Chromogranin A: From Laboratory to Clinical Aspects of Patients with Neuroendocrine Tumors

Background. Neuroendocrine tumors (NETs) are characterized by having behavior and prognosis that depend upon tumor histology, primary site, staging, and proliferative index. The symptoms associated with carcinoid syndrome and vasoactive intestinal peptide tumors are treated with octreotide acetate. The PROMID trial assesses the effect of octreotide LAR on the tumor growth in patients with well-differentiated metastatic midgut NETs. The CLARINET trial evaluates the effects of lanreotide in patients with nonfunctional, well-, or moderately differentiated metastatic enteropancreatic NETs. Everolimus has been approved for the treatment of advanced pancreatic NETs (pNETs) based on positive PFS effects, obtained in the treated group. Sunitinib is approved for the treatment of patients with progressive gastrointestinal stromal tumor or intolerance to imatinib, because a randomized study demonstrated that it improves PFS and overall survival in patients with advanced well-differentiated pNETs. In a phase II trial, pasireotide shows efficacy and tolerability in the treatment of patients with advanced NETs, whose symptoms of carcinoid syndrome were resistant to octreotide LAR. An open-label, phase II trial assesses the clinical activity of long-acting repeatable pasireotide in treatment-naive patients with metastatic grade 1 or 2 NETs. Even if the growth of the neoplasm was significantly inhibited, it is still unclear whether its antiproliferative action is greater than that of octreotide and lanreotide. Because new therapeutic options are needed to counter the natural behavior of neuroendocrine tumors, it would also be useful to have a biochemical marker that can be addressed better in the management of these patients. Chromogranin A is currently the most useful biomarker to establish diagnosis and has some utility in predicting disease recurrence, outcome, and efficacy of therapy

Genetic variability of Nero Lucano pig breed at IGF2, LEP, MC4R, PIK3C3, RYR1 and VRTN loci

The Nero Lucano pig is a native breed of Southern Italy which thanks to the joint action of Basilicata Region Institutions, University of Basilicata and breeders returned to populate the area of origin. In order to characterise and to monitor the variability present in the population, we genotyped 229 animals at 12 polymorphic loci located in the following genes: IGF2, LEP, MC4R, PIK3C3, RYR1 and VRTN. According to the results three loci (IGF2 209G>C, PIK3C3 2058A>G and RYR1 1843C>T) did not show variability, while the others showed genotype distributions in agreement with Hardy-Weinberg equilibrium and a minor allele frequency ranging from 0.022 for MC4R 892A to 0.479 for PIK3C3 2604T alleles. The IGF2, MC4R and VRTN loci were characterised by very low frequencies (from 0.02 to 0.05) of the alleles that are associated with favourable productive characteristics in cosmopolitan breeds.Highlights Analyses of the genetic variability of Nero Lucano pig population useful for meat production selection plans. The IGF2, MC4R and VRTN loci of Nero Lucano pig show very low frequencies of alleles associated with positive effects on meat production. The Nero Lucano pig can be considered as free from Malignant Hyperthermia, a positive result for the quality of cured meat products

Increased levels of DNA methyltransferases are associated with the tumorigenic capacity of prostate cancer cells

DNA methylation might be the earliest somatic genome changes in prostate cancer that also play an important role in the process of tumor invasion, growth and metastasis. In recent years, several inhibitors of DNA methyltransferases (DNMTis) have been developed and evaluated in pre-clinical models and in clinical trials. While these compounds are effective in the treatment of hematological conditions, clinical trials in solid tumors and in prostate cancer have shown limited or no efficacy. This may be attributed to inappropriate dose regimens leading to toxicity-related adverse events. As with other anti-target compounds, one of the obstacles encountered with DNMTis in prostate cancer could be the inability to select patients for the clinical studies as well as the inability to monitor the efficacy of the drug if not the conclusion of the study. Primary cultures derived from human prostatic tissues harvested from patients with benign prostatic hyperplasia (BPH) and prostate cancer (PCa) as well as neoplastic and non-neoplastic prostate cell lines were tested for DNMT expression/activity and to monitor azacitidine molecular efficacy. We observed that in primary cultures the levels of DNMT activity as well as the protein levels of DNMT1, DNMT3a and DNMT3b were higher in cultures derived from PCa compared to BPH tissue samples and significantly higher in cultures derived from PCa with Gleason scores ≥7 compared to those observed in cultures derived from Gleason scores <7. In addition, DNMT activity as well as DNMT1, DNMT3a and DNMT3b levels were higher in PCa cell lines compared to their non-neoplastic counterparts. Although DNMT activity was higher in high tumorigenic/aggressive PCa cell lines compared to low tumorigenic/aggressive cell lines, only the levels of DNMT3a and DNMT3b were significantly higher in the first group of cells, suggesting that DNMT1 activity is related to the transition to non-neoplastic versus neoplastic phenotype whereas the de novo methylation enzymes were mainly related to progression. Nevertheless, the comparison in the more aggressive PC3 cell derivatives (PC3-LN4 cells) also possessed higher levels of DNMT1 compared to PC3 and PC3M from which these cells were derived. Collectively, our results confirm previous data on the increased methylation in more aggressive tumors supporting the use of DNMTis in advanced prostate cancer. In addition, since glutathione S-transferase-π (GSTP1) was re-expressed or its protein levels were increased after treatment with non-toxic azacitidine doses and since GSTP1 can easily be measured in patient sera, the monitoring of this protein may aide in the evaluation of therapy in future clinical trials

