The study aimed to assess the aldose reductase (AR) inhibition of selected Geranium species and determine the bioactive flavonoid constituents. Flavonoids are known to be good AR inhibitors. Among the species examined, G. sibiricum exhibited potent inhibition of AR (IC50 value, 2.4 µg/mL). Further examination of G. sibiricum, after solvent extraction and fractionation, revealed that the ethyl acetate fraction (IC50 value, 0.41 µg/mL) had a potent AR inhibitory effect. Kaempferol rhamnosides were the active compounds from this fraction. Moreover, G. sibiricum showed the highest content of kaempferol-7-O-rhamnoside and kaempferol-3,7-O-dirhamnoside among the samples examined with a concentration in the extracts of 28.1 and 2.2 mg/g, respectively. This study shows that G. sibiricum exhibits promising AR inhibitory activity, which can be explored further as a natural therapy for treating and managing complications associated with diabetes.

Milling is an essential unit operation used for particle size reduction in solid oral dosage manufacturing. The breakage of particles in a comil is due to the intense shear applied on the particles between impeller and the screen. Breakage also occurs due to the impact from a rotating impeller. Particles exit the mill based on their size relative to the aperture size of the screen bores. This study was set up to understand the working of the comil better. A new CPP (Critical Process Parameter), in the form of batch loading was identified. It was found that there are two different regimes (quasi static regime and impact regime) in which a comil generally operates, and the effect of the CPP’s (batch loading and impeller speed) on these regimes was studied. Knowledge of the effect of upstream operations on a particular unit operation is of significant importance, especially for pharmaceutical industry. For this reason, the effect of granulation variables such as liquid-to-solid ratio, gra... [more]

Oxaliplatin is a chemotherapeutic drug widely used to treat various types of tumors. However, it can induce a serious peripheral neuropathy characterized by cold and mechanical allodynia that can even disrupt the treatment schedule. Since the approval of the agent, many laboratories, including ours, have focused their research on finding a drug or method to decrease this side effect. However, to date no drug that can effectively reduce the pain without causing any adverse events has been developed, and the mechanism of the action of oxaliplatin is not clearly understood. On the dorsal root ganglia (DRG) sensory neurons, oxaliplatin is reported to modify their functions, such as the propagation of the action potential and induction of neuropathic pain. Voltage-gated sodium channels in the DRG neurons are important, as they play a major role in the excitability of the cell by initiating the action potential. Thus, in this small review, eight studies that investigated the effect of oxalip... [more]

Cell-free systems are a widely used research tool in systems and synthetic biology and a promising platform for manufacturing of proteins and chemicals. In the past, cell-free biology was primarily used to better understand fundamental biochemical processes. Notably, E. coli cell-free extracts were used in the 1960s to decipher the sequencing of the genetic code. Since then, the transcription and translation capabilities of cell-free systems have been repeatedly optimized to improve energy efficiency and product yield. Today, cell-free systems, in combination with the rise of synthetic biology, have taken on a new role as a promising technology for just-in-time manufacturing of therapeutically important biologics and high-value small molecules. They have also been implemented at an industrial scale for the production of antibodies and cytokines. In this review, we discuss the evolution of cell-free technologies, in particular advancements in extract preparation, cell-free protein synth... [more]

Cupriavidus necator DSM 545 can utilise glycerol to synthesise poly(3-hydroxybutyric acid) under unbalanced growth conditions, i.e., nitrogen limitation. To improve poly(3-hydroxybutyric acid) (PHB) batch production by C. necator through model-guided bioprocessing or genetic engineering, insights into the dynamic effect of the fermentation conditions on cell metabolism are crucial. In this work, we have used dynamic flux balance analysis (DFBA), a constrained-based stoichiometric modelling approach, to study the metabolic change associated with PHB synthesis during batch cultivation. The model employs the ‘minimisation of all fluxes’ as cellular objectives and measured extracellular fluxes as additional constraints. The mass balance constraints are further adjusted based on thermodynamic considerations. The resultant flux distribution profiles characterise the evolution of metabolic states due to adaptation to dynamic extracellular conditions and provide further insights towards improv... [more]

