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Biol. Life Sci. Forum, 2022, IECBM 2022

Number of Papers: 34
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Cover Story (view full-size image): This volume brings together the complete presentations of the 2nd International Electronic Conference on Biomolecules: Biomacromolecules and the Modern World Challenge (IECBM 2022) held on 1–15 [...] Read more.
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Abstract
The First Opsins Prediction in Achatina fulica Genome
Biol. Life Sci. Forum 2022, 20(1), 1; https://doi.org/10.3390/IECBM2022-13518 - 11 Nov 2022
Viewed by 464
Abstract
The variety of spectral properties of visual pigments is provided mainly by the amino acid sequences of their protein component, the so-called visual opsins. However, such data on these proteins of gastropod mollusks are very scarce, and for terrestrial species are absent altogether. [...] Read more.
The variety of spectral properties of visual pigments is provided mainly by the amino acid sequences of their protein component, the so-called visual opsins. However, such data on these proteins of gastropod mollusks are very scarce, and for terrestrial species are absent altogether. Meanwhile, the first sequenced genome of the terrestrial gastropod, Achatina fulica, has been published. So, the aim of this study is the prediction of the amino acid sequences of opsins in the A. fulica genome. To this purpose, all known mollusk opsin sequences from the NCBI (National Center for Biotechnology Information) Protein database were collected. To analyze them, we performed a multiple sequence alignment in Unipro UGENE software (version 39.0) (Unipro, Russia) by Clustal Omega and refined it by MUSCLE algorithm, both integrated in UGENE, and then constructed the HMM (Hidden Markov Models) profile, which was used to predict opsins in the A. fulica genome. The prediction of opsin sequences was performed using HMMER software (version 3.3.2). To confirm that the obtained sequences belong to opsins, we searched for the following conserved domain: 7tmA_Opsin_Gq_invertebrates. As a result, we determined five rhodopsin G-coupled protein sequences in the A. fulica genome: Afu011750, Afu004575, Afu005002, Afu003765, and Afu022267. This is the first attempt to annotate the Achatina fulica genome, which was initiated by searching for opsins. Our study opens up new opportunities for further, more detailed evolutionary and genetic studies of mollusks, including the study of their visual mechanisms. However, it should be noted that opsins form a diverse multifunctional group of proteins, of which only some comprise visual pigments. Therefore, work towards the identification of visual opsins in Achatina fulica should be continued. Full article
Abstract
Biotechnological Complex Based on Lipase and β-Cyclodextrin on Hydrolysis of Acylglycerides in Plant Oils and Fats
Biol. Life Sci. Forum 2022, 20(1), 2; https://doi.org/10.3390/IECBM2022-13404 - 01 Nov 2022
Viewed by 368
Abstract
The present study aimed to evaluate in vitro enzymatic activity of a novel biotechnological active complex based on natural origin compounds—thermophilic lipase and β-cyclodextrin—for hydrolysis of acylglycerides in plant oils and fats. β-cyclodextrin (β-CD) as an additive has attracted attention for its enhanced [...] Read more.
The present study aimed to evaluate in vitro enzymatic activity of a novel biotechnological active complex based on natural origin compounds—thermophilic lipase and β-cyclodextrin—for hydrolysis of acylglycerides in plant oils and fats. β-cyclodextrin (β-CD) as an additive has attracted attention for its enhanced stability and efficiency of enzymes. In present study, the effects of β-CD on enzymatic hydrolysis of acylglycerides by thermophilic lipase were investigated by modern methods. The UV-spectroscopy, electron microscopy with TEM and kinetics of enzymatic hydrolysis were compared by the addition of β-CD. The results showed that lipase could produce the highest yield of oleic acid in presence of β-CD after 1 and 3 h. The UV spectroscopy demonstrated that the absorbance and fluorescence of lipase decreased with increasing concentration of β-CD due to surface interaction and change of enzyme configuration. Moreover, electron microscopy with TEM showed that lipase formed a special active conglomerate with β-CD for improving hydrolysis and stability. Dermatology evaluation indicated that this complex, when added to household products, did not affect sensitive skin of hands. Overall results indicate that β-CD could increase enzymatic activity of the lipase against acylglycerides and can be considered as promising composition in ecological household products for regular hand application. Full article
Abstract
Film-Based Hydrogel Designed as Functional Biomaterial for Treatment of Skin Wound
Biol. Life Sci. Forum 2022, 20(1), 3; https://doi.org/10.3390/IECBM2022-13508 - 09 Nov 2022
Viewed by 406
Abstract
Multidrug-resistant bacteria can lead to skin wound infections, pain and long-term treatment. The development of high-performance dressings for the treatment of infected skin wounds represents a necessary requirement. In this paper it was designed and synthesized adhesive hydrogel films containing active agents such [...] Read more.
Multidrug-resistant bacteria can lead to skin wound infections, pain and long-term treatment. The development of high-performance dressings for the treatment of infected skin wounds represents a necessary requirement. In this paper it was designed and synthesized adhesive hydrogel films containing active agents such as hyaluronic acid (HA) and lidocaine, as local anesthetic. HA is an extremely effective and long-lasting moisturizer, biocompatible, with a regenerating effect, promoting the regeneration of damaged skin. lidocaine has been incorporated to relieve acute pain and improve patient comfort and endurance. Films without HA and lidocaine were also produced as control sample. The successful synthesis of films obtained at room temperature, from aqueous solutions, was also confirmed by Fourier transform infrared analysis, X-ray diffraction and scanning electron analysis. Water absorption, adhesion and mechanical strength of the films-based hydrogel are improved with the introduction of HA and lidocaine, leading to the rapid skin wound healing process. The obtained materials will be used as promising materials able to restore the structural and functional properties of the skin. Wet adhesive backing films with good adhesion to skin, encapsulating HA with different molecular weights and lidocaine were developed to explore their potential to be used as a patch for painless treatment and healing of skin wounds. The cytocompatibility studies confirmed that the obtained films-based hydrogel have demonstrated no cytotoxic effect. Full article
Abstract
Co-Expression of VEGF and FGF2 Mediated by Multigenic Plasmid Constructs Promotes Blood Flow Restoration in a Rat Model of Hind Limb Ischemia
Biol. Life Sci. Forum 2022, 20(1), 4; https://doi.org/10.3390/IECBM2022-13517 - 11 Nov 2022
Viewed by 459
Abstract
Peripheral arterial disease (PAD) is characterized by stenosis and occlusion of the arteries leading to poor blood supply to the limb. Patients with PAD suffer pain at rest, intermittent claudication, skin ulcers, and gangrene. The end-stage of the disease could lead to limb [...] Read more.
