Gouda Cheese with Modified Content of β-Casein as a Source of Peptides with ACE- and DPP-IV-Inhibiting Bioactivity: A Study Based on In Silico and In Vitro Protocol

In silico and in vitro methods were used to analyze ACE- and DPP-IV-inhibiting potential of Gouda cheese with a modified content of β-casein. Firstly, the BIOPEP-UWM database was used to predict the presence of ACE and DPP-IV inhibitors in casein sequences. Then, the following Gouda cheeses were produced: with decreased, increased, and normative content of β-casein after 1 and 60 days of ripening each (six variants in total). Finally, determination of the ACE/DPP-IV-inhibitory activity and the identification of peptides in respective Gouda-derived water-soluble extracts were carried out. The identification analyses were supported with in silico calculations, i.e., heatmaps and quantitative parameters. All Gouda variants exhibited comparable ACE inhibition, whereas DPP-IV inhibition was more diversified among the samples. The samples derived from Gouda with the increased content of β-casein (both stages of ripening) had the highest DPP-IV-inhibiting potency compared to the same samples measured for ACE inhibition. Regardless of the results concerning ACE and DPP-IV inhibition among the cheese samples, the heatmap showed that the latter bioactivity was predominant in all Gouda variants, presumably because it was based on the qualitative approach (i.e., peptide presence in the sample). Our heatmap did not include the bioactivity of a single peptide as well as its quantity in the sample. In turn, the quantitative parameters showed that the best sources of ACE/DPP-IV inhibitors were all Gouda-derived extracts obtained after 60 days of the ripening. Although our protocol was efficient in showing some regularities among Gouda cheese variants, in vivo studies are recommended for more extensive investigations of this subject.     

The Causes and Potential Injurious Effects of Elevated Serum Leptin Levels in Chronic Kidney Disease Patients

Leptin is an adipokine that regulates appetite and body mass and has many other pleiotropic functions, including regulating kidney function. Increased evidence shows that chronic kidney disease (CKD) is associated with hyperleptinemia, but the reasons for this phenomenon are not fully understood. In this review, we focused on potential causes of hyperleptinemia in patients with CKD and the effects of elevated serum leptin levels on patient kidney function and cardiovascular risk. The available data indicate that the increased concentration of leptin in the blood of CKD patients may result from both decreased leptin elimination from the circulation by the kidneys (due to renal dysfunction) and increased leptin production by the adipose tissue. The overproduction of leptin by the adipose tissue could result from: (a) hyperinsulinemia; (b) chronic inflammation; and (c) significant lipid disturbances in CKD patients. Elevated leptin in CKD patients may further deteriorate kidney function and lead to increased cardiovascular risk.     

Toward a toolbox of functional block copolymers via free-radical addition of mercaptans

This work demonstrates the applicability of the free-radical addition of ω-functional mercaptans onto 1,2-polybutadienes as a modular synthetic pathway toward a toolbox of diverse functional block copolymers. Functional groups included, for instance electrolytes (carboxylic acid and amine), l-amino acid, and fluorocarbon. The number of functional groups attached to the polymer was lower than that of double bonds reacted (degree of functionalization=50–85%, typically 70–80%) due to cyclization of two neighboring units, but the narrow molecular-weight distribution of the parent (co)polymer was always maintained.     

Industry 4.0 as a Cyber-Physical System study

Advances in computation and communication are taking shape in the form of the Internet of Things, Machine-to-Machine technology, Industry 4.0, and Cyber-Physical Systems (CPS). The impact on engineering such systems is a new technical systems paradigm based on ensembles of collaborating embedded software systems. To successfully facilitate this paradigm, multiple needs can be identified along three axes: (i) online configuring an ensemble of systems, (ii) achieving a concerted function of collaborating systems, and (iii) providing the enabling infrastructure. This work focuses on the collaborative function dimension and presents a set of concrete examples of CPS challenges. The examples are illustrated based on a pick and place machine that solves a distributed version of the Towers of Hanoi puzzle. The system includes a physical environment, a wireless network, concurrent computing resources, and computational functionality such as, service arbitration, various forms of control, and processing of streaming video. The pick and place machine is of medium-size complexity. It is representative of issues occurring in industrial systems that are coming online. The entire study is provided at a computational model level, with the intent to contribute to the model-based research agenda in terms of design methods and implementation technologies necessary to make the next generation systems a reality.     