Amino-functionalized poly(l-lactide) lamellar single crystals as a valuable substrate for delivery of HPV16-E7 tumor antigen in vaccine development

Background: Poly(L-lactide) (PLLA) is a biodegradable polymer currently used in many biomedical applications, including the production of resorbable surgical devices, porous scaffolds for tissue engineering, nanoparticles and microparticles for the controlled release of drugs or antigens. The surfaces of lamellar PLLA single crystals (PLLAsc) were provided with amino groups by reaction with a multifunctional amine and used to adsorb an Escherichia coli-produced human papillomavirus (HPV)16-E7 protein to evaluate its possible use in antigen delivery for vaccine development. Methods: PLLA single crystals were made to react with tetraethylenepentamine to obtain amino-functionalized PLLA single crystals (APLLAsc). Pristine and amino-functionalized PLLAsc showed a two-dimensional microsized and one-dimensional nanosized lamellar morphology, with a lateral dimension of about 15–20 µm, a thickness of about 12 nm, and a surface specific area of about 130 m2/g. Both particles were characterized and loaded with HPV16-E7 before being administered to C57BL/6 mice for immunogenicity studies. The E7-specific humoral-mediated and cell-mediated immune response as well as tumor protective immunity were analyzed in mice challenged with TC-1 cancer cells. Results: Pristine and amino-functionalized PLLAsc adsorbed similar amounts of E7 protein, but in protein-release experiments E7-PLLAsc released a higher amount of protein than E7-APLLAsc. When the complexes were dried for observation by scanning electron microscopy, both samples showed a compact layer, but E7-APLLAsc showed greater roughness than E7-PLLAsc. Immunization experiments in mice showed that E7-APLLAsc induced a stronger E7-specific immune response when compared with E7-PLLAsc. Immunoglobulin G isotyping and interferon gamma analysis suggested a mixed Th1/Th2 immune response in both E7-PLLAsc-immunized and E7-APLLAsc-immunized mice. However, only the mice receiving E7-APLLAsc were fully protected from TC-1 tumor growth after three doses of vaccine. Conclusion: Our results show that APLLA single crystals improve the immunogenicity of HPV16-E7 and indicate that E7-APLLAsc could be used for development of an HPV16 therapeutic vaccine against HPV16-related tumors

Infrared evidence of a Slater metal-insulator transition in NaOsO3

The magnetically driven metal-insulator transition (MIT) was predicted by Slater in the fifties. Here a long-range antiferromagnetic (AF) order can open up a gap at the Brillouin electronic band boundary regardless of the Coulomb repulsion magnitude. However, while many low-dimensional organic conductors display evidence for an AF driven MIT, in three-dimensional (3D) systems the Slater MIT still remains elusive. We employ terahertz and infrared spectroscopy to investigate the MIT in the NaOsO3 3D antiferromagnet. From the optical conductivity analysis we find evidence for a continuous opening of the energy gap, whose temperature dependence can be well described in terms of a second order phase transition. The comparison between the experimental Drude spectral weight and the one calculated through Local Density Approximation (LDA) shows that electronic correlations play a limited role in the MIT. All the experimental evidence demonstrates that NaOsO3 is the first known 3D Slater insulator.Comment: 4 figure

Stochastic response of a fractional vibroimpact system

The paper proposes a method to investigate the stochastic dynamics of a vibroimpact single-degree-of-freedom fractional system under a Gaussian white noise input. It is assumed that the system has a hard type impact against a one-sided motionless barrier, which is located at the system's equilibrium position; furthermore, the system under study is endowed with an element modeled with fractional derivative. The proposed method is based on stochastic averaging technique and overcome the particular difficulty due to the presence of fractional derivative of an absolute value function; particularly an analytical expression for the system's mean squared response amplitude is presented and compared with results obtained by numerical simulations

Effect of water contamination on the shear bond strength of self-ligating brackets

AbstractPurposeThe aim of the present study is to evaluate the effect of water contamination on the shear bond strength (SBS) and adhesive remnant index (ARI) score of self-ligating brackets.Materials and methodsOne conventional bracket and three different self-ligating brackets were bonded onto 160 bovine permanent mandibular incisors, divided randomly into 8 groups. For each type of bracket, 20 samples were bonded on dry enamel and 20 after water contamination. After 24h, all specimens were tested for SBS using an Instron Universal Testing Machine, and ARI scores were evaluated.ResultsAll groups showed clinically adequate SBSs. Quick brackets bonded onto dry enamel showed significantly higher SBSs than all other groups tested, whereas the lowest shear strength values were recorded for Step, Quick, and Damon 3MX brackets bonded onto contaminated enamel and for Damon 3MX onto dry enamel. Frequency distribution of ARI Scores showed a prevalence of ARI “2” and “3” for all the groups tested.ConclusionsWater contamination reduces the SBS of self-ligating brackets, but significant differences have been found only for Quick brackets. All groups showed a significant higher frequency of ARI Score of “2” and “3”

