Polyacrylamide Gel Research Articles

Effect of green tea or black tea extract on lipid and protein oxidation in Cantonese sausage.

Natural polyphenols offer a safer alternative to synthetic antioxidants in meat products. This study investigated the efficacy of green tea and black tea extracts as natural antioxidants in Cantonese sausages to inhibit lipid and protein oxidation. Sausages were prepared with the addition of different concentrations - 100, 300, and 600 mg kg-1 total polyphenols (TP) - of green tea or black tea extract. Oxidation of the sausages was assessed through thiobarbituric acid reactants, carbonyl content, and thiol content, whereas consumer acceptability was evaluated based on texture, color, and sensory analysis. The tea extracts inhibited malondialdehyde production and reduced the thiobarbituric acid reactive substance value from 23.72 mmol MDA g-1 to less than 1.94 mmol MDA g-1. However, the addition of tea extracts decreased the thiol content and caused the loss of myosin heavy chain and actin bonds in sodium dodecyl sulfate polyacrylamide gel electrophoresis. Although the addition of tea extracts increased the redness and hardness of the sausage, no significant difference in consumer acceptance between the control and treatment groups was observed in the sensory analysis. The tea extract inhibited the oxidation of lipids in Cantonese sausage. There was no negative effect on the sensory characteristics of sausages. The use of tea extracts as natural antioxidants in Cantonese sausage is therefore feasible. © 2024 Society of Chemical Industry.

Producción de esterasa por Fusarium culmorum crecido en presencia de diferentes concentraciones di(2-etilhexil) ftalato y de ion calcio en fermentación sumergida

Phthalates are plastic additives used as plasticizers in the manufacture of flexible plastic materials. These compounds, such as di(2-ethylhexyl) phthalate (DEHP) are toxic substances to the environment and human health, and have been detected as environmental pollutants. In this study, the activity of esterase produced by Fusarium culmorum grown in media supplemented with different concentrations of DEHP (1.5, 3 and 4 g/L) and calcium ion (0.1 and 0.5 g/L) in submerged fermentation was characterized by biochemical tests and polyacrylamide gel electrophoresis. The greatest esterase activity was observed in cultures grown in media added with 3 g of DEHP/L and 0.5 g of calcium ion/L, followed by those cultures grown in media containing 1.5 g of DEHP/L. Whereas cultures grown on 4 g of DEHP/L had the lowest esterase activity. In general, cultures added with 0.5 g of calcium ion/L had the maximum enzymatic activity and the highest enzymatic productivity in the media tested. High concentration of DEHP were used as the sole source of carbon and energy by F. culmorum. It is suggested that the calcium ion favored the esterase production during the DEHP biodegradation. However, further studies are needed to understand the relationship between calcium ion, fungal growth, and enzyme production to provide the appropriate concentration of this ion in the culture medium.

Effects of spray drying and freeze drying on the protein profile of whey protein concentrate.

Whey protein concentrate (WPC) is consumed for its high protein content. The structure and biological functionality of whey proteins in WPC powders may be affected by the drying technique applied. However, the specific impact of spray drying and freeze drying on the overall protein profile of whey protein derived from sweet whey streams at scale is unknown. Herein, we examine the effects of commercial-scale freeze drying and spray drying on WPC to determine which method better preserves bioactive whey proteins, with the goal of helping the dairy industry create high-value products that meet the growing consumer demand for functional dairy products. WPCs were produced from pasteurized liquid whey using either a commercial spray dryer or freeze dryer. A variety of analytical techniques, including enzyme-linked immunosorbent assay, polyacrylamide gel electrophoresis, and bottom-up proteomics using liquid chromatography-tandem mass spectroscopy were used to identify, quantify, and compare the retention of bioactive proteins in WPC before and after spray drying and freeze drying. In addition, the extent of denaturation was studied via solubility testing, differential scanning calorimetry, and hydrophobicity assessment. There was little to no difference in the retention or denaturation of key bioactive proteins between spray-dried and freeze-dried WPC powders. There was a higher percentage of select Maillard modifications in freeze-dried and spray-dried powders than in the control. The lack of significant differences between spray drying and freeze drying identified herein indicates that freeze drying does not meaningfully improve retention of bioactive proteins compared with spray drying when performed after multiple pasteurization steps. PRACTICAL APPLICATION: This study aimed to provide insight into the impacts of spray drying versus freeze drying on whey proteins. Overall, our results indicate that for commercial dairy processing that involves multiple rounds of pasteurization, freeze drying does not meaningfully improve the retention of bioactive proteins compared with spray drying. These findings may help the food and dairy industry make informed decisions regarding the processing of its whey protein products to optimize nutritional value.

