4-Hydroxybenzoic Acid-Based Hydrazide-Hydrazones as Potent Growth Inhibition Agents of Laccase-Producing Phytopathogenic Fungi That Are Useful in the Protection of Oilseed Crops.

The research on new compounds against plant pathogens is still socially and economically important. It results from the increasing resistance of pests to plant protection products and the need to maintain high yields of crops, particularly oilseed crops used to manufacture edible and industrial oils and biofuels. We tested thirty-five semi-synthetic hydrazide-hydrazones with aromatic fragments of natural origin against phytopathogenic laccase-producing fungi such as Botrytis cinerea, Sclerotinia sclerotiorum, and Cerrena unicolor. Among the investigated molecules previously identified as potent laccase inhibitors were also strong antifungal agents against the fungal species tested. The highest antifungal activity showed derivatives of 4-hydroxybenzoic acid and salicylic aldehydes with 3-tert-butyl, phenyl, or isopropyl substituents. S. sclerotiorum appeared to be the most susceptible to the tested compounds, with the lowest IC50 values between 0.5 and 1.8 µg/mL. We applied two variants of phytotoxicity tests for representative crop seeds and selected hydrazide-hydrazones. Most tested molecules show no or low phytotoxic effect for flax and sunflower seeds. Moreover, a positive impact on seed germination infected with fungi was observed. With the potential for application, the cytotoxicity of the hydrazide-hydrazones of choice toward MCF-10A and BALB/3T3 cell lines was lower than that of the azoxystrobin fungicide tested.

A catalytic membrane approach as a way to obtain sweet and unsweet lactose-free milk.

The growing need in the current market for innovative solutions to obtain lactose-free (L-F) milk is caused by the annual increase in the prevalence of lactose intolerance inside as well as the newborn, children, and adults. Various configurations of enzymes can yield two distinct L-F products: sweet (ß-galactosidase) and unsweet (ß-galactosidase and glucose oxidase) L-F milk. In addition, the reduction of sweetness through glucose decomposition should be performed in a one-pot mode with catalase to eliminate product inhibition caused by H2O2. Both L-F products enjoy popularity among a rapidly expanding group of consumers. Although enzyme immobilization techniques are well known in industrial processes, new carriers and economic strategies are still being searched. Polymeric carriers, due to the variety of functional groups and non-toxicity, are attractive propositions for individual and co-immobilization of food enzymes. In the presented work, two strategies (with free and immobilized enzymes; ß-galactosidase NOLA, glucose oxidase from Aspergillus niger, and catalase from Serratia sp.) for obtaining sweet and unsweet L-F milk under low-temperature conditions were proposed. For free enzymes, achieving the critical assumption, lactose hydrolysis and glucose decomposition occurred after 1 and 4.3 h, respectively. The tested catalytic membranes were created on regenerated cellulose and polyamide. In both cases, the time required for lactose and glucose bioconversion was extended compared to free enzymes. However, these preparations could be reused for up to five (ß-galactosidase) and ten cycles (glucose oxidase with catalase).

Hyaluronic Acid-Alginate Homogeneous Structures with Polylactide Coating Applied in Controlled Antibiotic Release.

The use of a controlled-release drug carrier is an innovative solution for the treatment of local infections, in particular in dentistry, skin diseases, and in open wounds. The biocompatibility, biodegradability, the possibility of a large amount of drug adsorbed (especially those with hydrophilic properties), and the ability to create structures of any shape and size are the reasons for hydrogels to be frequently studied. The main disadvantage of hydrogel carriers is the rapid rate of drug release; hence, in this study, an attempt was made to additionally chemically cross-link 1-ethyl-3-(3-dimethyl aminopropyl)-1-carbodiimide hydrochloride (EDC) with the hyaluronic acid-alginate (HA-SAL) structure. The answer to significantly reduce the mass flux typical for hydrogel structure was to surround it with a polymer layer using a dry cover. By coating the carriers with polylactide, the release time was increased by around forty times. As the carriers were designed to reduce local bacterial infections, among others in dentistry, the released antibiotics were amoxycillin, metronidazole, and doxycycline.

Critical Parameters in an Enzymatic Way to Obtain the Unsweet Lactose-Free Milk Using Catalase and Glucose Oxidase Co-Encapsulated into Hydrogel with Chemical Cross-Linking.