Peripheral arterial disease (PAD) is characterized by stenosis and occlusion of the arteries leading to poor blood supply to the limb. Patients with PAD suffer pain at rest, intermittent claudication, skin ulcers, and gangrene. The end-stage of the disease could lead to limb amputation despite optimal medical and surgical management. The delivery of angiogenic factors to restore tissue perfusion is an attractive strategy, both as a primary and adjunctive treatment for PAD. We synthesized multigenic plasmid constructs expressing combinations of VEGF, FGF2, and DsRed genes:pVax1-VEGF-FGF2-DsRed, pVax1-VEGF-DsRed, pVax1-FGF2-DsRed, and pVax1-DsRed. In the constructed vectors, gene sequences are linked through the furin-containing 2A-peptide sequence of picornaviruses. Plasmid vector pVax1 is approved by the FDA for use in clinical trials. Previously, we confirmed the functionality of the developed non-viral constructs and the synthesis of VEGF, FGF2, and DsRed proteins by transfected cells. At this stage, we injected plasmid constructs into rat muscles after hind limb ischemia. Quantitative analysis of serum cytokines and chemokines of experimental and control groups on 3, 14, and 21 days after plasmids injection showed no significant differences in the secretion of the 18 cytokines studied. We observed a gradual increase in volumetric blood flow in the experimental groups, despite decreased expression of VEGF and FGF2 on 14 and 21 days. On day 21, the maximum increase in volumetric blood flow was in the pVax1-VEGF-FGF2-DsRed group. In turn, the maximum number of capillaries at 21 days was in the pVax1-VEGF-DsRed group. Capillary density was increased in pVax1-VEGF-FGF2-DsRed and pVax1-FGF2-DsRed groups compared to control groups. We also observed low expression levels of caspase-3 and caspase-9 in the muscles of the experimental groups. Thus, co-expression of VEGF and FGF2 stimulates angiogenic and regenerative processes in a rat model of hind limb ischemia. In addition, the results of this study are consistent with our previous work and confirm the effectiveness of using systems based on 2A-peptide sequences for transgene co-expression. The study of the serum cytokine profile showed the absence of adverse immune effects, indicating the safety of the non-viral constructs used. These results suggest the possibility of using these non-viral constructs to enhance therapeutic angiogenesis in the treatment of ischemic diseases. This paper has been supported by the Kazan Federal University Strategic Academic Leadership Program (PRIORITY-2030). Full article
Abstract
Structure, Properties and Biological Activity of Chitosan Salts with L- and D-Aspartic Acid
Biol. Life Sci. Forum 2022, 20(1), 5; https://doi.org/10.3390/IECBM2022-13399 - 01 Nov 2022
Viewed by 370
Abstract
A comprehensive study of the structure, properties and biological functionality of salt chitosan complexes with L- and D-aspartic acid (AspA) was carried out. It has been established that these polymer salts differ in their spatial organization, chirooptic characteristics, surface charge and macrocoil size. [...] Read more.
A comprehensive study of the structure, properties and biological functionality of salt chitosan complexes with L- and D-aspartic acid (AspA) was carried out. It has been established that these polymer salts differ in their spatial organization, chirooptic characteristics, surface charge and macrocoil size. In experiments in vitro on a wide range of biological objects (unicellular algae, planktonic crustaceans, aerobic bacterial microorganisms, cell cultures and test plants), it was found that the chitosan salt with D-AspA exhibited the best biological activity. The results obtained confirm our hypothesis that the biological homochiral hierarchy principles are most consistent with the chitosan (D-aminoglycan) derivatives with the D-antipode of the acid. Full article
Abstract
Effect of Ohmic Heating Nixtamalization on the Structural and Physicochemical Characteristics of Instant Maize Flours and Their Relation to Starch Modifications
Biol. Life Sci. Forum 2022, 20(1), 7; https://doi.org/10.3390/IECBM2022-13380 - 01 Nov 2022
Cited by 1 | Viewed by 345
Abstract
This study investigated the changes in the physicochemical properties of maize starch on nixtamalized flours produced with ohmic heating (OH). Samples were prepared using the following OH process variables and levels: cooking temperature (85 and 90 °C), heating time (0, 5 and 10 [...] Read more.
This study investigated the changes in the physicochemical properties of maize starch on nixtamalized flours produced with ohmic heating (OH). Samples were prepared using the following OH process variables and levels: cooking temperature (85 and 90 °C), heating time (0, 5 and 10 min) and voltage (120 and 130 V). Changes were studied using their viscosity profiles, differential scanning calorimetry (DSC), X-ray diffraction (XRD) and scanning electron microscopy (SEM). Results indicated that flour viscosity was affected by increasing time and/or temperature, but also by greater electrical fields. This was due to gelatinization and electroporation, shown as damage of the starch granule in SEM. DSC and XRD indicated not only gelatinization and loss of crystalline structures, but also formation of new amylose–lipid interactions, stabilizing the starch system and causing lower peak viscosity. Full article
Abstract
A Beneficial Bio-Waste with a Zero-Waste Approach: Peanut Shell
Biol. Life Sci. Forum 2022, 20(1), 8; https://doi.org/10.3390/IECBM2022-13381 - 01 Nov 2022
Viewed by 389
Abstract
The recycling of food and agricultural waste, which is released as a result of domestic and agricultural uses, instead of throwing them into the garbage cycle and the environment, is of great importance both for the protection of the environment and the minimization [...] Read more.
The recycling of food and agricultural waste, which is released as a result of domestic and agricultural uses, instead of throwing them into the garbage cycle and the environment, is of great importance both for the protection of the environment and the minimization of other environmental pollutants. In recent years, human population growth, pandemic developments (COVID-19), climate change and global warming have increased significantly. These increases endanger environmental health. Therefore, researchers are investigating different alternatives in terms of both human and environmental health. This paper evaluates the possible use of the shell part of the peanut, which is a food with high nutritional value. Peanut (Arachys hypogaea) is a plant from the Fabaceae family. Peanut is a valuable food product with a wide range of uses all over the world. Their shells are an indispensable part of the garbage cycle and have a fibrous and lignocellulosic (cellulose content: 45%, hemicellulose content: 6%, lignin content: 36%) structure. In addition, it has a very slow degradation rate under natural conditions, which is a great advantage for other wastes. Today, most peanut shells are disposed of by incineration and burial, which causes environmental pollution. For this reason, this waste should be used in various sectors with a zero waste approach. Increasing environmental pollution all over the world day by day, unconscious energy consumption and climate change have led countries to seek alternative solutions for environmental issues and to develop environmentally friendly-technological methods. Peanut shell is used intensively in fields such as compost material, energy sector (biofuel, biodiesel, CO2 emission reduction, etc.), cosmetics (nail polish, lipstick, etc.), soil improvement, drinking water and wastewater treatment (adsorbent, nanomaterial, filter etc.). In this study, the use of peanut shells from Osmaniye province as an environmentally friendly, economical and easily available biosorbent was investigated. Full article
Abstract
Design of Enzyme Stabilization Systems for Gas Separation: Novel Studies on Formation of Enzyme Based W/O Emulsions by Direct Membrane Emulsification to Synthesise Emulsion-Based Supported Liquid Membrane for CO2 Capture
Biol. Life Sci. Forum 2022, 20(1), 9; https://doi.org/10.3390/IECBM2022-13389 - 01 Nov 2022
Viewed by 358
Abstract
Membrane-based gas separation is an important unit operation in chemical industries due to its simplicity, ease of operation, reduced energy consumption, and compact structure. For gas separation, novel studies were carried out by synthesising enzyme-stabilised systems consisting of emulsion-based supported liquid membranes (E-SLMs) [...] Read more.
Membrane-based gas separation is an important unit operation in chemical industries due to its simplicity, ease of operation, reduced energy consumption, and compact structure. For gas separation, novel studies were carried out by synthesising enzyme-stabilised systems consisting of emulsion-based supported liquid membranes (E-SLMs) the pores of which pores were impregnated with water-in-oil (W/O) emulsions produced by direct membrane emulsification. This technique has gained attention, as it consumes low energy and is mild and suitable for sensitive enzymes. This case study involves the capture of CO2 by the enzyme carbonic anhydrase (CA). The composition of the oil phase was optimised amongst various edible oils, aiming for the one with the highest CO2 sorption capability. The water phase was optimised based on the stability of the CA enzyme in the aqueous phase in the presence of various surfactants and their concentrations. The optimised emulsions consisted of 2% Tween 80 (w/w) in corn oil as the continuous phase and 0.5 g L−1 CA enzyme with 5% PEG300 (w/w) in aqueous solution as the dispersed phase. The emulsions were prepared with a Microdyn Nadir UP150 polymeric membrane. These emulsions were impregnated onto a hydrophobic PVDF membrane to prepare E-SLM. For comparative studies, liquid membranes were also prepared without the CA enzyme in the emulsions, and a supported liquid membrane (SLM) was prepared by impregnating corn oil onto the membrane. Lastly, the permeabilities of the main components of biogas, CO2, and CH4, through the SLM and E-SLMs, were evaluated. The permeability of CO2 increased (~15%) and CH4 decreased (~60%) through the E-SLM containing CA when compared to the SLM and E-SLM without CA. Subsequently, the selectivity of CO2 increased in the presence of low concentration of CA. This work suggests the enhanced, synergetic effects of carbonic anhydrase within a bio-based emulsion system for CO2 capture. Full article
Abstract
A Molecular Dynamics Simulation Study of the Arg206Cys Variant in DNASE1L3 Enzyme
Biol. Life Sci. Forum 2022, 20(1), 10; https://doi.org/10.3390/IECBM2022-13378 - 01 Nov 2022
Viewed by 308
Abstract
Genome-wide association studies (GWAS) have identified that one of the autoimmune disease-associated loci, predisposing for the development of Systemic Lupus Erythematosus (SLE), Rheumatoid Arthritis (RA) and Systemic Sclerosis (SSc), is the rs35677470 missense variant of the Deoxyribonuclease I like 3 (DNASE1L3) [...] Read more.