Following the success of and the high demand for recombinant protein-based therapeutics during the last 25 years, the pharmaceutical industry has invested significantly in the development of novel treatments based on biologics. Mammalian cells are the major production systems for these complex biopharmaceuticals, with Chinese hamster ovary (CHO) cell lines as the most important players. Over the years, various engineering strategies and modeling approaches have been used to improve microbial production platforms, such as bacteria and yeasts, as well as to create pre-optimized chassis host strains. However, the complexity of mammalian cells curtailed the optimization of these host cells by metabolic engineering. Most of the improvements of titer and productivity were achieved by media optimization and large-scale screening of producer clones. The advances made in recent years now open the door to again consider the potential application of systems biology approaches and metabolic engine... [more]

In this study, magnetic Fe3O4 nanoparticles coated with polydopamine possessing abundant amino groups (Fe3O4@PDA) were conveniently prepared, detailed, and characterized, and then firstly used as a supporting matrix for immobilizing Thermomyces lanuginosus lipase (Fe3O4@PDA@TLL). The effects of some crucial factors on the immobilization efficiency were investigated and the optimal protein loading and activity recovery were found to be 156.4 mg/g and 90.9%, respectively. Characterization studies revealed that Fe3O4@PDA@TLL displayed a broader pH and temperature adaptability as compared to the free TLL, which allows its use at wider ranges of reaction conditions. With regard to the stabilities, the immobilized TLL clearly displayed improved pH, thermal, and solvent tolerance stabilities compared to the free enzyme, suggesting that the biocompatible Fe3O4@PDA might be an outstanding material for immobilizing TLL and acting as alternative support for different enzymes.

The unique pool of features found in intracellular and extracellular bacterial biopolymers attracts a lot of research, with bacterial cellulose (BC) being one of the most versatile and common. BC is an exopolysaccharide consisting solely of cellulose, and the variation in the production process can vary its shape or even its composition when compounding is applied in situ. Together with ex situ modification pathways, including specialised polymers, particles or exclusively functional groups, BC provides a robust platform that yields complex multifunctional compounds that go far beyond ultra-high purity, intrinsic hydrophilicity, mechanical strength and biocompatibility to introduce bioactive, (pH, thermal, electro) responsive, conductive and ‘smart’ properties. This review summarises the research outcomes in BC-medical applications, focusing mainly on data from the past decade (i.e., 2010−2020), with special emphasis on BC nanocomposites as materials and devices applicable in medicine.... [more]

A class of drugs called coxibs (COX-2 inhibitors) were created to help relieve pain and inflammation of osteoarthritis and rheumatoid arthritis with the lowest amount of side effects possible. The presented paper describes a new developed, optimized and validated thin layer chromatographic (TLC)-densitometric procedure for the simultaneous assay of five coxibs: celecoxib, etoricoxib, firecoxib, rofecoxib and cimicoxib. Chromatographic separation was conducted on HPTLC F254 silica gel chromatographic plates as a stationary phase using chloroform−acetone−toluene (12:5:2, v/v/v) as a mobile phase. Densitometric detection was carried out at two wavelengths of 254 and 290 nm. The method was tested according to ICH guidelines for linearity, recovery and specificity. The presented method was linear in a wide range of concentrations for all analyzed compounds, with correlation coefficients greater than 0.99. The method is specific, precise (%RSD < 1) and accurate (more than 95%, %RSD < 2... [more]

Bioactive glass scaffolds are used in bone and tissue biomedical implants, and there is great interest in their fabrication by additive manufacturing/3D printing techniques, such as robocasting. Scaffolds need to be macroporous with voids ≥100 m to allow cell growth and vascularization, biocompatible and bioactive, with mechanical properties matching the host tissue (cancellous bone for bone implants), and able to dissolve/resorb over time. Most bioactive glasses are based on silica to form the glass network, with calcium and phosphorous content for new bone growth, and a glass modifier such as sodium, the best known being 45S5 Bioglass®. 45S5 scaffolds were first robocast in 2013 from melt-quenched glass powder. Sol−gel-synthesized bioactive glasses have potential advantages over melt-produced glasses (e.g., greater porosity and bioactivity), but until recently were never robocast as scaffolds, due to inherent problems, until 2019 when high-silica-content sol−gel bioactive glasses (H... [more]

