Thermostable Esterase 2 from Alicyclobacillus acidocaldarius as Biosensor for the Detection of Organophosphate Pesticides

Pesticides are the plague of modern times, although much needed in agriculture, causing damage to the entire ecosystem, including humans. The high operative costs and the requirement of specialized personnel for pesticide detection, incentive to develop alternative solutions such as the set up of cheap, rapid, and simple to use biosensors. In this work, we evaluate the possibility to use the esterase 2 from Alicyclobacillus acidocaldarius as a biosensor for the detection of specific organophosphate pesticides. With the recent demonstration of the very high affinity of esterase 2 toward paraoxon, a more complete analysis on the detection methods in water as well as in purposely contaminated fruit juices was carried out. The inhibitory effects of a wide range of other pesticides on esterase 2 were investigated, showing a better selectivity with respect to nonspecific reaction of acethylcholinesterases, the main target of organophosphate pesticides. The applied methodology allowed one to detect 2.75 × 10(-3) ppm of neurotoxic agent, comparable to the efficiency of other acethylcholinesterase-based biosensors. Finally, a raw biosensor, based on EST2 immobilization on a nitrocellulose membrane, was devised and tested for paraoxon detection, showing longtime stability, reproducibility, and sensibility.     

Les phases SrLa2Al2O7 et SrGd2Al2O7

SrLa₂Al₂O₇ and SrGd₂Al₂O₇ belong to the structural type Sr₃Ti₂O₇. In SrGd₂Al₂O₇ strontium and gadolinium occupy respectively the 12 and 9 coordinated sites, whereas in SrLa₂Al₂O₇ the homologous cations are statistically distributed. The fluorescence spectra of both phases activated by Eu³⁺ ions show that the only possible position of Eu³⁺ is the 9 coordinated site.     

State-of-the-Art on Wound Vitality Evaluation: A Systematic Review

The vitality demonstration refers to determining if an injury has been caused ante- or post-mortem, while wound age means to evaluate how long a subject has survived after the infliction of an injury. Histology alone is not enough to prove the vitality of a lesion. Recently, immunohistochemistry, biochemistry, and molecular biology have been introduced in the field of lesions vitality and age demonstration. The study was conducted according to the preferred reporting items for systematic review (PRISMA) protocol. The search terms were “wound”, “lesion”, “vitality”, “evaluation”, “immunohistochemistry”, “proteins”, “electrolytes”, “mRNAs”, and “miRNAs” in the title, abstract, and keywords. This evaluation left 137 scientific papers. This review aimed to collect all the knowledge on vital wound demonstration and provide a temporal distribution of the methods currently available, in order to determine the age of lesions, thus helping forensic pathologists in finding a way through the tangled jungle of wound vitality evaluation.     

The Marching Intersections algorithm for merging range images

A new algorithm for the integration of partially overlapping range images into a triangular mesh is presented. The algorithm consists of three main steps: it locates the intersections between the range surfaces and a reference grid chosen by the user, then merges all nearly coincident and redundant intersections according to a proximity criterion, and, finally, reconstructs the merged surface(s) from the filtered intersection set. Compared with previous methods, which adopt a volumetric approach, our algorithm shows lower computational costs and improves the accuracy of the surfaces produced. It takes into account the quality of the input measurements and is able to patch small holes corresponding to the parts of the 3D scanned object that were not observed by the acquisition device. The algorithm has been tested on several datasets of range maps; graphical and numeric results are reported.     

Petri net based process scheduling: A model of the control system of flexible manufacturing systems

In this paper, we propose a class of algorithms for the sub-optimal solution of a particular class of problems of process scheduling, particularly focusing on a case study in the area of flexible manufacturing systems (FMSs). The general class of problems we face in our approach is characterized as follows: there is a set of concurrent processes, each formed by a number of temporally related tasks (segments). Tasks are executable by alternate resource sets, different both in performance and costs. Processes and tasks are characterized by release times, due dates, and deadlines. Time constraints are also present in the availability of each resource in resource sets. It has been proven that such a problem does not admit an algorithm for an optimal solution in polynomial time. Our proposed algorithm finds a sub-optimal schedule according to a set of optimization criteria, based on task and process times (earliness, tardiness), and/or time independent costs of resources. Our approach to process scheduling is based on Timed Coloured Petri Nets. We describe the structure of the coordination and scheduling algorithms, concentrating on (i) the general-purpose component, and (ii) the application-dependent component. In particular, the paper focuses on the following issues: (i) theautomatic synthesis of Petri net models of the coordination subsystem, starting from the problem knowledge base; (ii) the dynamic behavior of the coordination subsystem, whose kernel is a High Level Petri net executor, a coordination process based on an original, general purpose algorithm; (iii) the structure of the real-time scheduling subsystem, based on particular heuristic sub-optimal multi-criteria algorithms. Furthermore, the paper defines the interaction mechanisms between the coordination and scheduling subsystems. Our approach clearly distinguishes the mechanism of the net execution from the decision support system. Two conceptually distinct levels, which correspond to two different, interacting implementation modules in the prototype CASE tool, have been defined: theexecutor and thescheduler levels. One of the outstanding differences between these levels is that the executor is conceived as a fast, efficient coordination process, without special-purpose problem-solving capabilities in case of conflicts. The scheduler, on the other hand, is the adaptive, distributed component, whose behavior may heavily depend on the problem class. If the scheduler fails, the executor is, in any case, able to proceed with a general-purpose conflict resolution strategy. Experimental results on the real-time performance of the kernel of the implemented system are finally shown in the paper. The approach described in this paper is at the basis of a joint project with industrial partners for the development of a CASE tool for the simulation of blast furnaces.     