The presented work involves obtaining and characterising a two-enzymatic one-pot bioreactor, including encapsulated (co-immobilised) glucose oxidase and catalase. The enzymatic capsules were applied to produce unsweet, lactose-free milk during low-temperature catalysis. Furthermore, operational conditions, like pH and aeration, were selected in the paper, which sorts out discrepancies in literature reports. All experiments were carried out at 12 °C, corresponding to milk storage and transportation temperature. Preliminary studies (for reasons of analytical accuracy) were carried out in a buffer (pH, concentration of sugars mimicking conditions in the lactose-free milk, the initial glucose concentration 27.5 g/L) verified by processes carried out in milk in the final stage of the study. The presented results showed the need for regulating pH and the aeration of the reaction mixture in the continuous mode during the process. The procedure of co-immobilisation was performed in an alginate matrix with the cross-linking of glutaraldehyde or carbodiimide while carbodiimide showed better enzymes retention inside alginate capsules. Co-encapsulated enzymes could be used for nine cycles, preserving finally about 40% of the initial activity.

Low- and High-Pressure Membrane Separation in the Production of Process Water for Coke Quenching.

Although the time for operating mines and coking plants in many countries is coming to an end due to climate change, we must still ensure that the pollution generated by this source of the economy is minimized. Despite the several stages of treatment of the coke-oven effluent, completed with nitrification and denitrification processes preceding final sedimentation, the stream obtained does not meet the requirements of water for coke quenching. That is why the stream after biodegradation and sedimentation was treated on membrane units to ensure water reusing in the coking plant. As the subjected stream contained both solid and dissolved pollutants, a two-stage system was proposed: low- and high-pressure membrane filtration. Industrial modules were tested on pilot units operating under industrial plant conditions. In the case of the ultrafiltration process, all the tested ultrafiltration modules fulfilled the primary task. All of them separated almost completely the turbidities present in the stream, which would have disturbed the operation of the high-pressure plant. Considering the decrease in permeate flux and the possibility of cleaning, a PCI membrane made of PVDF tubes with a diameter of 12.5 mm and pore size of 20 µm was selected. Regarding the high-pressure membrane filtration, the reverse osmosis membrane was significantly better in the removal efficiency of both organic and inorganic dissolved substances. An operating pressure of 3 MPa was chosen for the system. Hence, membrane processes, which are not used as stand-alone treatment units for coke-oven effluents, function well as a final treatment stage.

Polymeric Microfiltration Membranes Modification by Supercritical Solvent Impregnation-Potential Application in Open Surgical Wound Ventilation.

This study investigated supercritical solvent impregnation of polyamide microfiltration membranes with carvacrol and the potential application of the modified membranes in ventilation of open surgical wounds. The impregnation process was conducted in batch mode at a temperature of 40 °C under pressures of 10, 15, and 20 MPa for contact times from 1 to 6 h. FTIR was applied to confirm the presence of carvacrol on the membrane surface. In the next step, the impact of the modification on the membrane structure was studied using scanning electron and ion beam microscopy and cross-filtration tests. Further, the release of carvacrol in carbon dioxide was determined, and finally, an open thoracic cavity model was applied to evaluate the efficiency of carvacrol-loaded membranes in contamination prevention. Carvacrol loadings of up to 43 wt.% were obtained under the selected operating conditions. The swelling effect was detectable. However, its impact on membrane functionality was minor. An average of 18.3 µg of carvacrol was released from membranes per liter of carbon dioxide for the flow of interest. Membranes with 30-34 wt.% carvacrol were efficient in the open thoracic cavity model applied, reducing the contamination levels by 27% compared to insufflation with standard membranes.

Drug Carriers Based on Graphene Oxide and Hydrogel: Opportunities and Challenges in Infection Control Tested by Amoxicillin Release.

Graphene oxide (GO) was proposed as an efficient carrier of antibiotics. The model drug, amoxicillin (AMOX), was attached to GO using a peptide linker (Leu-Leu-Gly). GO-AMOX was dispersed in a hydrogel to which the enzyme responsible for releasing AMOX from GO was also added. The drug molecules were released by enzymatic hydrolysis of the peptide bond in the linker. As the selected enzyme, bromelain, a plant enzyme, was used. The antibacterial nature of the carrier was determined by its ability to inhibit the growth of the Enterococcus faecalis strain, which is one of the bacterial species responsible for periodontal and root canal diseases. The prepared carrier contained only biocompatible substances, and the confirmation of its lack of cytotoxicity was verified based on the mouse fibrosarcoma cell line WEHI 164. The proposed type of preparation, as a universal carrier of many different antibiotic molecules, can be considered as a suitable solution in the treatment of inflammation in dentistry.

Dynamic modeling of the activated sludge microbial growth and activity under exposure to heavy metals.

The presence of heavy metals in the environment can lead to ecological and health problems. The evolution of biological systems, such as activated sludge, exposed to heavy metals is still underexplored. Therefore, this study sought to develop a model of microorganism activity and growth in activated sludge and used it to investigate the toxicity of five metals: Cu, Cd, Ni, Zn, and Ag. Patterns in the evolution of the toxic effects caused by these metals were similar at the beginning of exposure. Differences in toxicity between metal ions were noted for longer exposure times. Changes in model parameters indicate the influence of metal ions on the mass and energy balance of living cells. Decreases in new enzyme units and biomass production yields in contaminated activated sludge indicate a shift from anabolic reactions to metal homeostasis and resistance.

