Novel (Phenothiazinyl)Vinyl-Pyridinium Dyes and Their Potential Applications as Cellular Staining Agents

We report here the synthesis and structural characterization of novel cationic (phenothiazinyl)vinyl-pyridinium (PVP) dyes, together with optical (absorption/emission) properties and their potential applicability as fluorescent labels. Convective heating, ultrasound irradiation and mechanochemical synthesis were considered as alternative synthetic methodologies proficient for overcoming drawbacks such as long reaction time, nonsatisfactory yields or solvent requirements in the synthesis of novel dye (E)-1-(3-chloropropyl)-4-(2-(10-methyl-10H-phenothiazin-3-yl)vinyl)pyridin-1-ium bromide 3d and its N-alkyl-2-methylpyridinium precursor 1c. The trans geometry of the newly synthesized (E)-4-(2-(7-bromo-10-ethyl-10H-phenothiazin-3-yl)vinyl)-1-methylpyridin-1-ium iodide 3b and (E)-1-methyl-4-(2-(10-methyl-10H-phenothiazin-3-yl)vinyl)pyridin-1-ium tetrafluoroborate 3a′ was confirmed by single crystal X-ray diffraction. A negative solvatochromism of the dyes in polar solvents was highlighted by UV-Vis spectroscopy and explanatory insights were supported by molecular modeling which suggested a better stabilization of the lowest unoccupied molecular orbitals (LUMO). The photostability of the dye 3b was investigated by irradiation at 365 nm in different solvents, while the steady-state and time-resolved fluorescence properties of dye 3b and 3a′ in solid state were evaluated under one-photon excitation at 485 nm. The in vitro cytotoxicity of the new PVP dyes on B16-F10 melanoma cells was evaluated by WST-1 assay, while their intracellular localization was assessed by epi-fluorescence conventional microscopy imaging as well as one- and two-photon excited confocal fluorescence lifetime imaging microscopy (FLIM). PVP dyes displayed low cytotoxicity, good internalization inside melanoma cells and intense fluorescence emission inside the B16-F10 murine melanoma cells, making them suitable staining agents for imaging applications.     

Chronically Elevated Exogenous Glucose Elicits Antipodal Effects on the Proteome Signature of Differentiating Human iPSC-Derived Pancreatic Progenitors

The past decade revealed that cell identity changes, such as dedifferentiation or transdifferentiation, accompany the insulin-producing β-cell decay in most diabetes conditions. Mapping and controlling the mechanisms governing these processes is, thus, extremely valuable for managing the disease progression. Extracellular glucose is known to influence cell identity by impacting the redox balance. Here, we use global proteomics and pathway analysis to map the response of differentiating human pancreatic progenitors to chronically increased in vitro glucose levels. We show that exogenous high glucose levels impact different protein subsets in a concentration-dependent manner. In contrast, regardless of concentration, glucose elicits an antipodal effect on the proteome landscape, inducing both beneficial and detrimental changes in regard to achieving the desired islet cell fingerprint. Furthermore, we identified that only a subgroup of these effects and pathways are regulated by changes in redox balance. Our study highlights a complex effect of exogenous glucose on differentiating pancreas progenitors characterized by a distinct proteome signature.     

Quantum-inspired design of resilient substitution boxes: From coding to hardware implementation

S-boxes constitute a cornerstone component in symmetric-key cryptographic algorithms, such as DES and AES encryption systems. In block ciphers, they are typically used to obscure the relationship between the plaintext and the ciphertext. Non-linear and non-correlated S-boxes are the most secure against linear and differential cryptanalysis. In this paper, we focus on a twofold objective: first, we evolve regular S-boxes with high non-linearity and low auto-correlation properties; then automatically generate evolvable hardware for the obtained S-box. Targeting the former, we use a quantum-inspired evolutionary algorithm to optimize regularity, non-linearity and auto-correlation, which constitute the three main desired properties in resilient S-boxes. Pursuing the latter, we exploit the same algorithm to automatically generate the evolvable hardware designs of substitution boxes that minimize hardware space and encryption/decryption time, which form the two main hardware characteristics. We compare our results against existing and well-known designs, which were produced by using conventional methods as well as through genetic algorithm. We will show that our approach provides higher quality S-boxes coding as well as circuits.     

TimeLapseAnalyzer: multi-target analysis for live-cell imaging and time-lapse microscopy

The direct observation of cells over time using time-lapse microscopy can provide deep insights into many important biological processes. Reliable analyses of motility, proliferation, invasive potential or mortality of cells are essential to many studies involving live cell imaging and can aid in biomarker discovery and diagnostic decisions. Given the vast amount of image- and time-series data produced by modern microscopes, automated analysis is a key feature to capitalize the potential of time-lapse imaging devices. To provide fast and reproducible analyses of multiple aspects of cell behaviour, we developed TimeLapseAnalyzer. Apart from general purpose image enhancements and segmentation procedures, this extensible, self-contained, modular cross-platform package provides dedicated modalities for fast and reliable analysis of multi-target cell tracking, scratch wound healing analysis, cell counting and tube formation analysis in high throughput screening of live-cell experiments. TimeLapseAnalyzer is freely available (MATLAB, Open Source) at http://www.informatik.uni-ulm.de/ni/mitarbeiter/HKestler/tla.     