Oxidative Status of Goats with Different CSN1S1 Genotypes Fed ad Libitum with Fresh and Dry Forages

Forty late-lactation Girgentana goats were used to study the effect of diets fed ad libitum and αS1-casein (CSN1S1) genotype on redox balance. The goats genotyped at CSN1S1 locus (A/A, A/F) were subjected to four feeding treatments different for percentage inclusion of dry and fresh forage: DAF100 (98% of Dry Alfalfa Forage), DAF65 (65% of Dry Alfalfa Forage), FSF100 (100% of Fresh Sulla Forage) and FSF65 (65% of Fresh Sulla Forage). Blood samples were analyzed for superoxide dismutase (SOD) and glutathione peroxidase (GPX) activity, reactive oxygen metabolites (ROMs), biological antioxidant potential (BAP) and non-esterified fatty acids (NEFA), beta-hydroxybutyrate (BHBA), albumin, glucose and cholesterol contents. The oxidative stress index (OSI) was calculated as percentage ratio of ROMs to BAP. Redox balance was improved by Sulla inclusion, as reflected in the lower OSI values found in FSF100 and FSF65 groups. DAF100 group displayed the highest GPX activity, while other groups exhibited the highest SOD activity. Fresh forage diets increased albumin concentration while no effect of tested factors was noted on glucose, NEFA, BHBA and cholesterol contents. The interaction diet × genotype was significant only for GPX activity. GPX and albumin were negatively correlated and were correlated positively and negatively with ROMs, respectively. Diet rather than genotype affects redox balance in dairy goats and a possible role of forage polyphenol compounds on oxidative status needs to be tested in future studies

Mitotic cell death induction by targeting the mitotic spindle with tubulin-inhibitory indole derivative molecules

Tubulin-targeting molecules are widely used cancer therapeutic agents. They inhibit microtubule-based structures, including the mitotic spindle, ultimately preventing cell division. The final fates of microtubule-inhibited cells are however often heterogeneous and difficult to predict. While recent work has provided insight into the cell response to inhibitors of microtubule dynamics (taxanes), the cell response to tubulin polymerization inhibitors remains less well characterized. Arylthioindoles (ATIs) are recently developed tubulin inhibitors. We previously identified ATI members that effectively inhibit tubulin polymerization in vitro and cancer cell growth in bulk cell viability assays. Here we characterise in depth the response of cancer cell lines to five selected ATIs. We find that all ATIs arrest mitotic progression, yet subsequently yield distinct cell fate profiles in time-lapse recording assays, indicating that molecules endowed with similar tubulin polymerization inhibitory activity in vitro can in fact display differential efficacy in living cells. Individual ATIs induce cytological phenotypes of increasing severity in terms of damage to the mitotic apparatus. That differentially triggers MCL-1 down-regulation and caspase-3 activation, and underlies the terminal fate of treated cells. Collectively, these results contribute to define the cell response to tubulin inhibitors and pinpoint potentially valuable molecules that can increase the molecular diversity of tubulin-targeting agents

EEG-based mental workload neurometric to evaluate the impact of different traffic and road conditions in real driving settings

Car driving is considered a very complex activity, consisting of different concomitant tasks and subtasks, thus it is crucial to understand the impact of different factors, such as road complexity, traffic, dashboard devices, and external events on the driver’s behavior and performance. For this reason, in particular situations the cognitive demand experienced by the driver could be very high, inducing an excessive experienced mental workload and consequently an increasing of error commission probability. In this regard, it has been demonstrated that human error is the main cause of the 57% of road accidents and a contributing factor in most of them. In this study, 20 young subjects have been involved in a real driving experiment, performed under different traffic conditions (rush hour and not) and along different road types (main and secondary streets). Moreover, during the driving tasks different specific events, in particular a pedestrian crossing the road and a car entering the traffic flow just ahead of the experimental subject, have been acted. A Workload Index based on the Electroencephalographic (EEG), i.e., brain activity, of the drivers has been employed to investigate the impact of the different factors on the driver’s workload. Eye-Tracking (ET) technology and subjective measures have also been employed in order to have a comprehensive overview of the driver’s perceived workload and to investigate the different insights obtainable from the employed methodologies. The employment of such EEG-based Workload index confirmed the significant impact of both traffic and road types on the drivers’ behavior (increasing their workload), with the advantage of being under real settings. Also, it allowed to highlight the increased workload related to external events while driving, in particular with a significant effect during those situations when the traffic was low. Finally, the comparison between methodologies revealed the higher sensitivity of neurophysiological measures with respect to ET and subjective ones. In conclusion, such an EEG-based Workload index would allow to assess objectively the mental workload experienced by the driver, standing out as a powerful tool for research aimed to investigate drivers’ behavior and providing additional and complementary insights with respect to traditional methodologies employed within road safety research