Structure,Biosynthesis, and Biological Activity of Succinylated Forms of Bacteriocin BacSp222

BacSp222 is a multifunctional peptide produced by Staphylococcus pseudintermedius 222. This 50-amino acid long peptide belongs to subclass IId of bacteriocins and forms a four-helix bundle molecule. In addition to bactericidal functions, BacSp222 possesses also features of a virulence factor, manifested in immunomodulatory and cytotoxic activities toward eukaryotic cells. In the present study, we demonstrate that BacSp222 is produced in several post-translationally modified forms, succinylated at the ε-amino group of lysine residues. Such modifications have not been previously described for any bacteriocins. NMR and circular dichroism spectroscopy studies have shown that the modifications do not alter the spatial structure of the peptide. At the same time, succinylation significantly diminishes its bactericidal and cytotoxic potential. We demonstrate that the modification of the bacteriocin is an effect of non-enzymatic reaction with a highly reactive intracellular metabolite, i.e., succinyl-coenzyme A. The production of succinylated forms of the bacteriocin depends on environmental factors and on the access of bacteria to nutrients. Our study indicates that the production of succinylated forms of bacteriocin occurs in response to the changing environment, protects producer cells against the autotoxicity of the excreted peptide, and limits the pathogenicity of the strain.     

Evaluation of Polymeric Matrix Loaded with Melatonin for Wound Dressing

The development of scaffolds mimicking the extracellular matrix containing bioactive substances has great potential in tissue engineering and wound healing applications. This study investigates melatonin—a methoxyindole present in almost all biological systems. Melatonin is a bioregulator in terms of its potential clinical importance for future therapies of cutaneous diseases. Mammalian skin is not only a prominent melatonin target, but also produces and rapidly metabolizes the multifunctional methoxyindole to biologically active metabolites. In our methodology, chitosan/collagen (CTS/Coll)-contained biomaterials are blended with melatonin at different doses to fabricate biomimetic hybrid scaffolds. We use rat tail tendon- and Salmo salar fish skin-derived collagens to assess biophysical and cellular properties by (i) Fourier transform infrared spectroscopy—attenuated total reflectance (FTIR–ATR), (ii) thermogravimetric analysis (TG), (iii) scanning electron microscope (SEM), and (iv) proliferation ratio of cutaneous cells in vitro. Our results indicate that melatonin itself does not negatively affect biophysical properties of melatonin-immobilized hybrid scaffolds, but it induces a pronounced elevation of cell viability within human epidermal keratinocytes (NHEK), dermal fibroblasts (NHDF), and reference melanoma cells. These results demonstrate that this indoleamine accelerates re-epithelialization. This delivery is a promising technique for additional explorations in future dermatotherapy and protective skin medicine.     

Recent operational experiences of FILMTEC NF270 membrane in Europe

Nanofiltration (NF) is an attractive technology for potable and industrial water treatment because NF operates between ultrafiltration (UF) and reverse osmosis (RO) membranes. NF is designed to remove a high percentage of organic contaminants (humic acids, pesticides, color bodies) while passing a medium to high percentage of salt. Compared to UF membranes, the NF product water quality is significantly better; compared to comparable RO treatment systems NF systems require lower operating pressures. Due to these features, NF is increasingly used in a broad range of water treatment applications. The general applications include softening, as well as color, organics and micro-organism removal. DOW FILMTEC NF270-400 is one the most frequently used elements in water treatment and this paper presents examples of three recent NF270-400 installations in Europe. The first two plants, Eupen and Stembert, are located in Belgium and produce potable water from surface water. The third one is a Scandinavian plant which purifies groundwater for a brewery and soft drink production. The presented operation results prove NF to be a highly competitive technique for low cost water treatment.     

Peptidomic analysis of rat urine using capillary electrophoresis coupled to mass spectrometry

We have established and validated a protocol for the peptidomic analysis of rat urine using CE coupled to MS (CE-MS). In the first experiments, the reproducibility of the CE-MS set-up and of the established preparation procedure were assessed. To establish a first rat urinary peptidome map, samples were also analyzed using CE-FT-ICR. The subsequent analysis of independent samples from two different strains (WISTAR and CD) indicated strain-specific differences, which were validated in a blinded assessment. MS/MS revealed the presence of specific fragments from well-known urinary rat peptides, such as collagens, alpha-1-antitrypsin, and serum albumin. The CE-MS-based peptidomics platform may provide novel insights into body fluids of animal models, such as rat or mice. Together with peptide identification, the technology appears to be an excellent, complimentary, and non-invasive tool to analyze toxicological or other (patho)physiological effects of pharmaceutical compounds in animal models.     