Polarized light imaging of white matter architecture

Polarized light imaging (PLI) is a method to image fiber orientation in gross histological brain sections based on the birefringent properties of the myelin sheaths. The method uses the transmission of polarized light to quantitatively estimate the fiber orientation and inclination angles at every point of the imaged section. Multiple sections can be assembled into a 3D volume, from which the 3D extent of fiber tracts can be extracted. This article describes the physical principles of PLI and describes two major applications of the method: the imaging of white matter orientation of the rat brain and the generation of fiber orientation maps of the human brain in white and gray matter. The strengths and weaknesses of the method are set out.     

Compact yet efficient hardware implementation of artificial neural networks with customized topology

There are several neural network implementations using either software, hardware-based or a hardware/software co-design. This work proposes a hardware architecture to implement an artificial neural network (ANN), whose topology is the multilayer perceptron (MLP). In this paper, we explore the parallelism of neural networks and allow on-the-fly changes of the number of inputs, number of layers and number of neurons per layer of the net. This reconfigurability characteristic permits that any application of ANNs may be implemented using the proposed hardware. In order to reduce the processing time that is spent in arithmetic computation, a real number is represented using a fraction of integers. In this way, the arithmetic is limited to integer operations, performed by fast combinational circuits. A simple state machine is required to control sums and products of fractions. Sigmoid is used as the activation function in the proposed implementation. It is approximated by polynomials, whose underlying computation requires only sums and products. A theorem is introduced and proven so as to cover the arithmetic strategy of the computation of the activation function. Thus, the arithmetic circuitry used to implement the neuron weighted sum is reused for computing the sigmoid. This resource sharing decreased drastically the total area of the system. After modeling and simulation for functionality validation, the proposed architecture synthesized using reconfigurable hardware. The results are promising.     

Preference-based multi-objective evolutionary algorithms for power-aware application mapping on NoC platforms

Network-on-chip (NoC) are considered the next generation of communication infrastructure in embedded systems. In the platform-based design methodology, an application is implemented by a set of collaborative intellectual property (IP) blocks. The selection of the most suited set of IPs as well as their physical mapping onto the NoC infrastructure to implement efficiently the application at hand are two hard combinatorial problems that occur during the synthesis process of Noc-based embedded system implementation. In this paper, we propose an innovative preference-based multi-objective evolutionary methodology to perform the assignment and mapping stages. We use one of the well-known and efficient multi-objective evolutionary algorithms NSGA-II and microGA as a kernel. The optimization processes of assignment and mapping are both driven by the minimization of the required silicon area and imposed execution time of the application, considering that the decision maker’s preference is a pre-specified value of the overall power consumption of the implementation.     

Pleurotusostreatus and Agaricus subrufescens: investigation of chemical composition and antioxidant properties of these mushrooms cultivated with different handmade and commercial supplements

In this study, the chemical composition, total phenolic content and antioxidant activity (DPPH, ORAC and FRAP assays) of A. subrufescens and P. ostreatus, cultivated with handmade and commercials supplements, were compared. Additionally, the compounds ergosterol, saccharopine, and hexitol were identified in A. subrufescens by HPLC-MS/MS. The antioxidant compound p-coumaric acid and dihexoses was found in both mushroom species. A. subrufescens presented higher total phenolic content (73.8 ± 0.6 mg GAE 100 g⁻¹) and antioxidant activity than P. ostreatus (16.6 ± 0.5 mg GAE 100 g⁻¹). The handmade supplement based on the waste of noble grains presented statistically similar phenolic content to the mushrooms cultivated with commercial ones Spawn Mate II SE (86.1 ± 1.4 and 92.9 ± 0.3 mg GAE 100 g⁻¹, respectively). Therefore, the results support the use of handmade supplements based on agro-wastes as a viable alternative to the use of high-cost commercial ones.     

Production of fungal lipases using wheat bran and soybean bran and incorporation of sugarcane bagasse as a co-substrate in solid-state fermentation

Fungal strains were screened for lipase producing activities and 10 strains were classified as good producers. Aspergillus sp., Fusarium sp., and Penicillium sp. exhibited the highest activities when fermented in wheat bran (WB) and soybean bran (SB). No fungal growth was observed using sugarcane bagasse (CB). An experimental design was applied to incorporate CB into the fermentation process for lipase production by Aspergillus sp. and Penicillium sp., and to evaluate the best moisture content for the substrate. Strains studied achieved maximum lipase activities with 25% CB combined with 75% WB or SB at 40% moisture content. The highest lipase activities were observed for WB and SB, and for SB combined with CB using Aspergillus sp. Fermentation of 96 h was the optimum period for enzyme production.