/reference values. The variability of VOCs, aldehydes, bioaerosol concentrations, and CO₂ levels between schools exceeded the variability within schools. These findings indicate that IAQ problems may persist in classrooms where pollutant sources exist and classrooms are poorly ventilated; source control strategies (related to building location, occupant behavior, maintenance/cleaning activities) are deemed to be the most reliable for the prevention of adverse health consequences in children in schools.     

Effect of stocking density and group size on growth performance,carcass traits and meat quality of outdoor-reared rabbits

The effect of stocking density (16 rabbits/m², 5 rabbits/m², 2.5 rabbits/m², n = 60, Experiment 1) and group size (4 rabbits/cage, 8 rabbits/cage, 16 rabbits/cage, n = 88, Experiment 2) on productive performance, carcass and meat quality of a slow-growing rabbit population reared outdoors was investigated in two experiments. The highest stocking density induced the highest skin percentage. Lower stocking densities showed lower lightness of Biceps femoris and higher redness of Longissimus lumborum muscles. Four rabbits/cage group (Experiment 2) showed the highest daily weight gain and slaughter weight and the lowest skin percentage. The muscles of 16 rabbits/cage showed significantly higher pHu than 8 and 4 rabbits/cage. BF of 16 and 4 rabbits/cage showed higher L* value. Productive performance and meat quality of rabbits reared outdoors improved in low group size while stocking density needs more experiments. The best combination of density, group size and total available surface that showed the best production and carcass traits was of 5 rabbits/m², 4 rabbits/cage, and 0.8 m².     

A Comparison of Network-Based Methods for Drug Repurposing along with an Application to Human Complex Diseases

Drug repurposing strategy, proposing a therapeutic switching of already approved drugs with known medical indications to new therapeutic purposes, has been considered as an efficient approach to unveil novel drug candidates with new pharmacological activities, significantly reducing the cost and shortening the time of de novo drug discovery. Meaningful computational approaches for drug repurposing exploit the principles of the emerging field of Network Medicine, according to which human diseases can be interpreted as local perturbations of the human interactome network, where the molecular determinants of each disease (disease genes) are not randomly scattered, but co-localized in highly interconnected subnetworks (disease modules), whose perturbation is linked to the pathophenotype manifestation. By interpreting drug effects as local perturbations of the interactome, for a drug to be on-target effective against a specific disease or to cause off-target adverse effects, its targets should be in the nearby of disease-associated genes. Here, we used the network-based proximity measure to compute the distance between the drug module and the disease module in the human interactome by exploiting five different metrics (minimum, maximum, mean, median, mode), with the aim to compare different frameworks for highlighting putative repurposable drugs to treat complex human diseases, including malignant breast and prostate neoplasms, schizophrenia, and liver cirrhosis. Whilst the standard metric (that is the minimum) for the network-based proximity remained a valid tool for efficiently screening off-label drugs, we observed that the other implemented metrics specifically predicted further interesting drug candidates worthy of investigation for yielding a potentially significant clinical benefit.     

New Insights in the Control of Fat Homeostasis: The Role of Neurotensin

Neurotensin (NT) is a small peptide with pleiotropic functions, exerting its primary actions by controlling food intake and energy balance. The first evidence of an involvement of NT in metabolism came from studies on the central nervous system and brain circuits, where NT acts as a neurotransmitter, producing different effects in relation to the specific region involved. Moreover, newer interesting chapters on peripheral NT and metabolism have emerged since the first studies on the NT-mediated regulation of gut lipid absorption and fat homeostasis. Intriguingly, NT enhances fat absorption from the gut lumen in the presence of food with a high fat content, and this action may explain the strong association between high circulating levels of pro-NT, the NT stable precursor, and the increased incidence of metabolic disorders, cardiovascular diseases, and cancer observed in large population studies. This review aims to provide a synthetic overview of the main regulatory effects of NT on several biological pathways, particularly those involving energy balance, and will focus on new evidence on the role of NT in controlling fat homeostasis, thus influencing the risk of unfavorable cardio–metabolic outcomes and overall mortality in humans.     

Deep Resequencing of 9 Candidate Genes Identifies a Role for ARAP1 and IGF2BP2 in Modulating Insulin Secretion Adjusted for Insulin Resistance in Obese Southern Europeans

Type 2 diabetes is characterized by impairment in insulin secretion, with an established genetic contribution. We aimed to evaluate common and low-frequency (1–5%) variants in nine genes strongly associated with insulin secretion by targeted sequencing in subjects selected from the extremes of insulin release measured by the disposition index. Collapsing data by gene and/or function, the association between disposition index and nonsense variants were significant, also after adjustment for confounding factors (OR = 0.25, 95% CI = 0.11–0.59, p = 0.001). Evaluating variants individually, three novel variants in ARAP1, IGF2BP2 and GCK, out of eight reaching significance singularly, remained associated after adjustment. Constructing a genetic risk model combining the effects of the three variants, only carriers of the ARAP1 and IGF2BP2 variants were significantly associated with a reduced probability to be in the lower, worst, extreme of insulin secretion (OR = 0.223, 95% CI = 0.105–0.473, p < 0.001). Observing a high number of normal glucose tolerance between carriers, a regression posthoc analysis was performed. Carriers of genetic risk model variants had higher probability to be normoglycemic, also after adjustment (OR = 2.411, 95% CI = 1.136–5.116, p = 0.022). Thus, in our southern European cohort, nonsense variants in all nine candidate genes showed association with better insulin secretion adjusted for insulin resistance, and we established the role of ARAP1 and IGF2BP2 in modulating insulin secretion.