Genome-wide association studies (GWAS) have identified that one of the autoimmune disease-associated loci, predisposing for the development of Systemic Lupus Erythematosus (SLE), Rheumatoid Arthritis (RA) and Systemic Sclerosis (SSc), is the rs35677470 missense variant of the Deoxyribonuclease I like 3 (DNASE1L3) gene, leading to R206C substitution in DNASE1L3 enzyme. Molecular Dynamics (MD) simulations were implemented for the wild type and mutated enzyme, in order to investigate the structure-function relationship that leads to partial loss of enzyme function. Results showed loss of salt bridges, larger fluctuation in active site regions and lower charge of the catalytic and DNA binding sites after the mutation. The present study raises the necessity of in silico methods, in understanding and managing autoimmune diseases. Full article
Abstract
Effects of Glucocorticoid Receptor Activation on the Expression of Intercellular Adhesion Regulatory Genes in Breast Cancer Cells In Vitro
Biol. Life Sci. Forum 2022, 20(1), 11; https://doi.org/10.3390/IECBM2022-13379 - 01 Nov 2022
Viewed by 362
Abstract
Combinations of anticancer chemotherapeutics with glucocorticoids (GCs) are usually used to broaden the therapeutic range of main cytostatic agents and to diminish the side effects of chemotherapy. However, long-term GC administration leads to tumor resistance and the promotion of metastasis. GC effects are [...] Read more.
Combinations of anticancer chemotherapeutics with glucocorticoids (GCs) are usually used to broaden the therapeutic range of main cytostatic agents and to diminish the side effects of chemotherapy. However, long-term GC administration leads to tumor resistance and the promotion of metastasis. GC effects are mediated by the glucocorticoid receptor, which regulates gene expression via DNA-dependent transactivation associated with GC side effects and therapeutically important transrepression. We aimed to determine the molecular markers associated with the GC-stimulated motility and migration of breast cancer cells. We showed that GCs stimulate the invasion and metastasis of breast cancer cells after 120 h of treatment and determined markers of GC-associated adhesion loss. Full article
Abstract
Enlarging the NSAIDs Family: Molecular Docking of Designed Pyrazole and Oxadiazole Derivatives as Novel Anti-Inflammatory Agents
Biol. Life Sci. Forum 2022, 20(1), 14; https://doi.org/10.3390/IECBM2022-13390 - 01 Nov 2022
Viewed by 298
Abstract
The development of the NSAID family has represented a excitingapproach in the treatment of inflammatory disorders, such as arthritis, and for the management of acute pain, in relation to the well-known traditional Non-Steroidal Anti-Inflammatory Drugs (t-NSAIDs). Over the years, research has shown that [...] Read more.
The development of the NSAID family has represented a excitingapproach in the treatment of inflammatory disorders, such as arthritis, and for the management of acute pain, in relation to the well-known traditional Non-Steroidal Anti-Inflammatory Drugs (t-NSAIDs). Over the years, research has shown that essential mediators such as arachidonic acid metabolites are important in inflammation. The cyclooxygenase (COX) and lipoxygenase (LOX) pathways take primary roles in inflammation and are responsible for many human diseases, such as cancer, arthritis, psoriasis, and neurological disorders. Prompted by the pursuit for new cyclooxygenase-2 (COX-2) inhibitors, we have identified novel classes of pyrazole and oxadiazole derivatives as potentially powerful anti-inflammatory molecules. This virtual screening aims to predict the binding affinity of newly designed pyrazole and oxadiazole derivatives against potential molecular targets related to the inflammatory process through the molecular docking approach. Results showed very good anti-inflammatory activity against cyclooxygenase-2 (COX-2) binding protein 1CX2. Additionally, based on the molecular docking results, it was observed that two molecules have good binding affinity with a targeted protein. The issues gained with these classes of compounds represent, currently, a potent stimulus for the further enlargement of the NSAIDs family. Full article
Abstract
Analysis of Lavandula angustifolia Compounds Obtained by Different Extraction Types by GC-MS Technique
Biol. Life Sci. Forum 2022, 20(1), 15; https://doi.org/10.3390/IECBM2022-13374 - 01 Nov 2022
Viewed by 330
Abstract
Lavandula angustifolia is a medicinal plant with important benefits for the human body, exhibiting antimicrobial and antioxidants activities. Scientific data has detailed the fact that lavender extract presented favorable characteristics to health, such as antibacterial, antifungal, antidepressive and anticancer properties. The aim of [...] Read more.
Lavandula angustifolia is a medicinal plant with important benefits for the human body, exhibiting antimicrobial and antioxidants activities. Scientific data has detailed the fact that lavender extract presented favorable characteristics to health, such as antibacterial, antifungal, antidepressive and anticancer properties. The aim of this study was to establish the efficiency of extraction methods by identification and determination of compounds extracted from lavender plant. Different types of extraction were used: ultrasound (50 °C/2 h) and magnetic agitation (ambient temp/24 h) in pure ethanol and hydroalcoholic mixture (ethanol:ultrapure water = 50:50 v/v). GC-MS cromatograph equipment was utilised for detection and quantitative determination of lavender compounds extracted (ex. eucalyptol, linalool, camphor, terpinenol, linalylacetat, etc), an Elite-5MS (5% diphenyl methyl polysiloxane stationary phase) column and a linalool standard. Optimal GC-MS separation parameters were established. In conclusion, it was observed that lavender sample extracted in ethanol, through magnetic agitation at room temperature, represents production by a more efficient method than the others, because more compounds were observed (over 20) than in the other lavender extract samples (approximately 6). Full article
Abstract
Liposomal Supramolecular Strustures Based on the Antiviral Active Nanomaterials
Biol. Life Sci. Forum 2022, 20(1), 18; https://doi.org/10.3390/IECBM2022-13397 - 01 Nov 2022
Viewed by 159
Abstract
Liposomal supramolecular strustures (SMS) are widespread in different areas of modern science [...] Full article
Abstract
Levels of Selected Biochemical Parameters in Homogenates of Isolated Pig Kidneys Flushed with Preservative Solution Supplemented with Selenium and Prolactin
Biol. Life Sci. Forum 2022, 20(1), 19; https://doi.org/10.3390/IECBM2022-13513 - 09 Nov 2022
Viewed by 355
Abstract
Selenium in the human body exhibits strong antioxidant and immunomodulatory properties. This bio-element is found in the active centers of antioxidant enzymes, which participate in the elimination of damage caused by free oxygen radicals. It enters into the composition of some proteins that [...] Read more.