Biotechnological methods for separation of pigs pancreas glands protein substances with membrane technologies

The pancreas gland (PG) is a secondary product of livestock processing; it contains a wide range of biologically active compounds. The purpose of this article is to analyze the efficiency of technological approaches for pancreas gland extraction with the help of trehalose and a glycine-proline mixture aimed for recovery and separation of the gland’s protein-peptide compounds. The extraction was conducted with 0.9% NaCl, 0.9% NaCl, with addition of 0.5 M trehalose (0.9% NaCl-0.5 M trehalose) and 0.9% NaCl with addition of 1% glycine and 0.1 M L-proline (0.9% NaCl-1% Gly-0.1M Pro), the ratio of pancreas gland to extractant was equal to 1:5. The concentration of the protein in the supernatants after their extraction was measured by the biuret reaction in a semi-automatic biochemical analyzer Biochem SA. The proteomic composition of the extracts and the native pancreas gland was assessed by one-dimensional Laemmli electrophoresis in a 12.5% polyacrylamide gel and by two-dimensional O’Farrell electrophoresis. When determining the intensity of the protein fractions, it was noted that the methodology of separation of protein-peptide mixtures extracted from the pigs pancreas gland with the extractant 0.9% NaCl-1% Gly-0.1M Pro, ensured the higher extraction of the proteins in comparison with the method of 0.9% NaCl-0.5 M trehalose. Notwithstanding the fact that application of amino acids (glycine and proline) mixture provided for a greater yield of proteins from the extract into the diafiltrate, the experiments in vitro showed that the diafiltrate obtained though trehalose featured higher activity. This may be explained by the fact that after dialysis removal of trehalose from the protein fraction with a molecular weight of less than 50 kDa, its residual quantities were still sufficient to prevent proteins aggregation and, as a consequence, the biological activity of the extracted proteins was preserved, while in the diafiltrate obtained through amino acids mixture where numerous protein aggregates were detected by 2-DE. This study allowed testing the biotechnological methodics on pig pancreatic tissues aimed to intensifying the extraction and separation of protein compounds. The results of the study are important for development of methodological approaches to obtaining the targeted substances for their further utilizing for various purposes.

Tetraspanin 5 orchestrates resilience to salt stress through the regulation of ion and reactive oxygen species homeostasis in rice.

Tetraspanins (TETs) are integral membrane proteins, characterized by four transmembrane domains and a unique signature motif in their large extracellular loop. They form dynamic supramolecular complexes called tetraspanin-enriched microdomains (TEMs), through interactions with partner proteins. In plants, TETs are involved in development, reproduction and immune responses, but their role in defining abiotic stress responses is largely underexplored. We focused on OsTET5, which is differentially expressed under various abiotic stresses and localizes to both plasma membrane and endoplasmic reticulum. Using overexpression and underexpression transgenic lines we demonstrate that OsTET5 contributes to salinity and drought stress tolerance in rice. OsTET5 can interact with itself in yeast, suggesting homomer formation. Immunoblotting of native PAGE of microsomal fraction enriched from OsTET5-Myc transgenic rice lines revealed multimeric complexes containing OsTET5, suggesting the potential formation of TEM complexes. Transcriptome analysis, coupled with quantitative PCR-based validation, of OsTET5-altered transgenic lines unveiled the differential expression patterns of several stress-responsive genes, as well as those coding for transporters under salt stress. Notably, OsTET5 plays a crucial role in maintaining the ionic equilibrium during salinity stress, particularly by preserving an elevated potassium-to-sodium (K+/Na+) ratio. OsTET5 also regulates reactive oxygen species homeostasis, primarily by modulating the gene expression and activities of antioxidant pathway enzymes and proline accumulation. Our comprehensive investigation underscores the multifaceted role of OsTET5 in rice, accentuating its significance in developmental processes and abiotic stress tolerance. These findings open new avenues for potential strategies aimed at enhancing stress resilience and making valuable contributions to global food security.