Aquatic pollution caused by industrial effluents is an environmental issue, imposing deleterious impacts on the overall environment, specifically, on humans, by disrupting the balance of the ecosystem. Among all the industries, tanneries are considered some of the most polluting due to heavy use of toxic organic and inorganic compounds during leather processing, most of which find their way into rivers, lakes, and streams, thus exerting adverse effects on aquatic life, particularly on fish. Considering the huge concentrations of pollutants present in tannery effluents, toxicity evaluation is of prime importance. Therefore, bioassays are usually employed to assess the acute toxicity of industrial effluents and efficiency of effluent clean-up technologies as they provide a thorough response of test species to the substances present in the tested media. In the present study, the toxic effects of tannery effluent on common grass carp (Ctenopharyngodon idella) were studied for 96 h in labor... [more]

The transitioning of carbon quantum dot (cQD) applications from electrochemistry, catalysis and environmental sensing to biomedicine represents an important milestone in its 15-year history; a bellwether for its yet-unrealized potential in interventional biology, imaging, diagnostics, prophylaxis and therapy. However, despite the significant advances made over the last decade in several areas of the cQD domain, our knowledge of the exact chemical ipseity of cQDs at the Angstrom level remains either in its infancy or is largely ignored. The imminent crossing over of cQDs into biological systems and into the blood−brain barrier demands attention to the critical, yet unmet, need to resolve the inherent heterogeneity in cQD preparations and their separation into purified conformers, to identify the issues associated with potential cytotoxicity as well as to examine their bioavailability. Perhaps most importantly, and ironically neglected as well, is the compelling urgency to obtain an atom... [more]

Ensuring high quality drinking water sources is important task nowadays. To reach this task, knowledge about the impact of different chemicals on aerobic wastewater treatment is mandatory. A mixture of different chemicals reaches wastewater treatment plant every day. With the growing discharge volume of mobile toilet chemicals, active substances in these products in the past years have been recorded. The respiratory activity of activated sludge was determined to show how mobile toilet chemicals and their active substances may affect the biological wastewater treatment process. The results show negative effect of formaldehyde and bronopol on respiratory activity of activated sludge. The wastewater treatment plants influent composition and size also play important roles. Results shows that activated sludge microorganisms at a wastewater treatment plant in industrial urban area may be adapted to the higher pollutants concentration. When mobile toilet tanks are directly discharged at small... [more]

Emissions from daily and final covers of municipal solid waste (MSW) landfills can produce significant impacts on local and global environments. Simplifying, landfills can cause local impacts with odor emissions and global impacts with GHGs. This work focuses on hydrogen sulfide (H2S) and methane (CH4) emissions, with the aim of studying how it is possible to reduce their impacts by means of biofiltration systems. Both field and laboratory investigations have been carried out in Casa Rota Landfill (Tuscany, Italy). In the field trials, four pilot-scale biocovers made of compost from a source-selected organic fraction (SS compost), compost from a mechanical biological treatment plant—the residual fractions of the MSW, a mixed compost (SS-MSW compost) and sand were monitored in the daily cover area of the landfill, where high emissions were detected. Results showed that high CH4 and H2S emissions reductions occurred in the mixed SS-MSW compost plot, given a maximum methane oxidation effi... [more]

Ajmalicine is one of the most popular antihypertensive drugs obtained from the root barks of Cathranthus roseus (L.) G. Don and Rauvolfia serpentine (L.) Benth. ex Kurz. It has also potential antimicrobial, cytotoxic, central depressant and antioxidant activities. As the demand for the alkaloid is significantly high, metabolic engineering approaches are being tried to increase its production in both homologous and heterologous systems. The metabolic engineering approach requires knowledge of the metabolic regulation of the alkaloid. For understanding the metabolic regulation, fluxomic analysis is important as it helps in understanding the flux of the alkaloid through the complicated metabolic pathway. The present study was conducted to analyse the flux analysis of the ajmalicine biosynthesis, using a genetically encoded Fluorescent Resonance Energy Transfer FRET-based nanosensor for ajmalicine (FLIP-Ajn). Here, we have silenced six important genes of terpenoid indole alkaloid (TIA), na... [more]