Selenium in the human body exhibits strong antioxidant and immunomodulatory properties. This bio-element is found in the active centers of antioxidant enzymes, which participate in the elimination of damage caused by free oxygen radicals. It enters into the composition of some proteins that build cell membranes, and performs stabilizing functions. Its involvement in catalyzing redox reactions helps reduce oxidative stress and potentially minimize ischemia–reperfusion damage in the kidney during ischemia. The aim of this study was to determine whether supplementation of preservative fluid with selenium in the presence of the antioxidant prolactin affects the levels of selected biochemical indices in homogenates of isolated porcine kidneys. The work is part of a series of our team studies to develop the optimal fluid composition for organ perfusion and preservation. Biolasol preservation fluid was modified by adding Se4+ (1 µg/L) and prolactin (0.1 µg/L). The study was conducted on 30 isolated kidneys of Polish Large White pigs. The kidneys were randomly divided into 3 groups (n = 10 in each) and were washed with preservative fluids: Biolasol control kidneys (C), Biolasol+Se (A1), Biolasol+Se+PRL (A2). After 48 h of preservation and perfusion, kidney sections were excised. Selected biochemical markers were determined in the tissue homogenates: protein and creatinine concentration. The study was performed in accordance with the recommendations of the II Local Ethics Commission for Animal Experiments in Cracow, Poland (number 1046/2013), and in accordance with the European Union Directive (EU guideline 93/119/EC). Supplementation of Biolasol solution with selenium and prolactin caused a statistically significant reduction in protein and creatinine levels compared to the control group in homogenates of isolated pig kidneys. Protein concentrations were: 2.5 ± 0.1 mg/g (group C) vs. 0.9 ± 0.2 mg/g (group A2) (p < 0.05), and creatinine concentrations were: 2.5 ± 0.1 mg/g (group C) vs. 1.9 ± 0.2 mg/g (group A2) (p < 0.05). In contrast, the use of selenium alone (without PRL shielding) resulted in a statistical increase in marker concentrations. Protein concentrations were 20% higher compared to Biolasol (p < 0.05), and creatinine concentrations were 16% higher compared to Biolasol (p < 0.05). Presumably, there was an accumulation of selenium in the organ, exacerbating the resulting damage. Selenium and prolactin added to Biolasol fluid show protective effects on nephrons. Selenium (IV) as a component of Biolasol solution adversely affects renal protection during ischemia. Full article
Abstract
Pipeline to Develop and Characterize a Potential Regenerative Topical Treatment Based on Lavender Essential Oil and the CW49 Peptide
Biol. Life Sci. Forum 2022, 20(1), 20; https://doi.org/10.3390/IECBM2022-13396 - 01 Nov 2022
Viewed by 318
Abstract
Wound healing in adult mammals results in scar formation, which prevents recovering the full functionality of the original skin. This area of dermatology is constantly evolving and especially focuses on aging and the design of recovery treatments for skin burns. Because it has [...] Read more.
Wound healing in adult mammals results in scar formation, which prevents recovering the full functionality of the original skin. This area of dermatology is constantly evolving and especially focuses on aging and the design of recovery treatments for skin burns. Because it has been reported that Lavandula angustifolia essential oil and the CW49 peptide present regenerative, healing, and anti-inflammatory effects, we selected these two natural compounds to formulate a topical treatment with potential regenerative capability. This was accomplished by synthesizing oil-in-water (O/W) emulsions at 10:90% w/w with lavender oil and the CW49 peptide. The formulations were characterized physicochemically and evaluated in terms of biocompatibility, antibacterial activity, and wound-healing potential. The results showed that emulsions exhibited a droplet size of about 1 µm, a marked pseudoplastic behavior and a superior shelf stability of over 9 months. Additionally, they induced 35% hemolysis when compared with the positive control (similar to commercially available controls), induced platelet aggregation, and have a potent antibacterial activity against Staphylococcus aureus (20% of growth inhibition). The wound-healing potential was preliminarily evaluated for the CW49 peptide in a 2D scratch wound model of human keratinocytes, demonstrating an effective concentration for closure of 20 μg/mL. Thus far, we established a pipeline to develop and characterize the regenerative potential of bio-based topical treatments, particularly those based on lavender essential oil and the CW49 peptide. Full article
Abstract
pH-Dependent Specificity of Papain-Like Cysteine Proteases Is Determined by S1 Binding Pocket
Biol. Life Sci. Forum 2022, 20(1), 21; https://doi.org/10.3390/IECBM2022-13385 - 01 Nov 2022
Viewed by 223
Abstract
Papain-like cysteine proteases (PLCPs) are widely expressed enzymes, the main function of which is low-specific-protein turnover in the acidic conditions of lysosomes. Additionally, these proteases provide specific functions in other compartments such as cytosol, nucleus, and extracellular space. The specificity of each protease [...] Read more.
Papain-like cysteine proteases (PLCPs) are widely expressed enzymes, the main function of which is low-specific-protein turnover in the acidic conditions of lysosomes. Additionally, these proteases provide specific functions in other compartments such as cytosol, nucleus, and extracellular space. The specificity of each protease to its substrates mainly depends on the patterns of the amino acids in the binding cleft. This specificity is highly regulated by media conditions and the presence of accessory proteins. In this study, we examined structural aspects, ensuring the pH-dependent substrate specificity of PLCPs. Experiments employing fluorogenic peptide substrates demonstrated that plant PLCPs and human cathepsins possess a pH-dependent specificity for the residue in the P1 position. X-ray crystallographic studies and molecular simulations allowed the overall structure determination of the enzymes to predict residues in the S1 binding pocket, which can form electrostatic contacts with the substrates. Sequence analysis established the variability of these residues among PLCPs. Based on the obtained data, we designed a peptide inhibitor for human cathepsin L and described its inhibitory potential. As a conclusion, we stated that the S1 binding pocket defines specific pH-dependent recognition of substrates by PLCPs, ensuring multiple physiological functions of these proteases. This work was supported by the Russian Science Foundation (grant No. 22-25-00648). Full article
Abstract
Recent Progress in Molecular Recognition Imaging of Protein Systems at the Nanoscale Level
Biol. Life Sci. Forum 2022, 20(1), 22; https://doi.org/10.3390/IECBM2022-13722 - 21 Nov 2022
Viewed by 134
Abstract
Identification of proteins has received considerable attention in recent years due to the increasing interest in resolving individual biomolecules under physiologically relevant conditions [...] Full article
Abstract
Stability and In Vitro Biosafety Study of an Emulsion of Calendula officinalis L. with Potential Application in Treating Skin Burn Wounds
Biol. Life Sci. Forum 2022, 20(1), 23; https://doi.org/10.3390/IECBM2022-13384 - 01 Nov 2022
Viewed by 235
Abstract
Skin burns are injuries of different degrees of complexity (first to third) caused by physical and/or chemical trauma. There is a necessity to improve the rapid retrieval of superficial wounds (first grade) to enhance epithelization, avoiding dehydration, infections, and scar formation. This work [...] Read more.