The influence of polyacrylamide gel substrate elasticity on primary cultures of rat retinal ganglion cells

In vitro primary cell culture models of retinal ganglion cells (RGC) are widely used to study pathomechanisms of diseases such as glaucoma. The biomechanic interaction with the culture substrate is known to influence core cellular functions. RGC cultures, however, are usually grown on rigid plastic or glass substrates. We hypothesized that soft polyacrylamide gel substrates may alter survival and neurite outgrowth of primary cultured RGC.Primary retinal cultures from postnatal (day 1-6) Wistar rats were grown on glass coverslips or polyacrylamide (PA) gel substrate with different Young’s elastic moduli (0.75, 10 or 30 kPa). Substrates were coated with Poly-l-lysine and / or laminin. RGC were immunostained with anti-beta-III-tubulin. Total neurite length, growth cone morphology, RGC density, mitochondrial morphology and transport as well as pro-survival pathways (Erk1/2, Akt, CREB) were assessed.PA gel substrates of E = 10 kPa significantly increased the total neurite length by factor 1.5 compared to glass (p = 0.02). The growth cone area was significantly larger by factor 5.3 on 30 kPa gels (p = 0.01). The presence of a substrate coating was more important for neurite outgrowth and RGC survival on PA gels (poly-l-lysine > laminin) than on glass. Neither mitochondrial morphology and motility nor the activation of pro-survival pathways significantly differed between the four substrates.PA gel substrates significantly enhanced RGC neurite outgrowth. The signaling cascades mediating this effect remain to be determined.

Study on the Recombinant Human Interferon α1b, α2b, and Gamma Transient Expression and in Vitro Activities in Tobacco.

Interferons (IFNs) are universally acknowledged for their pivotal role in antiviral and anticancer responses. Thus, the primary aim of our study was to explore the expressions of IFN-α1b, α2b, and gamma in tobacco leaves via agrobacterium-mediated transient transformation and investigate their possible activities. Briefly, fusion with green fluorescent protein tags aided in detecting the expressed IFN proteins in the foliar tissues. The genetic constructs encoding these fusion proteins were inserted into the MagnICON plant transient expression vector, followed by transformation into the Agrobacterium strain GV3101. The transformed bacteria were then used to infiltrate tobacco leaves. After post-infiltration, protein expression was confirmed within 72 h via sodium dodecyl sulfate polyacrylamide gel electrophoresis, and the fusion proteins were subsequently purified using high-performance liquid chromatography for identification. Both the antiviral and anticancer potencies of these IFN fusion proteins were evaluated using the WISH/VSV (WISH cells/Vesicular stomatitis virus) microneutralization and MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assays, respectively. Results indicated robust expression of the targeted IFN genes in plant tissues and significant biological activities against pathogens and cancer cells. Consequently, this study substantiated the viability of producing these therapeutic proteins in plants, potentially revolutionizing the manufacture of interferons biologically.