Carbon nano-onion composites: Physicochemical characteristics and biological activity

Carbon nano-onion/surfactant (CNO/surfactant) composites offer the possibility to easily produce the soluble nanostructures. That approach combines the hydrophilicity of surfactants with the robustness of carbon structures to produce composites with superior and unusual physicochemical properties. We used the following surfactants: hexadecyltrimethylammonium bromide (CTAB), sodium dodecyl sulfate (SDS), sodium dodecyl benzene sulfonate (SDBS), 4-(1,1,3,3-tetramethylbutyl)phenyl-polyethylene glycol (Triton X-100), and polyethylene glycol sorbitan monolaurate (Tween 20) to non-covalently modify CNO surfaces. The existence of stable CNO composites are clearly evidenced by direct transmission electron microscopy observations, which are also supported by thermogravimetric analyses. Dynamic light scattering and zeta potential confirmed their dispersion and stability. Additionally, the biological activity of well-dispersed CNO/surfactant composites against a strain of Escherichia coli was assayed. In vitro antimicrobial assays for the composites revealed that only the CNO/CTAB composite decreased cell viability. This activity could be assigned to the simple composite dissociation in water solutions, however antimicrobial properties of the composite are slightly better when compared with pure CTAB. This indicate some synergic effect with respect to the properties of the pure surfactant.     

Boron Clusters as Enhancers of RNase H Activity in the Smart Strategy of Gene Silencing by Antisense Oligonucleotides

Boron cluster-conjugated antisense oligonucleotides (B-ASOs) have already been developed as therapeutic agents with “two faces”, namely as potential antisense inhibitors of gene expression and as boron carriers for boron neutron capture therapy (BNCT). The previously observed high antisense activity of some B-ASOs targeting the epidermal growth factor receptor (EGFR) could not be rationally assigned to the positioning of the boron cluster unit: 1,2-dicarba-closo-dodecaborane (0), [(3,3′-Iron-1,2,1′,2′-dicarbollide) (1-), FESAN], and dodecaborate (2-) in the ASO chain and its structure or charge. For further understanding of this observation, we performed systematic studies on the efficiency of RNase H against a series of B-ASOs models. The results of kinetic analysis showed that pyrimidine-enriched B-ASO oligomers activated RNase H more efficiently than non-modified ASO. The presence of a single FESAN unit at a specific position of the B-ASO increased the kinetics of enzymatic hydrolysis of complementary RNA more than 30-fold compared with unmodified duplex ASO/RNA. Moreover, the rate of RNA hydrolysis enhanced with the increase in the negative charge of the boron cluster in the B-ASO chain. In conclusion, a “smart” strategy using ASOs conjugated with boron clusters is a milestone for the development of more efficient antisense therapeutic nucleic acids as inhibitors of gene expression.     

The Assessment of Activity of Antiseptic Agents against Biofilm of Staphylococcus aureus Measured with the Use of Processed Microscopic Images

Staphylococcal biofilms are major causative factors of non-healing wound infections. Their treatment algorithms recommend the use of locally applied antiseptic agents to counteract the spread of infection. The efficacy of antiseptics against biofilm is assessed in vitro by a set of standard quantitative and semi-quantitative methods. The development of software for image processing additionally allowed for the obtainment of quantitative data from microscopic images of biofilm dyed with propidium iodine and SYTO-9 reagents, differentiating dead cells from live ones. In this work, the method of assessment of the impact of antiseptic agents on staphylococcal biofilm in vitro, based on biofilms’ processed images, was proposed and scrutinized with regard to clinically relevant antiseptics, polyhexanide, povidone–iodine and hypochlorite. The standard quantitative culturing method was applied to validate the obtained data from processed images. The results indicated significantly higher activity of polyhexanide and povidone–iodine than hypochlorite against staphylococcal biofilm. Taking into account the fact that in vitro results of the efficacy of antiseptic agents against staphylococcal biofilm are frequently applied to back up their use in hospitals and ambulatory units, our work should be considered an important tool; providing reliable, quantitative data in this regard.     

The Detectability of the Viral RNA in Blood and Cerebrospinal Fluid of Patients with Tick-Borne Encephalitis

Background: The detection rate of viral RNA in tick-borne encephalitis (TBE) is low and variable between studies, and its diagnostic/prognostic potential is not well defined. We attempted to detect RNA of TBE virus (TBEV) in body fluids of TBE patients. Methods: We studied 98 adults and 12 children with TBEV infection, stratified by the disease phase and presentation. EDTA blood and cerebrospinal fluid (CSF) samples were obtained upon hospital admission. RNA was extracted from freshly obtained plasma, concentrated leukocyte-enriched CSF, and whole blood samples, and real time PCR was performed with a Rotor-Gene Q thermocycler. Results: TBEV RNA was detected in (1) plasma of one (of the two studied) adult patients with an abortive infection, (2) plasma of two (of the two studied) adults in the peripheral phase of TBE, and (3) plasma and blood of an adult in the neurologic phase of TBE presenting as meningoencephalomyelitis. No CSF samples were TBEV RNA-positive. Conclusions: The detection of TBEV RNA in blood might be diagnostic in the peripheral phase of TBE. The lack of TBEV RNA in the CSF cellular fraction speaks against TBEV influx into the central nervous system with infiltrating leukocytes and is consistent with a relatively low intrathecal viral burden.