A computational model of time for stiff hybrid systems applied to control synthesis

Computation has quickly become of paramount importance in the design of engineered systems, both to support their features as well as their design. Tool support for high-level modeling formalisms has endowed design specifications with executable semantics. Such specifications typically include not only discrete-time and discrete-event behavior, but also continuous-time behavior that is stiff from a numerical integration perspective. The resulting stiff hybrid dynamic systems necessitate variable-step solvers to simulate the continuous-time behavior as well as solver algorithms for the simulation of discrete-time and discrete-event behavior. The combined solvers rely on complex computer code which makes it difficult to directly solve design tasks with the executable specifications. To further leverage the executable specifications in design, this work aims to formalize the semantics of stiff hybrid dynamic systems at a declarative level by removing implementation detail and only retaining ‘what’ the computer code does and not ‘how’ it does it. A stream-based approach is adopted to formalize variable-step solver semantics and to establish a computational model of time that supports discrete-time and discrete-event behavior. The corresponding declarative formalization is amenable to computational methods and it is shown how model checking can automatically generate, or synthesize, a feedforward control strategy for a stiff hybrid dynamic system. Specifically, a stamper in a surface mount device is controlled to maintain a low acceleration of the stamped component for a prescribed minimum duration of time.     

Neuromyelitis Optica Spectrum Disorder Treatment—Current and Future Prospects

Neuromyelitis optica (NMO) is an immune-mediated demyelinative disorder of the central nervous system affecting mainly the optical nerves and the spinal cord. The recurrent course of the disease, with exacerbations and incomplete remissions, causes accumulating disability, which has a profound impact upon patients’ quality of life. The discovery of antibodies against aquaporin 4 (AQP4) and their leading role in NMO etiology and the formulation of diagnostic criteria have improved appropriate recognition of the disease. In recent years, there has been rapid progress in understanding the background of NMO, leading to an increasing range of treatment options. On the basis of a review of the relevant literature, the authors present currently available therapeutic strategies for NMO as well as ongoing research in this field, with reference to key points of immune-mediated processes involved in the background of the disease.     

Ancient Bacterial Class Alphaproteobacteria Cytochrome P450 Monooxygenases Can Be Found in Other Bacterial Species

Cytochrome P450 monooxygenases (CYPs/P450s), heme-thiolate proteins, are well-known players in the generation of chemicals valuable to humans and as a drug target against pathogens. Understanding the evolution of P450s in a bacterial population is gaining momentum. In this study, we report comprehensive analysis of P450s in the ancient group of the bacterial class Alphaproteobacteria. Genome data mining and annotation of P450s in 599 alphaproteobacterial species belonging to 164 genera revealed the presence of P450s in only 241 species belonging to 82 genera that are grouped into 143 P450 families and 214 P450 subfamilies, including 77 new P450 families. Alphaproteobacterial species have the highest average number of P450s compared to Firmicutes species and cyanobacterial species. The lowest percentage of alphaproteobacterial species P450s (2.4%) was found to be part of secondary metabolite biosynthetic gene clusters (BGCs), compared other bacterial species, indicating that during evolution large numbers of P450s became part of BGCs in other bacterial species. Our study identified that some of the P450 families found in alphaproteobacterial species were passed to other bacterial species. This is the first study to report on the identification of CYP125 P450, cholesterol and cholest-4-en-3-one hydroxylase in alphaproteobacterial species (Phenylobacterium zucineum) and to predict cholesterol side-chain oxidation capability (based on homolog proteins) by P. zucineum.     

Effect of Enzymatic Hydrolysis of Wheat Starch on Amylose‐Lipid Complexes Stability

In the process of enzymatic hydrolysis of wheat starch to glucose, the presence of amylose‐lipid complexes (AML's) decreases swelling and dissolving capacity and the water binding capacity of starch, thus impeding the access of amylolytic enzymes to the starch granules. The aim of the work was to define the relationship between the stability of AML's and the conditions of the enzymatic hydrolysis of wheat starch such as the kind of enzymatic preparation (only amylolytic or both amylolytic and lipolytic) and time of hydrolysis. Hydrolysates were produced from wheat starch liquefied with bacterial α‐amylase BAN 240L, subjected to further treatment with the enzymatic preparation Spezyme GA 300W, containing glucoamylase and lysophospholipase and, for comparison, only with a glucoamylase preparation (AMG 300L). The effect of amylolytic and lipolytic enzymes on the stability of AML's in the process of wheat starch hydrolysis was estimated. The amount of fatty acids released during hydrolysis was determined with gas‐liquid chromatography (GLC) and by differential scanning calorimetry (DSC) measuring the enthalpy of decomposition of AML's. The investigations revealed a differentiated effect of the individual enzymatic preparations on the degree of degradation of AML's. Amylose‐lipid complexes were more susceptible to the attack of preparation Spezyme GA 300W than to the digestion by α‐amylase BAN 240 L and glucoamylase AMG 300L.