BRAF Gene and Melanoma: Back to the Future

As widely acknowledged, 40–50% of all melanoma patients harbour an activating BRAF mutation (mostly BRAF V600E). The identification of the RAS–RAF–MEK–ERK (MAP kinase) signalling pathway and its targeting has represented a valuable milestone for the advanced and, more recently, for the completely resected stage III and IV melanoma therapy management. However, despite progress in BRAF-mutant melanoma treatment, the two different approaches approved so far for metastatic disease, immunotherapy and BRAF+MEK inhibitors, allow a 5-year survival of no more than 60%, and most patients relapse during treatment due to acquired mechanisms of resistance. Deep insight into BRAF gene biology is fundamental to describe the acquired resistance mechanisms (primary and secondary) and to understand the molecular pathways that are now being investigated in preclinical and clinical studies with the aim of improving outcomes in BRAF-mutant patients.     

Peptides to Tackle Leishmaniasis: Current Status and Future Directions

Peptide-based drugs are an attractive class of therapeutic agents, recently recognized by the pharmaceutical industry. These molecules are currently being used in the development of innovative therapies for diverse health conditions, including tropical diseases such as leishmaniasis. Despite its socioeconomic influence on public health, leishmaniasis remains long-neglected and categorized as a poverty-related disease, with limited treatment options. Peptides with antileishmanial effects encountered to date are a structurally heterogeneous group, which can be found in different natural sources—amphibians, reptiles, insects, bacteria, marine organisms, mammals, plants, and others—or inspired by natural toxins or proteins. This review details the biochemical and structural characteristics of over one hundred peptides and their potential use as molecular frameworks for the design of antileishmanial drug leads. Additionally, we detail the main chemical modifications or substitutions of amino acid residues carried out in the peptide sequence, and their implications in the development of antileishmanial candidates for clinical trials. Our bibliographic research highlights that the action of leishmanicidal peptides has been evaluated mainly using in vitro assays, with a special emphasis on the promastigote stage. In light of these findings, and considering the advances in the successful application of peptides in leishmaniasis chemotherapy, possible approaches and future directions are discussed here.     

PGRP-LB: An Inside View into the Mechanism of the Amidase Reaction

Peptidoglycan recognition proteins (PGRPs) are ubiquitous among animals and play pivotal functions in insect immunity. Non-catalytic PGRPs are involved in the activation of immune pathways by binding to the peptidoglycan (PGN), whereas amidase PGRPs are capable of cleaving the PGN into non-immunogenic compounds. Drosophila PGRP-LB belongs to the amidase PGRPs and downregulates the immune deficiency (IMD) pathway by cleaving meso-2,6-diaminopimelic (meso-DAP or DAP)-type PGN. While the recognition process is well analyzed for the non-catalytic PGRPs, little is known about the enzymatic mechanism for the amidase PGRPs, despite their essential function in immune homeostasis. Here, we analyzed the specific activity of different isoforms of Drosophila PGRP-LB towards various PGN substrates to understand their specificity and role in Drosophila immunity. We show that these isoforms have similar activity towards the different compounds. To analyze the mechanism of the amidase activity, we performed site directed mutagenesis and solved the X-ray structures of wild-type Drosophila PGRP-LB and its mutants, with one of these structures presenting a protein complexed with the tracheal cytotoxin (TCT), a muropeptide derived from the PGN. Only the Y78F mutation abolished the PGN cleavage while other mutations reduced the activity solely. Together, our findings suggest the dynamic role of the residue Y78 in the amidase mechanism by nucleophilic attack through a water molecule to the carbonyl group of the amide function destabilized by Zn²⁺.     

“Modification V” of poly 4-methylpentene-1 from cyclopentane solutions and gels

A new crystalline structure of poly 4-methylpentene-1 (P4MP1), modiification named modification V, is obtained from cyclopentane solutions and gels, for polymer volume fractions between 0.01 and 0.10. The effect of the thermal history imparted to the solution is analyzed. The relation between gelation, polymorphism and existence of helical conformations of P4MP1 in solution is discussed. Modification V is tentatively indexed on the basis of an hexagonal unit cell with dimensions a = 22.17 ± 0.14 Å and c = 6.69 ±0.02 Å. The crystal transforms into modification I at 130 ± 5°C, the heat of transition being + 15 ±2 J.g^?1.     