Drinking Behavior, Taste Preferences and Special Beer Perception among Romanian University Students: A Qualitative Assessment Research.

The transition from adolescence to adulthood can be a challenging period for many students. This period is associated with an increase in alcohol consumption (AC) which can develop a drinking behavior or shape the preferences for certain alcoholic beverages. The purpose of this study was to analyze the AC pattern among Romanian university students, by investigating the association between taste and consumption, including preferences for special beer. A 30-item omnibus-type questionnaire was distributed to undergraduate students and used to gather sociodemographic data, alcohol expectancies, drinking motives and consequences, and special beer consumption. Results showed a statistically significant relationship between the age of first alcohol use and the existence of an alcoholic family member. The main reasons for AC are taste, sensation, relaxation, and socialization. Both female and male students tend to drink occasionally, with a preference for public places. Female students prefer a sweet taste, choosing special beers over the regular ones. The students' residence may also influence the choice of special beers. Understanding the students' drinking behavior and taste preferences is essential to create useful strategies to discourage excessive AC. Special beer, a growing segment in the beverage industry, could represent a healthier and safety alternative to AC.

Functional Modification of Cellulose Acetate Microfiltration Membranes by Supercritical Solvent Impregnation.

This study investigates the modification of commercial cellulose acetate microfiltration membranes by supercritical solvent impregnation with thymol to provide them with antibacterial properties. The impregnation process was conducted in a batch mode, and the effect of pressure and processing time on thymol loading was followed. The impact of the modification on the membrane's microstructure was analyzed using scanning electron and ion-beam microscopy, and membranes' functionality was tested in a cross-flow filtration system. The antibiofilm properties of the obtained materials were studied against Staphyloccocus aureus and Pseudomonas aeruginosa, while membranes' blocking in contact with bacteria was examined for S. aureus and Escherichia coli. The results revealed a fast impregnation process with high thymol loadings achievable after just 0.5 h at 15 MPa and 20 MPa. The presence of 20% of thymol provided strong antibiofilm properties against the tested strains without affecting the membrane's functionality. The study showed that these strong antibacterial properties could be implemented to the commercial membranes' defined polymeric structure in a short and environmentally friendly process.

Cellulose Acetate Membranes Modification by Aminosilane Grafting in Supercritical Carbon Dioxide towards Antibiofilm Properties.

The study explores the grafting of cellulose acetate microfiltration membranes with an aminosilane to attain antibiofilm properties. The grafting reaction was performed in the supercritical carbon dioxide used as a transport and reaction medium. The FTIR analyses and dissolution tests confirmed the covalent bonding between the aminosilane and polymer. The membranes' microstructure was investigated using a dual-beam SEM and ion microscopy, and no adverse effects of the processing were found. The modified membranes showed a more hydrophilic nature and larger water permeate flow rate than the neat cellulose acetate membranes. The tests in a cross-filtration unit showed that modified membranes were considerably less blocked after a week of exposure to Staphylococcus aureus and Escherichia coli than the original ones. Microbiological investigations revealed strong antibiofilm properties of the grafted membranes in experiments with Staphylococcus aureus, Listeria monocytogenes, Escherichia coli, and Salmonella Enteritidis.

Three-Stage Membrane Treatment of Wastewater from Biodiesel Production-Preliminary Research.

As biodiesel production as renewable fuel increases, so does the amount of wastewater resulting from this technology. Wastewater is generated during the so-called biodiesel washing, i.e., washing out glycerol and methanol with water. The purified biodiesel must meet international standards, such as EN 14214 or the American ASTM D6751 standard. To fully say that biodiesel technology is environmentally friendly, the amount of wastewater should be minimized. It is also desirable that the purified water can be recycled to the technology. For this purpose, wastewater pre-treated by flotation, during which mainly oils are removed, was subjected to three-stage membrane separation. For each of the stages, the membrane was selected and characterized in terms of its separation capacity and stream stability. Starting with microfiltration, which was mainly aimed at reducing turbidity, affects the permeate flux in the following steps. Then, ultrafiltration and nanofiltration membranes were selected. These membranes were aimed at reducing the concentration of inorganic and organic substances. Consequently the cascade was composed of: MF-0.45 µm, UF-150 kDa, and NF-characterized by an 80% degree of desalination. The final permeate has a salt concentration of less than 0.15 g/L and can be reused in biodiesel technology.

Brewer's Spent Grains-Valuable Beer Industry By-Product.