Skin burns are injuries of different degrees of complexity (first to third) caused by physical and/or chemical trauma. There is a necessity to improve the rapid retrieval of superficial wounds (first grade) to enhance epithelization, avoiding dehydration, infections, and scar formation. This work proposes an oil in water (O/W) emulsion based on 1% of Calendula officinalis L. extract, due to its recognized traditional uses as medicinal plant in wound care, with reported beneficial secondary metabolites such as Carotenoids, Terpenoids, Flavonoids, Coumarins, and Quinones. The rheological characterization of the obtained emulsions indicated superior stability over time (2 months) and a pseudoplastic and semisolid fluid behavior. Furthermore, the emulsions were evaluated biologically in terms of biosafety in vitro with promising results that showed a hemolytic behavior (72.66%) and a moderate platelet aggregation (73.06%) tendency, which is beneficial as it can contribute to enhancing the healing process. Additionally, the emulsions were characterized physicochemically by frequency curves, flow curves of shear stress, and viscosity. Currently, we are conducting a droplet size distribution assay, and conducting 2D wound healing assays in a scratch model over a monolayer of keratinocytes. Thus far, the results hold much promise and indicate that these emulsions can be potentially employed in the treatments of burn wounds. Full article
Abstract
Positive Pharmacological Modulation of Hsp70 in Recovery of Brain Energy Metabolism in Various Models of Cerebral Ischemia
Biol. Life Sci. Forum 2022, 20(1), 24; https://doi.org/10.3390/IECBM2022-13511 - 09 Nov 2022
Viewed by 260
Abstract
The aim of this research was to analyze the neuroprotective action of drugs via an evidence-based approach to analyzing the expression of endogenous neuroprotection factors in various experimental models of cerebral ischemia in Wistar white rats (intracerebral hemorrhage, carotid artery occlusion, prenatal hypoxia) [...] Read more.
The aim of this research was to analyze the neuroprotective action of drugs via an evidence-based approach to analyzing the expression of endogenous neuroprotection factors in various experimental models of cerebral ischemia in Wistar white rats (intracerebral hemorrhage, carotid artery occlusion, prenatal hypoxia) to further substantiate their use in the treatment of CNS damage. The experimental studies were carried out in accordance with the “Regulations on the Use of Animals in Biomedical Research” and the European Convention on the Protection of Animals Used for Scientific and Other Purposes. The experiment was approved by the Bioethics Committee of Zaporizhzhia State Medical University. In studies, it was found that pharmacotherapy of emerging mitochondrial disorders with modulators of HSP70 expression—Cerebrocurin (composition: neuropeptides, S-100 proteins, reelin, factor nerve growth (NGF)), Glutoredoxin, Tamoxifen, (S)-2,6-diaminohexanoic acid, 3-methyl-1,2,4-triazolyl-5-thioacetate significantly (p < 0.05) stimulates energy production in the brain. This is expressed in an increase in the content of a number of intermediates of energy metabolism (glucose, pyruvate, succinate, isocitrate, malate) in the brain tissue, as well as an increase in the activity of pentose phosphate shunt dehydrogenase. A significant (p < 0.05) decrease in the level of lactic acid in the brain indicates a decrease in the proportion of anaerobic glycolysis. The most pronounced energotropic effect was shown by cerebrocurin (150 µL/kg) and (S)-2,6-diaminohexanoic acid 3-methyl-1,2,4-triazolyl-5-thioacetate (50 mg/kg). On the 21st day of treatment of rats with cerebral ischemia, there was a gradual reduction in the severity between animals with simulated pathology and healthy rats, which is associated with actively recovering mitochondrial energy production and cerebral load in general. These changes in energy metabolism correlate with the normalization of the oxidant–antioxidant balance and the concentration of HSP70 and HSF-1a in the cytosol and mitochondria of the cerebral ischemia zone and are manifested in a significant (p < 0.05) decrease in ultrastructural disorders of mitochondria. The energy-tropic and mitoprotective effects of the studied drugs are associated with their ability to protect mitochondrial proteins from oxidative damage by increasing the concentration of HSP70, prolong the lifespan of HSF-1a, and activate alternative energy generation pathways. Full article
Abstract
Natural Diversity of Telomeric DNA Sequences in Bryophytes
Biol. Life Sci. Forum 2022, 20(1), 25; https://doi.org/10.3390/IECBM2022-13740 - 25 Nov 2022
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Abstract
Telomeres are important specialized nucleoprotein structures at the ends of linear eukaryotic chromosomes involved in protecting DNA from damage. The functional characteristic of telomeres is the length. Changes in telomere structure is of enormous evolutionary importance as they can affect whole-genome stability. However, [...] Read more.
Telomeres are important specialized nucleoprotein structures at the ends of linear eukaryotic chromosomes involved in protecting DNA from damage. The functional characteristic of telomeres is the length. Changes in telomere structure is of enormous evolutionary importance as they can affect whole-genome stability. However, plants remain a poorly studied group in light of telomere biology. Bryophytes are a prospective plant group for understanding the protection and evolution mechanisms of plant telomeres. The aim of this work was to determine natural variability of telomere lengths in different bryophyte species and ecotypes. We used axenic cultures of moss Physcomitrium patens ecotypes (Gransden, Reute, Villersexel, Kaskaskia), moss Ceratodon purpureus (male R40 and female GG1), as well as natural isolates Sphagnum fallax MW, S. girgensohnii and Sphagnum sp. The analysis of the telomeres was carried out by the TRF method. It was found that different ecotypes of P. patens plants have different telomere lengths ranging from 1000 to 1500 bp. The mean telomere length in a female plant C. purpureus was 480–500 bp, and in a male plant 900–1000 bp. The telomere lengths of S. fallax MW and S. girgensohnii were mean ~2000 bp. However, telomeres were slightly shorter in isolates of Sphagnum sp., whose telomere length ranged from 1100 to 1500 bp. In addition, it was found that all studied bryophytes had differences in the location of specific telomeric sequences. Thus, we have shown that the telomere lengths of bryophytes can vary both between species and within one species. Furthermore, interstitial telomeric DNA is a distinguishing characteristic of bryophyte genomes. Full article
Abstract
The Validation and Amplification of Random DNA Libraries with Modified Nucleobases for Click-SELEX
Biol. Life Sci. Forum 2022, 20(1), 26; https://doi.org/10.3390/IECBM2022-13692 - 17 Nov 2022
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Abstract
Aptamers are nucleic acid ligands which exhibit specific binding to a desired target. Oligonucleotides are limited in their ability to form hydrophobic interactions. The affinity of aptamers can be improved with the introduction of hydrophobic modifiers into the structure of nitrogenous bases. Click-SELEX [...] Read more.
Aptamers are nucleic acid ligands which exhibit specific binding to a desired target. Oligonucleotides are limited in their ability to form hydrophobic interactions. The affinity of aptamers can be improved with the introduction of hydrophobic modifiers into the structure of nitrogenous bases. Click-SELEX is a powerful tool used for the development of aptamers containing modified nucleobases. This technology assumes the addition of an azide-bearing modifier of choice to the alkyne-modified random DNA library using click chemistry. The synthesis and amplification of modified libraries can limit the performance of click-SELEX. This research aims to validate the modified DNA libraries using NGS and to study specific aspects of their amplification. Commercially available libraries with alkyne-modified uridine (5-ethynyl-deoxyuridine (EdU) instead of thymidine were obtained in four variants with different nucleobase distributions. Alkyne-containing (before clicking of a hydrophobic moiety) and azide-modified (after clicking) libraries were amplified with four different DNA polymerases. All the enzymes screened were able to amplify the non-natural DNA template, and the best amplification efficiency was shown for the Taq DNA polymerase. The NGS of alkyne-containing libraries confirmed the correct length and high diversity of the libraries, as well as the uniformity of nucleotide distribution per position. No significant difference between the sequenced samples amplified with two different DNA polymerases was recognized. In all four libraries, the content of EdU was lower than it was assumed during the chemical synthesis; 14, 13.0, 16.4, and 18.5% EdU values were detected in the random core instead of 20, 20.4, 23.1, and 25% in the theoretical EdU content. To achieve the equimolar distribution of nucleobases, a higher proportion (more than 25%) of EdU should be used during the chemical synthesis of the library. We believe these results can provide an experimental basis for the expansion of click-SELEX technology within routine aptamer research. Full article
Abstract
Formulation and Characterization of a Methacrylated Chitosan Topical Treatment with Dispersed Magnetite Nanoparticles Functionalized with Hydrophobic Drugs Encapsulated in Liposomes
Biol. Life Sci. Forum 2022, 20(1), 27; https://doi.org/10.3390/IECBM2022-13509 - 09 Nov 2022
Viewed by 253
Abstract
Cutaneous administration has advantages over the oral or intravenous route, such as convenience for the patient, avoiding hepatic metabolism, and providing sustained administration of the active component over long periods of time. A major challenge in this route is the administration of drugs [...] Read more.