SEM study of the growth of electrochemically obtained polyselenophene thin films. Effect of electrolyte and monomer concentration in acetonitrile

Summary The influence of electrolyte and monomer concentration on properties and morphology of electrochemically obtained polyselenophene films doped and undoped with hexafluorophosphate and tetrafluoroborate anions was analyzed. The films were deposited on SnO₂-coated glass in a classical three-electrode cell from the monomer and the electrolyte in anhydrous acetonitrile.The electrochemical growth of polyselenophene thin films was studied by X-ray photoelectron spectroscopy (XPS), electron probe microanalysis (EPMA), and scanning electron microscopy (SEM). The morphology study revealed different three- dimensional (3D) and bidimensional (2D) growth, depending on monomer and electrolyte concentration.     

Zinc Coated Steel/Epoxy Adhesive Systems: Investigation of the Interfacial Zone by FTIR Spectroscopy

The present study takes advantage of the ability of Fourier Transform Infrared Spectroscopy (FTIR) for the analysis of ultrathin organic films on metals. FTIR in the reflection mode (IRRAS) is used in order to study the interaction of ultrathin films of dicyandiamide (hardener of most one-pack epoxy resins) with various substrates, model ones such as gold or zinc and industrial ones such as steel and zinc-coated steels.

Pure zinc surfaces and, to a lesser extent, zinc-coated steels are shown to react with dicyandiamide after heating at 180°C, as evidenced by the frequency shift of the absorption band (at about 2200 cm^-1) characteristic for nitrile groups. As real systems consist of thick layers of a fully formulated adhesive cured onto a metallic substrate, the direct investigation of such a buried interphase is no longer possible by FTIR and by most of the known spectroscopies. Some mechanically tested specimens are then analysed, after failure, by FTIR microspectrometry. The spectra obtained, corresponding to the fracture initiation zone which is about 100 μm in diameter, advocate for the presence of an ultrathin layer of modified polymer still covering the substrate.     

Calibration Model for Water Distribution Network Using Pressures Estimated by Artificial Neural Networks

The success of hydraulic simulation models of water distribution networks is associated with the ability of these models to represent real systems accurately. To achieve this, the calibration phase is essential. Current calibration methods are based on minimizing the error between measured and simulated values of pressure and flow. This minimization is based on a search of parameter values to be calibrated, including pipe roughness, nodal demand, and leakage flow. The resulting hydraulic problem contains several variables. In addition, a limited set of known monitored pressure and flow values creates an indeterminate problem with more variables than equations. Seeking to address the lack of monitored data for the calibration of Water Distribution Networks (WDNs), this paper uses a meta-model based on an Artificial Neural Network (ANN) to estimate pressure on all nodes of a network. The calibration of pipe roughness applies a metaheuristic search method called Particle Swarm Optimization (PSO) to minimize the objective function represented by the difference between simulated and forecasted pressure values. The proposed method is evaluated at steady state and over an extended period for a real District Metering Area (DMA), named Campos do Conde II, and the hypothetical network named C-town, which is used as a benchmark for calibration studies.     

Analytical ray tracing system: Introducing art, a C-library designed for seismic applications

Ray tracing technique is an important tool not only to forward but also for inverse problems in Geophysics, which most of the seismic processing steps depend on. However, implementing ray tracing codes can be very time consuming. This article presents a computer library to trace rays in 2.5D media composed by a stack of layers. The velocity profile inside each layer is such that the eikonal equation can be analytically solved. Therefore, the ray tracing within such profile is made fast and accurate. The great advantage of an analytical ray tracing library is the numerical precision of the quantities computed and the fast execution of the implemented codes. Even though ray tracing programs exist for a long time, for example the seis88 package by ?ervený, most of those programs use a numerical approach to compute the ray. Regardless of the fact that numerical methods can solve more general problems, the analytical ones could be part of a more sophisticated simulation process, where the ray tracing time is completely relevant. We demonstrate the feasibility of our codes using several examples (Miqueles et al., 2013)  [1]. The library can also be used for other applications besides seismic, e.g., optics and tomography.