Two-dimensional layered nanomaterial Ti3C2TX (a member of the MXene family) was used to immobilise enzyme sarcosine oxidase to fabricate a nanostructured biosensor. The device was applied for detection of sarcosine, a potential prostate cancer biomarker, in urine for the first time. The morphology and structures of MXene have been characterised by atomic force microscopy (AFM) and scanning electron microscopy (SEM). Electrochemical measurements, SEM and AFM analysis revealed that MXene interfaced with chitosan is an excellent support for enzyme immobilisation to fabricate a sensitive biosensor exhibiting a low detection limit of 18 nM and a linear range up to 7.8 µM. The proposed biosensing method also provides a short response time of 2 s and high recovery index of 102.6% for detection of sarcosine spiked into urine sample in a clinically relevant range.

Based on recent computational and experimental studies, hesperidin, a bioactive flavonoid abundant in citrus peel, stands out for its high binding affinity to the main cellular receptors of SARS-CoV-2, outperforming drugs already recommended for clinical trials. Thus, it is very promising for prophylaxis and treatment of COVID-19, along with other coexistent flavonoids such as naringin, which could help restraining the proinflammatory overreaction of the immune system. Controlled hydrodynamic cavitation processes showed the highest speed, effectiveness and efficiency in the integral and green aqueous extraction of flavonoids, essential oils and pectin from citrus peel waste. After freeze-drying, the extracted pectin showed high quality and excellent antioxidant and antibacterial activities, attributed to flavonoids and essential oils adsorbed and concentrated on its surface. This study reviews the recent evidence about hesperidin as a promising molecule, and proposes a feasible and aff... [more]

Controlled drug delivery systems can be used to carry several anticancer agents, including classical chemotherapeutic agents such as doxorubicin, paclitaxel or cisplatin, and are also used for the encapsulation of tyrosine kinase inhibitors and monoclonal antibodies. Usually, the controlled systems are used to decrease drug toxicity, increase local drug concentration or target specific organs or systems. In dogs, liposomal doxorubicin is the most known controlled drug delivery vehicle in veterinary medicine. However, several antitumor drugs can be encapsulated within these systems. Since the delivery vehicles are a relatively new topic in veterinary oncology, this review aims to discuss the current knowledge regarding the controlled drug delivery vehicles and discuss the current challenges and future direction of its use in veterinary oncology.

Microfluidics is a very facile and fast method of particulate production. Besides, it enables the manufacturing of size tuned particulate systems. Niosomes due to structural similarities have importance as alternative drug delivery systems to liposomes. Niosomes can be encapsulated or co-encapsulated with hydrophilic and lipophilic drugs. The research presented here includes the optimization of method parameters for niosome production as well as evaluation of the efficiency of microfluidics to encapsulate and co-encapsulate the drugs. For this purpose, metformin (MET) and garcinol (GC) were the model drugs. Two different non-ionic surfactants (NIS), namely Tween-20 and Span-60 with significant difference in hydrophilic-lipophilic balance (HLB) value, were chosen to analyze their efficiency to form niosomes and encapsulate one or more drugs.

Human skin has protective functions and it is a barrier that protects the interior of the body from harmful environmental factors and pathogen penetration. An important role of the skin is also to prevent the loss of water from the body and if the skin barrier is damaged, the amount of water emitted from the internal environment is increased. Therefore, it is crucial to recovery and maintenance of epidermal barrier integrity. The aim of the current work was to encapsulate Calendula officinalis flower extract in gelatin microspheres and then incorporation microspheres into thin polymeric films made from sodium alginate or mixture of sodium alginate and starch. Such materials may find applications in the cosmetic field for example in the preparation of masks for skin, according to the Calendula officinalis flower extract wide influence on skin condition. Thus, the release profile of this extract from the materials was tested under conditions corresponding to the skin (pH 5.4, 37 °C). The... [more]