Cutaneous administration has advantages over the oral or intravenous route, such as convenience for the patient, avoiding hepatic metabolism, and providing sustained administration of the active component over long periods of time. A major challenge in this route is the administration of drugs that are difficult to penetrate. For these, it is necessary to design delivery vehicles that help increase the stability of the active components and facilitate transport across the skin barrier. In this work, magnetoliposomes (MLPs) immobilizing magnetite nanoparticles (MNPs) have been realized. MNPs act as a nanocarrier for hydrophobic drugs, such as doxorubicin (DOX). To facilitate topical application, MLPs were dispersed in photoresponsive methacrylated chitosan hydrogels. For this purpose, the MLPs were synthesized by coprecipitation of FeCl3 and FeCl2. Subsequently, they were silanized and functionalized by a PEG spacer to bind DOX. The success of each functionalization step was evaluated by Fourier transform infrared spectroscopy (FTIR) and thermogravimetric analysis (TGA). The size and morphology of the PEG-DOX-MNPs were analyzed by DLS and TEM. Then, the MNPs-PEG-DOX MNPs were encapsulated in liposomes synthesized by the layer hydration method. Dispersion of MLPs in the hydrogel, followed by crosslinking with visible blue light, was performed. Preliminary FTIR results indicate a correct synthesis and functionalization of the MNPs, as indicated by the presence of bands corresponding to the Si-O stretching vibration at 1029 cm−1 and Fe-O absorption bands around 560 cm−1. TGA results showed a weight loss of 3.5% for MNPs from 200 to 400 °C, which was attributed to silane ligands. The hydrodynamic diameter of the MNPs was 140 nm with polydispersity indices of 0.16. In a future work, DOX will be conjugated to MNPs and MLPs will be synthesized for dispersion in the hydrogel. Subsequently, drug release kinetics tests will be performed under relevant conditions. Full article
Abstract
DNA Obtained by Ab Initio Synthesis Forms Hyperbranched Net-like Structure
Biol. Life Sci. Forum 2022, 20(1), 28; https://doi.org/10.3390/IECBM2022-13691 - 17 Nov 2022
Viewed by 256
Abstract
Ab initio DNA synthesis refers to the unusual synthesis of dsDNA (with a length ranging from tens of bp to kbp) by thermophilic DNA polymerases from free dNTPs in the complete absence of added DNAs. As commonly believed, the reaction product is a [...] Read more.
Ab initio DNA synthesis refers to the unusual synthesis of dsDNA (with a length ranging from tens of bp to kbp) by thermophilic DNA polymerases from free dNTPs in the complete absence of added DNAs. As commonly believed, the reaction product is a linear double-stranded DNA in the B form. However, an extremely low efficiency of cloning and the failure to hydrolyze high-molecular-weight DNA, as well as the presence short repeats, palindromes, and AT-rich repeats in the sequence, mean that a more complex spatial structure of this DNA can be assumed. The AFM coupled with nuclease analysis revealed that high-molecular-weight dsDNA products branched and formed net-like structures. The DNA contained single-stranded and triple-stranded segments. These net-like structures may be assumed to be three-dimensional (3D). The present work was the first detailed investigation of ab initio synthesis products. The results may be useful to develop techniques requiring the synthesis of large amounts of DNA with complex spatial structures. Full article
Abstract
Modeling and Simulation of Magnetoliposome Formation by Encapsulation of Core-Shell, Magnetite-Chitosan Nanoparticles in Liposomes Enabled by a Low-Cost Microfluidic System
Biol. Life Sci. Forum 2022, 20(1), 29; https://doi.org/10.3390/IECBM2022-13398 - 01 Nov 2022
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Abstract
Research in nanostructured materials has led to the development of different applications of relevance in the fields of medicine and biomedical engineering. In this regard, the field of drug delivery has probably benefited the most due to the possibility to engineer vehicles of [...] Read more.
Research in nanostructured materials has led to the development of different applications of relevance in the fields of medicine and biomedical engineering. In this regard, the field of drug delivery has probably benefited the most due to the possibility to engineer vehicles of high potency and increased activity and selectivity toward selected intracellular targets. Such vehicles can therefore potentially address one of the major cornerstones of modern pharmacology, which is increasing the bioavailability of drugs of low permeability. Our research group has developed cell-penetration nanobioconjugates by interfacing several nanomaterials (e.g., chitosan, gelatin nanoparticles, graphene oxide, and magnetite) with translocating peptides. The obtained nanobioconjugates have demonstrated facilitated cell internalization and endosomal escape abilities. To improve cell penetration even further, we encapsulated the magnetite-based nanobioconjugates into liposomes (to form magnetoliposomes) with very appealing results. Our plan is to expand the available nanoplatforms by combining the attributes of magnetite and polymeric nanoparticles through a core-shell system comprised of magnetite (core) and chitosan (shell). The encapsulation process has been successfully accomplished with the aid of passive micromixers with different channel geometries to favor intimate contact between the dispersed phase (nanoparticles) and the continuous phase (phospholipid solution). To model the encapsulation process, we implemented an Eulerian simulation in the software COMSOL Multiphysics® 6.0 (COMSOL Inc, Stockholm, Sweden) where mixing required the Navier-Stokes equations as governing equations of momentum transport, turbulence, eddy viscosity, and damping functions to approximate turbulence using the κ-ε turbulence model near the walls. The simulation was conducted for the different geometries (i.e., SARS, chambers, and serpentine) and for Reynolds numbers ranging from 0.2 to 10 Also, we tested a low Reynolds turbulent model using the κ-ε model given in the Euler-Euler module. The Euler-Euler approach showed that the encapsulation reaches higher encapsulation efficiency (EE%) values compared with the previously implemented mixture model. Our encapsulation results indicate that including the κ-ε turbulence model with a low Reynolds turbulence model near the walls provides a higher agreement between in-silico and experimental approaches. Future work will be dedicated to evaluating the performance of our previously tested magnetophoretic separators with the newly developed encapsulates, to assure sufficient purity for further biocompatibility testing. Full article
Abstract
Study of Spheroids Fusion via Multiphysics Simulations: Feasibility of Applying Permanent Magnetic Field Gradients
Biol. Life Sci. Forum 2022, 20(1), 30; https://doi.org/10.3390/IECBM2022-13386 - 01 Nov 2022
Viewed by 182
Abstract
Cell spheroids represent a scaffold-free route to form cell aggregates through maximizing cell-cell interactions. Spheroids have gained significant attention for the engineering of multilayer tissues as they offer a closer resemblance of physiological conditions observed in vivo, compared with traditional 2D models where [...] Read more.