The brewing sector is a significant part of the global food industry. Breweries produce large quantities of wastes, including wastewater and brewer's spent grains. Currently, upcycling of food industry by-products is one of the principles of the circular economy. The aim of this review is to present possible ways to utilize common solid by-product from the brewing sector. Brewer's spent grains (BSG) is a good material for sorption and processing into activated carbon. Another way to utilize spent grains is to use them as a fuel in raw form, after hydrothermal carbonization or as a feedstock for anaerobic digestion. The mentioned by-products may also be utilized in animal and human nutrition. Moreover, BSG is a waste rich in various substances that may be extracted for further utilization. It is likely that, in upcoming years, brewer's spent grains will not be considered as a by-product, but as a desirable raw material for various branches of industry.

Graphene oxide as a potential drug carrier - Chemical carrier activation, drug attachment and its enzymatic controlled release.

Graphene oxide (GO), due to its properties, such as nanometric dimensions, large specific surface area, and biocompatibility, can be used as a carrier in controlled drug release systems. The method of its chemical activation before drug molecules binding was elaborated. Doxorubicin (DOX), an anticancer drug, was attached to the surface of GO via the Gly-Gly-Leu linker. Approximately 3.07 · 1020 molecules of the tripeptide were attached to 1 g of GO and subsequently almost the same number of DOX molecules. GO was suspended inside a sol surrounded by a thin porous membrane. The bound DOX was effectively released using thermolysin, an enzyme cleaving peptide bonds between Gly and Leu inside the linker structure. The membrane, as the shell was responsible for keeping enzyme molecules in their native form and GO flakes inside the carrier, simultaneously allowed the released drug molecules to diffuse outside. The rate of drug release was described as a function of the enzyme concentration and mass of DOX expressed on carrier volume; thus, the daily dose and length of the therapy can be controlled. Studies involving the cell line of mice fibrosarcoma WEHI 164 have shown that the prepared carrier itself is not toxic and only the introduction of DOX-releasing enzyme into it causes cell death.

Synthesis and Structure-Activity Relationship Studies of Hydrazide-Hydrazones as Inhibitors of Laccase from Trametes versicolor.

A series of hydrazide-hydrazones 1-3, the imine derivatives of hydrazides and aldehydes bearing benzene rings, were screened as inhibitors of laccase from Trametes versicolor. Laccase is a copper-containing enzyme which inhibition might prevent or reduce the activity of the plant pathogens that produce it in various biochemical processes. The kinetic and molecular modeling studies were performed and for selected compounds, the docking results were discussed. Seven 4-hydroxybenzhydrazide (4-HBAH) derivatives exhibited micromolar activity Ki = 24-674 µM with the predicted and desirable competitive type of inhibition. The structure-activity relationship (SAR) analysis revealed that a slim salicylic aldehyde framework had a pivotal role in stabilization of the molecules near the substrate docking site. Furthermore, the presence of phenyl and bulky tert-butyl substituents in position 3 in salicylic aldehyde fragment favored strong interaction with the substrate-binding pocket in laccase. Both 3- and 4-HBAH derivatives containing larger 3-tert-butyl-5-methyl- or 3,5-di-tert-butyl-2-hydroxy-benzylidene unit, did not bind to the active site of laccase and, interestingly, acted as non-competitive (Ki = 32.0 µM) or uncompetitive (Ki = 17.9 µM) inhibitors, respectively. From the easily available laccase inhibitors only sodium azide, harmful to environment and non-specific, was over 6 times more active than the above compounds.

Enzymatic Hydrolysis of Hyaluronic Layer before Drug Dosing.

BACKGROUND: The local therapy is a particularly proper approach in treatment of cancer diseases, at which the drugs are highly toxic. Unfortunately, antitumor drugs diffusion is quite often limited by hyaluronic acid layers abundantly found in a cancer tissue. METHODS: The undertaken approach assumed relaxation of hyaluronan layer by using a hydrolytic enzyme - hyaluronidase. The considered process corresponds to the heterogenic catalysis with substrate occurred as a layer and a catalyst in a native form. It was shown that enzyme concentration around 4 mg/L kept for 10 minutes in body fluid surrounding HA layers is enough to allow the drug to diffuse. RESULTS: Cyanocobalamin, as a model drug, was applied in diffusion processes testing. The possibility of hyaluronidase releasing in the place close to hyaluronic acid layers was also considered. Hydrogel carriers promoting fast release of hyaluronidase were characterized. The profile of releasing Hase from the tested carriers based on κ-carrageenan or sodium alginate with polyvinyl alcohol corresponded to Higuchi model. CONCLUSION: An expected rate value (the mass transport coefficient app. 4.0.10-7 m/s) was obtained for blends 0.5% κ-carrageenan, 4% polyvinyl alcohol and 0.5% sodium alginate, 6% polyvinyl alcohol.