Bee pollens are rich source of essential amino acids and are often considered as complete food for human beings. Herein, we exploited the potential reducing abilities of Bee pollens extract for the eco-friendly preparation of silver nanoparticles (AgNPs-G). The resulting NPs were characterized using a combination of microscopic and spectroscopic techniques. The analyses confirm the formation of spherical Ag NPs. AgNPs-G obtained from the aqueous extract of bee pollens was used to study their antibacterial properties against Gram-positive and Gram-negative microbes using the Minimum Inhibitory Concentration 50 (MIC50) method. The antibacterial properties of AgNPs-G were compared to the properties of chemically synthesized Ag NPs (AgNPs-C) using sodium borohydride as a reducing agent. The green synthesized nanoparticles (AgNPs-G) exhibited a better antibacterial activity against most of the studied strains when compared to the chemically synthesized Ag NPs (AgNPs-C). In addition, the ant... [more]

The aim of the study was to determine the use of digestate from anaerobic digestion of dairy wastewater as a culture medium for microalgae to obtain bio-oil. The experiments were conducted at a small scale in a closed raceway pond. The efficiency of the microalgae biomass production, the digestate treatment efficiency as well as the content and properties of the bio-oil obtained from the microalgal cells were analyzed. The produced biomass concentration was about 3000 ± 10.5 mg dry biomass/L, with an average growth rate of 160 ± 6.6 mgdm/L·d. The efficiency of organic compound and nutrient removal was above 90%. The bio-oil content in the biomass was about 20%. Based on the results of the study, a concept for technical-scale technology was developed.

Substrate availability plays a key role in dictating metabolic strategies. Most microorganisms consume carbon/energy sources in a sequential, preferential order. The presented study investigates metabolic strategies of Alicyclobacillus acidocaldarius, a thermoacidophilic bacterium that has been shown to co-utilize glucose and xylose, as well as degrade phenolic compounds. An existing metabolic model was expanded to include phenol degradation and was analyzed with both metabolic pathway and constraint-based analysis methods. Elementary flux mode analysis was used in conjunction with resource allocation theory to investigate ecologically optimal metabolic pathways for different carbon substrate combinations. Additionally, a dynamic version of flux balance analysis was used to generate time-resolved simulations of growth on phenol and xylose. Results showed that availability of xylose along with glucose did not predict enhanced growth efficiency beyond that of glucose alone, but did predi... [more]

The aim of this study was preparation of new derivatives based on 2-((4-chlorophenoxy)methyl)-N-(arylcarbamothioyl)benzamide structure; the new compounds were characterized by IR, NMR (1H, 13C) spectroscopy, and elemental analysis. The obtained compounds were evaluated for their in vitro antimicrobial activity against planktonic and biofilm-embedded microbial cells (Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa, Candida albicans), by qualitative and quantitative assays. Some of the compounds revealed promising antibacterial and antifungal activities, with low minimum inhibitory concentration values between 0.15 and 2.5 mg/mL and minimal biofilm eradication concentrations of 0.019−2.5 mg/mL. To investigate the potential target of their antibacterial activity, in silico drug-likeness and molecular docking screenings on Staphylococcus aureus DNA gyrase were performed. The compound with the best antibacterial activity (1g) was docked into topoisomer... [more]

Tissue adhesives have been widely used in surgical procedures. Compared to traditional surgical sutures, tissue adhesives provide fast bonding experiences and full closure of wounds. However, current tissue adhesives are mostly fossil-based synthetic products. Therefore, it is of great significance to explore the use of natural materials in tissue adhesives. Whey is a low-end byproduct of cheese manufacturing. Whey protein, a group of small globular proteins, can exhibit adhesive properties if their structures are modified by physical or chemical means. The objectives of this study were to investigate the functional and structural properties of whey protein-based tissue adhesive, along with the antibacterial effect of totarol, a natural antimicrobial agent. Whey protein isolate (WPI) solutions (25%−33% protein) were mixed with different levels (0.1%−0.3% w/w) of totarol. The mixtures were analyzed for total plate count and yeast and mold count. The lap-shear bonding strength was tested... [more]