Proteomic Analysis of Storage Proteins in Phaseolus vulgaris Associated with Resistance to Water Stress

The common bean (Phaseolus vulgaris), a widely consumed legume in Ecuador, boasts low economic value, significant nutritional contributions, and a remarkable capacity to enhance soil fertility. Despite these attributes, its field productivity often needs to improve, with the water deficit emerging as a primary hindrance. Consequently, genetic enhancements have been incorporated into select varieties, conferring tolerance to specific levels of water scarcity stress. This study aimed to elucidate the distinctions in protein expression patterns responding to water deficiency stress across nine bean varieties. Protein patterns were scrutinized through two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), and selected protein spots were subjected to mass spectrometry analysis (MALDI-TOF MS/RP-LC-MS/MS). A comprehensive identification of 111 proteins was achieved and categorized based on their respective functions. Noteworthy among these were the desiccation-protectant protein (LEA14) and Desiccation-related protein PCC13-62, identified as proteins associated with the response to abiotic stress, particularly prevalent in the INIAP_473 cultivar. These findings underscore the potential for targeted genetic improvements to mitigate the impact of water deficit stress on common bean cultivation, contributing to enhanced agricultural resilience and productivity.

Biochar coupling induced charge transfer switching of a NiAl2O4/NiCr2O4 photocatalyst to enhance photocatalytic degradation of tetracycline hydrochloride

A series of novel (NiAl2O4 (NAO)/NiCr2O4 (NCO)) (S1)/GL heterojunction photocatalysts, denoting as x%GL/S1 (x = 5, 10, 15 and 20), were prepared by the polyacrylamide gel method combined with low temperature sintering technique. Various characterizations confirm that there is special interfacial contact and charge transfer between NAO, NCO and GL. The 10%GL/S1 heterojunction exhibited the best photocatalytic activity and cyclic stability for the degradation of tetracycline hydrochloride (TC). Compared to NAO, NCO and NAO/NCO, the degradation efficiency of 10%GL/S1 was significantly improved, and the degradation percentage reached 83.2% within 30 min. The activity of 10%GL/S1 was studied in relation to catalyst content, drug concentration, pH value, water quality, and co-existing anion. Trapping experiments and free radical verification experiments confirmed that the holes, hydroxyl radicals and superoxide radicals played crucial roles in the photocatalysis process. Possible pathways for TC degradation were studied by HPLC-MS, and the ecotoxicity of TC and its degradation intermediates was evaluated by T.E.S.T. software. The charge transfer and photocatalytic mechanisms confirm that the introduction of GL enhances the interfacial interaction between the components, promotes the carrier migration efficiency, provides a large number of active sites for 10%GL/S1 heterojunction, and produces active free radicals after redox reaction with photoinduced electrons, effectively improving the degradation efficiency of TC.

Binding mechanism and structural characteristics of alloyed protein complex for enhanced solubility of hemp protein isolate

Despite the numerous health benefits and high digestibility of hemp protein isolate (HPI), its low solubility at neutral pH limits its utilization in the food industry. Therefore, we subjected insoluble HPI and soluble mung bean protein isolate (MBPI) to pH co-shifting under extremely alkaline conditions to form an alloyed protein complex (A-HM). At a mass ratio of HPI:MBPI of 50:50, A-HM exhibited the highest solubility (95.30 ± 0.99 %), and also had high resistance to heat treatment. Native PAGE demonstrated the formation of alloyed protein complexes, and particle size analysis revealed that A-HM exhibited small particle sizes and dispersion in water without aggregation of HPI. Owing to their small size, numerous hydrophobic residues and aromatic ring of HPI were exposed on the surface. Hydrophobic interactions predominantly governed the binding force involved in the formation of A-HM. Our findings may enhance HPI applications in the food industry, particularly in plant-based beverages.

Antioxidant analysis of protein from fresh and dry leaf of Orthosiphon aristatus (Blume) Miq.