Energy Deposition around Swift Carbon-Ion Tracks in Liquid Water

Energetic carbon ions are promising projectiles used for cancer radiotherapy. A thorough knowledge of how the energy of these ions is deposited in biological media (mainly composed of liquid water) is required. This can be attained by means of detailed computer simulations, both macroscopically (relevant for appropriately delivering the dose) and at the nanoscale (important for determining the inflicted radiobiological damage). The energy lost per unit path length (i.e., the so-called stopping power) of carbon ions is here theoretically calculated within the dielectric formalism from the excitation spectrum of liquid water obtained from two complementary approaches (one relying on an optical-data model and the other exclusively on ab initio calculations). In addition, the energy carried at the nanometre scale by the generated secondary electrons around the ion’s path is simulated by means of a detailed Monte Carlo code. For this purpose, we use the ion and electron cross sections calculated by means of state-of-the art approaches suited to take into account the condensed-phase nature of the liquid water target. As a result of these simulations, the radial dose around the ion’s path is obtained, as well as the distributions of clustered events in nanometric volumes similar to the dimensions of DNA convolutions, contributing to the biological damage for carbon ions in a wide energy range, covering from the plateau to the maximum of the Bragg peak.     

Iron Mobilization from Ferritin in Yeast Cell Lysate and Physiological Implications

Most in vitro iron mobilization studies from ferritin have been performed in aqueous buffered solutions using a variety of reducing substances. The kinetics of iron mobilization from ferritin in a medium that resembles the complex milieu of cells could dramatically differ from those in aqueous solutions, and to our knowledge, no such studies have been performed. Here, we have studied the kinetics of iron release from ferritin in fresh yeast cell lysates and examined the effect of cellular metabolites on this process. Our results show that iron release from ferritin in buffer is extremely slow compared to cell lysate under identical experimental conditions, suggesting that certain cellular metabolites present in yeast cell lysate facilitate the reductive release of ferric iron from the ferritin core. Using filtration membranes with different molecular weight cut-offs (3, 10, 30, 50, and 100 kDa), we demonstrate that a cellular component >50 kDa is implicated in the reductive release of iron. When the cell lysate was washed three times with buffer, or when NADPH was omitted from the solution, a dramatic decrease in iron mobilization rates was observed. The addition of physiological concentrations of free flavins, such as FMN, FAD, and riboflavin showed about a two-fold increase in the amount of released iron. Notably, all iron release kinetics occurred while the solution oxygen level was still high. Altogether, our results indicate that in addition to ferritin proteolysis, there exists an auxiliary iron reductive mechanism that involves long-range electron transfer reactions facilitated by the ferritin shell. The physiological implications of such iron reductive mechanisms are discussed.     

Light Triggers the Antiproliferative Activity of Naphthalimide-Conjugated (η6-arene)ruthenium(II) Complexes

We report the synthesis and characterization of three half-sandwich Ru(II) arene complexes [(η⁶-arene)Ru(N,N′)L][PF₆]₂ containing arene = p-cymene, N,N′ = bipyridine, and L = pyridine meta- with methylenenaphthalimide (C1), methylene(nitro)naphthalimide (C2), or methylene(piperidinyl)naphthalimide (C3). The naphthalimide acts as an antenna for photoactivation. After 3 h of irradiation with blue light, the monodentate pyridyl ligand had almost completely dissociated from complex C3, which contains an electron donor on the naphthalimide ring, whereas only 50% dissociation was observed for C1 and C2. This correlates with the lower wavelength and strong absorption of C3 in this region of the spectrum (λ_max = 418 nm) compared with C1 and C2 (λ_max = 324 and 323 nm, respectively). All the complexes were relatively non-toxic towards A549 human lung cancer cells in the dark, but only complex C3 exhibited good photocytoxicity towards these cancer cells upon irradiation with blue light (IC₅₀ = 10.55 ± 0.30 μM). Complex C3 has the potential for use in photoactivated chemotherapy (PACT).     

Potential of the Stromal Matricellular Protein Periostin as a Biomarker to Improve Risk Assessment in Prostate Cancer

Prostate cancer (PCa) ranges from indolent to aggressive tumors that may rapidly progress and metastasize. The switch to aggressive PCa is fostered by reactive stroma infiltrating tumor foci. Therefore, reactive stroma-based biomarkers may potentially improve the early detection of aggressive PCa, ameliorating disease classification. Gene expression profiles of PCa reactive fibroblasts highlighted the up-regulation of genes related to stroma deposition, including periostin and sparc. Here, the potential of periostin as a stromal biomarker has been investigated on PCa prostatectomies by immunohistochemistry. Moreover, circulating levels of periostin and sparc have been assessed in a low-risk PCa patient cohort enrolled in active surveillance (AS) by ELISA. We found that periostin is mainly expressed in the peritumoral stroma of prostatectomies, and its stromal expression correlates with PCa grade and aggressive disease features, such as the cribriform growth. Moreover, stromal periostin staining is associated with a shorter biochemical recurrence-free survival of PCa patients. Interestingly, the integration of periostin and sparc circulating levels into a model based on standard clinico-pathological variables improves its performance in predicting disease reclassification of AS patients. In this study, we provide the first evidence that circulating molecular biomarkers of PCa stroma may refine risk assessment and predict the reclassification of AS patients.