Cell spheroids represent a scaffold-free route to form cell aggregates through maximizing cell-cell interactions. Spheroids have gained significant attention for the engineering of multilayer tissues as they offer a closer resemblance of physiological conditions observed in vivo, compared with traditional 2D models where cells are grown on flat surfaces. Multilayered tissues are formed by allowing spheroids to interact with themselves within relevant matrices mimicking native conditions of extracellular matrices. However, such conventional fusion methods provide little control over spheroid–spheroid interactions, making their reproducibility very limited. Additionally, such methods are lengthy, which is undesirable from the scalability and economic viewpoints. We propose a methodology to accelerate spheroid fusion by applying magnetic forces externally on spheroids previously magnetized by the internalization of iron oxide nanoparticles. A base mathematical model of spheroids assembly was implemented in COMSOL Multiphysics and involved a laminar two-phase field approach, which was validated by employing a novel segmentation algorithm. The magnetic effect was introduced by an applied volumetric magnetic force, which was generated by the action of two neodymium permanent magnets positioned perpendicular to the computational domain. Our simulations showed that magnetized spheroids fusion can be accelerated by about 55% after applying a magnetic force. In addition, we successfully tested a new modeling approach that allowed taking into account interactions between spheroids and the medium, as evidenced by a standard error of only 13% with respect to the experimental results shown in the literature. Importantly, these simulations also showed that the time required to fuse the spheroids is reduced by about 55%. We are currently validating the model experimentally on extracellular-matrix-derived hydrogels embedded with magnetized spheroids. Future work will be dedicated to calibrating the model with the collected experimental data. Full article
Abstract
Phytochemical Screening and Antioxidant Activity of Trichosanthes cucumerina, Momordica charantia var muricata and Luffa acutangula
Biol. Life Sci. Forum 2022, 20(1), 32; https://doi.org/10.3390/IECBM2022-13694 - 17 Nov 2022
Viewed by 95
Abstract
Background—TC, LA and MCM plants(family: Cucurbitaceae) are widely used in traditional medicine and are important sources of vegetables in the world [...] Full article
Abstract
Genetically Encoded Fluorescent Probes for Imaging of Intracellular Localization and Activity of SARS-CoV-2 Proteins
Biol. Life Sci. Forum 2022, 20(1), 33; https://doi.org/10.3390/IECBM2022-13406 - 01 Nov 2022
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Abstract
Since December 2019, the problem caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has grown into a global threat. The search for new treatment strategies is strongly associated with both fundamental research into the mechanisms of the virus life cycle and the [...] Read more.
Since December 2019, the problem caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has grown into a global threat. The search for new treatment strategies is strongly associated with both fundamental research into the mechanisms of the virus life cycle and the development of new screening platforms for antiviral drug candidates. In this project, we labeled SARS-CoV-2 membrane proteins M, E, and S and studied their localization in mammalian cell compartments using fluorescent microscopy. We tested the N- and C-oriented sensor designs and different fluorescent proteins. Additionally, we successfully visualized the early stages of M protein transport in real time. We found that the M protein localizes in cell lysosomes, which supports the recent hypothesis that β-coronaviruses use lysosomal organelles for egress instead of the traditional Golgi-mediated secretory pathway. Further, we plan to study the interactions between M, E, and S using the FRET method. In addition, we developed several types of FRET-based and translocational sensors to track SARS-CoV-2 PLpro protease and measure its activity in living cells. The preliminary experiments showed the expected increase in donor fluorescence after proteolysis of the PLpro site between the FRET-pair. The results of the current project will provide unique information on the spatial-temporal dynamics and interaction between SARS-CoV-2 membrane proteins during the viral lifecycle. The developed system for the real-time visualization of PLpro activity can potentially serve as a basis for safe cell antiviral drug screening platforms. The proposed strategy for studying viral proteins combines two important properties. Firstly, research has been conducted on human living cells, which is closely approximated to native conditions in contrast to in vitro experiments. Secondly, the experimental system lacks interaction with a functional virus which makes it completely safe for the researcher. Full article
Abstract
Graphene: A New Material for Wound Healing
Biol. Life Sci. Forum 2022, 20(1), 34; https://doi.org/10.3390/IECBM2022-13693 (registering DOI) - 17 Nov 2022
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Abstract
One of the hot topics in medical research involves using new materials for repairing human tissues. Graphene and its derivatives (which will be abbreviated as “graphene” in the following text for the sake of simplicity) happens to be one such novel material and [...] Read more.
One of the hot topics in medical research involves using new materials for repairing human tissues. Graphene and its derivatives (which will be abbreviated as “graphene” in the following text for the sake of simplicity) happens to be one such novel material and has excellent properties due to its ability to help regenerate tissues. This could be potentially useful for tissue engineering, and skin/muscle/nerve/bone/cartilage repair. Wound healing in eczema, bed sores, or burning accidents have a high risk of bacterial infection, and some graphene materials have shown to provide good therapeutic efficacy in the improvement of wound healing with new dressing materials. The mechanism of wound healing might be due to its anti-bacterial properties, immunomodulatory effects, anti-inflammatory effects, as well as angiogenic properties. The reason behind these unique properties could be graphene’s lack of cytotoxicity, its compatibility with biological material in terms of adhesiveness, and its lack of inhibition of healthy cell migration. Graphene could also have some antibacterial properties which might be due to its dehydrating properties, or the ability of some functional groups to generate free radicals that kill pathogenic bacteria. The biocompatibility could be demonstrated by examination of the adhesion of fibroblast cell lines and the morphology of their filopodia (or the feet). While many model systems concentrate on skin repair and wound healing, the use of graphene is not limited to skin cells. For instance, a human stem cell model has also been used to mimic cell regeneration from acute myocardial infarction (MI) using graphene in the form of hydrogels. This model has established an enhanced cell survival rate, an increased expression of pro-inflammatory factors that aid in the formation of new blood vessels, and early cardiogenic biomarkers that use graphene quantum dots as a soft injectable hydrogel for heart regenerative function after MI. The study of graphene and graphene-based materials on inflammatory biomarkers and acute phase proteins will be the subject of investigation in this review. Full article

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Proceeding Paper
Bivalent System of Deoxyribozymes for Efficient RNA Cleavage
Biol. Life Sci. Forum 2022, 20(1), 6; https://doi.org/10.3390/IECBM2022-13510 - 09 Nov 2022
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Abstract
The goal of gene therapy is to treat diseases through gene editing and modification of gene expression. Viral vectors, antisense oligonucleotides (ASO) and RNA interference (RNAi) are some of the gene therapies under development. DNAzymes, or deoxyribozymes (Dzs), are simple in design tools [...] Read more.
The goal of gene therapy is to treat diseases through gene editing and modification of gene expression. Viral vectors, antisense oligonucleotides (ASO) and RNA interference (RNAi) are some of the gene therapies under development. DNAzymes, or deoxyribozymes (Dzs), are simple in design tools for RNA-cleaving and oligonucleotide-based gene therapy. However, Dz affinity and efficiency remain a significant challenge to be addressed. In nature, multiple ligands bind to targets at multiple sites with high affinity and avidity, resulting in effective inhibition or stimulation of biological responses. Scientists have reported multivalency in biological systems as a powerful strategy for achieving high-affinity molecular recognition. Our main goal was to improve the affinity and efficiency of DNAzyme in RNA cleaving by developing and optimizing 10-23 Dz base gene therapy molecules. We used 10-23 Dz to target folded mRNA and tested their knockdown efficiency in an in vitro physiological buffer. Next, we optimized 10-23 Dz by designing monovalent Dzs with varied arm lengths (short arms with 1–2 nucleotides less and long arms with 1–2 nucleotides more) to find the most efficient and stable construction and tested their efficiency in a cooperative association. It was observed that cooperative Dz1-Dz2 association was 3 times more efficient in comparison with separately working Dz1 and Dz2. Further investigation found that different lengths (−/+1, +2 nucleotides) of binding arms influenced the efficiency of 10-23 Dz. The result indicated that, as the length of the binding arms increased, the efficiency also increased. This research demonstrated that multivalent associations of DNAzymes have great potential to increase DNAzyme-cleaving RNA affinity and efficiency as a therapeutic agent. Full article
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Proceeding Paper
A Mini Literature Review on Current Advancements in Protein Purification Techniques
Biol. Life Sci. Forum 2022, 20(1), 12; https://doi.org/10.3390/IECBM2022-13507 - 09 Nov 2022
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Abstract
Protein purification is an ever-vital technique for academia and industry. This paper mainly reviews and discusses one of the core components of proteomics—the latest advances in separation technology for protein components—focusing on five different methods. The multi-column plate adapter (MCPA) system is incredibly [...] Read more.
Protein purification is an ever-vital technique for academia and industry. This paper mainly reviews and discusses one of the core components of proteomics—the latest advances in separation technology for protein components—focusing on five different methods. The multi-column plate adapter (MCPA) system is incredibly economical for protein treatment research to purify samples. Because of the affinity of excess Ulp1 protease for SUMO fusion, excess protein products may be obtained within half an hour using this method. Magnetic separation strategies can offer a better protein purification process in the future because of a few advantages. The evaluation established that the aqueous two-phase system (ATPS) technique is a cost-effective, time-saving (30 min), and high-recuperation approach that can be scaled up for commercial purposes. Therefore, the ATPS may be a viable single-step separation purification method, moving away from multi-step purification such as the chromatography technique. Our review provides a technique capable of efficient protein purification. Full article
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Proceeding Paper
Determining the Effect of Plant Extracts on the Development and Characterization of Biodegradable Composite Films from Corypha umbraculifera L. Stem Starch
Biol. Life Sci. Forum 2022, 20(1), 13; https://doi.org/10.3390/IECBM2022-13393 - 01 Nov 2022
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Abstract
Films and coatings fabricated with renewable biopolymers and antimicrobial agents have attracted research interest owing to their contribution to food safety and biodegradability. The study aimed to determine the effect of natural plant extracts from the leaves of curry tree, neem, tulsi, and [...] Read more.
Films and coatings fabricated with renewable biopolymers and antimicrobial agents have attracted research interest owing to their contribution to food safety and biodegradability. The study aimed to determine the effect of natural plant extracts from the leaves of curry tree, neem, tulsi, and Mexican mint in developing and characterizing biodegradable composite films of talipot starch and carboxymethyl cellulose (CMC) matrices. Talipot starch isolated from the stem pith of talipot palm (Corypha umbraculifera L.) is an underutilized source of starch with a high yield (76%). All composite films were prepared using the solution blending-casting method. The dominant properties of biodegradable films such as structural, morphological, barrier, and antimicrobial properties were studied. The relative crystallinity (RC) of composite films comparatively decreased with native talipot starch film. The surface of the talipot starch film made with CMC and plant extracts showed higher roughness and opacity. Incorporation of plant extracts into talipot starch and CMC matrices decreased water vapor permeability (WVP) and oxygen permeability (OP), indicating the improved barrier properties of the films. Antimicrobial activity as assessed by the inhibition zone method showed that composite films exhibited excellent antimicrobial activity against Staphylococcus aureus and Escherichia coli. These results revealed that biodegradable composite films from the non-conventional starch of talipot palm can possibly be used as a substitute for the one- time use petroleum-based films and can be used as a bioactive packaging material for food applications. Full article
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Proceeding Paper
Impact of Different Cross-Linking Agents on Functional, Rheological, and Structural Properties of Talipot Palm Starch: A New Source of Stem Starch
Biol. Life Sci. Forum 2022, 20(1), 16; https://doi.org/10.3390/IECBM2022-13391 - 01 Nov 2022
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Abstract
The talipot palm (Corypha umbraculifera L.) is an unconventional source of stem starch, with a 76% starch yield. Talipot starch was cross-linked with epichlorohydrin (EPS) and phosphoric acid (PS), which significantly altered the functional, pasting, rheological, and structural properties. Amylose content of [...] Read more.
The talipot palm (Corypha umbraculifera L.) is an unconventional source of stem starch, with a 76% starch yield. Talipot starch was cross-linked with epichlorohydrin (EPS) and phosphoric acid (PS), which significantly altered the functional, pasting, rheological, and structural properties. Amylose content of talipot starch was significantly decreased and relative crystallinity was increased by cross-linking. The EPS cross-linked talipot starch has a higher swelling capacity and PS cross-linked talipot starch has a lower swelling capacity. A similar trend was observed in the pasting profile, and EPS has higher peak and final viscosities. The native starch gel has a hardness of 45.54 N, which was increased to 149.69 N in EPS, whereas it decreased to 13.62 N in PS. The paste clarity of all the samples was found to be decreased during cold storage. As compared to the native, both EPS and PS exhibited higher paste clarity. The percent syneresis was considerably decreased in cross-linked starch (CLS). The magnitude of both G′ and G″ was significantly changed by cross-linking. The EPS exhibited increased G′ and G″ values, whereas these decreased in PS. Full article
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Proceeding Paper
The Effect of Heat-Moisture Treatment (HMT) on the Structural, Functional Properties and Digestibility of Citric Acid-Modified Plectranthus rotundifolius (Hausa Potato) Starch
Biol. Life Sci. Forum 2022, 20(1), 17; https://doi.org/10.3390/IECBM2022-13392 - 01 Nov 2022
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Abstract
In this study, Hausa potato starch was subjected to single and dual modifications by HMT and citric acid. The parameters such as relative crystallinity, pasting, functional properties, and in vitro digestibility were studied. The XRD analysis showed an A-type diffraction pattern for the [...] Read more.
In this study, Hausa potato starch was subjected to single and dual modifications by HMT and citric acid. The parameters such as relative crystallinity, pasting, functional properties, and in vitro digestibility were studied. The XRD analysis showed an A-type diffraction pattern for the native starch and was unaffected by all modifications. The relative crystallinity and the gelatinization enthalpy significantly decreased (p ≤ 0.05) on dual modification. A new peak at 1724 cm−1 was observed in the FT-IR spectra of citric acid-modified starch. The peak intensity became stronger in HMT followed by citric acid-modified starch, which was consistent with the results of degree of substitution. The native starch showed a peak viscosity of 3343 cP and significantly decreased on citric acid modification, and HMT treatment increased the effectiveness of the citric acid modification. The in vitro digestibility of Hausa potato starch was significantly affected by both single and dual modifications. The increased DS and RS content of the dual-modified starch suggests that the HMT served as a pre-treatment and favored the production of the citrate starch. The HMT-citric acid dual modification method was shown to modify the starch properties and could be used as a substitute for producing low glycaemic index foods. Full article
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Proceeding Paper
In Silico Determination of Changes in Transcription Factor Binding Sites for the Preeclampsia Risk Haplotype in the Regulatory Region of the FLT1 Gene
Biol. Life Sci. Forum 2022, 20(1), 31; https://doi.org/10.3390/IECBM2022-13721 - 18 Nov 2022
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Abstract
Preeclampsia (PE) is one of the most common complications of pregnancy that occurs in 3–8% of pregnant women, being one of the top five causes of maternal morbidity and mortality. We found that PE-associated polymorphisms near the FLT1 gene are located in the [...] Read more.
Preeclampsia (PE) is one of the most common complications of pregnancy that occurs in 3–8% of pregnant women, being one of the top five causes of maternal morbidity and mortality. We found that PE-associated polymorphisms near the FLT1 gene are located in the same regulatory region. In combination, these polymorphisms can form a genetic pattern which affects the development of the pathology by forming a PE risk haplotype. When analyzing the changes in TPFS, which are characteristic only for the risk haplotype with a prevalence in the European population of 0.0825, we found that five TFBs change. The number of TFBSs for ELF1 and SPIB increases, whereas the number of TFBSs for POLR2A and KLF15 decreases (not expressed in the placenta).The newly emerged transcription factor binding site KAT5 acquires a promoter signature in the placenta only after 118 days of pregnancy, whereas before the 118th day, only the DNase signature is observed. Theoretically, the appearance of a new TFBS can increase the expression of FLT1, causing an imbalance of angiogenic/antiangiogenic factors, which is characteristic of PE. Full article
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