Orthosiphon aristatus (Blume) Miq. is a folklore plant consumed as brewed tea for various health benefits. The white variety of the plants leaf and stem are predominantly sold in the market in dried form, blooming as a natural herbal product. To date, no proteomics and antioxidant studies are available on the fresh and dried leaf protein extract of O. aristatus since most studies take an interest in the crude extracts of plants. Thereupon, this study focuses on the One-dimensional (1D) electrophoretic pattern of the fresh and dry leaf determined via sodium dodecyl-sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Additionally, antioxidant proteins are currently being focused on for their link to controlling disease through their potential to destroy free radicals that are present in excess. Hence, an antioxidant assay was conducted for the fresh and dry leaf protein extract using 2,2-Diphenyl-1-picrylhydrazyl (DPPH) free radical assay and ferric reducing antioxidant power (FRAP) assay. The electrophoretic pattern obtained resulted in approximately 16 and 10 bands for the fresh and dry leaves respectively. As for the antioxidant activity, the dry leaf had higher radical scavenging activity and higher reducing power compared to the fresh leaf. The findings obtained provides insight in to the protein content and antioxidant activity found in the protein extract of fresh and dry leaves of O. aristatus. Hence, further detailed study of the protein extracts could further unveil their therapeutic potential.

Rhizosphere bacterial community influence on <i>Deschampsia antarctica</i> È Desv. adaptability in context of temperature near plants in local spatial scales of Galindez Island, Argentine Islands (the maritime Antarctic)

The effect of rhizosphere bacterial community index (Irbi) influence on the nine populations of Antarctic hair grass (Deschampsia antarctica) adaptability was studied in the Galindez Island (summer season 2017/18). Moreover, the corresponding influence indices Irbi (i=1÷9) and Irbpi (p=1÷5 for the most common bacteria) were evaluated as well. The objective was to compare the Irbi and Irbpi series with the united temperature influence index on plant populations (It1i(z)) series and the united quality latent index of adaptability Iq1i. The study used data on the rhizosphere metagenome composition based on 16S RNA analysis. Methods determining the plant number in populations, and measuring the morphometric indices of D. antarctica populations were used. Reserve and protective seed proteins spectra were studied by polyacrylamide gel electrophoresis. Method of extreme grouping the spatial variables of these indices was applied for nine populations to obtain a Iq1I and It1i and Irbi, Irbpi. Sets of united indices were compared by regression technique. A comparative statistical analysis of the It1i and Irbi, Irbpi sets in this season was carried out. This possible influence appeared to be individual for each D. antarctica studied population. In each population, part of the plants reacted positively to the bacteria influence, while the other part either did not react or reacted negatively. Dependence of the plant adaptation indices on rhizospheric bacterial communities z(x) is shown in our data. This means that the rhizosphere bacterial community and temperature-dependent rhizosphere bacteria (x) can play an active role in plant adaptation of D. antarctica populations (z) to individual temperature conditions in the microscale of Galindez Island from a biological point of view.

Selection of antibacterial aptamers against Pseudomonas plecoglossicida and analysis on their potential binding proteins

Pseudomonas plecoglossicida is one of the main pathogens causing visceral white spot disease of the large yellow croaker (Pseudosciaena crocea). Although antibiotics are used to control P. plecoglossicida infections, the long-term and large-scale use of antibiotics can lead to the development of drug-resistant bacteria as well as environmental pollution. Therefore, it is necessary to develop novel antibacterial agents to treat P. plecoglossicida infections. In this study, we first developed a two-step-centrifugation method to isolate live and dead P. plecoglossicida cells. Subsequently, we used the Systematic Evolution of Ligands by Exponential Enrichment (SELEX) screening based on inhibition rate to directly isolate antibacterial aptamers from the random library without post-processing. We isolated five antibacterial aptamers, namely B1, B2, B4, B8, and B109, which showed good inhibitory effects against P. plecoglossicida. Among these, B4 showed the highest inhibitory effect (62.40 ± 4.17 %). Further analysis revealed no positive correlation between the inhibitory effect of aptamers and their affinities with the target bacterium. The B4- and B109-binding proteins were isolated from P. plecoglossicida by magnetic separation and sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE). Mass spectrometry analysis revealed that the 26 kDa ribosomal protein L2 was the probable B4-binding protein, while the 26 kDa ribosomal protein S3 and 75 kDa ribosomal protein S1 or succinate dehydrogenase flavoprotein subunit were the probable B109-binding proteins. Structural and subcellular localization analyses of these potential B4- and B109-binding proteins were also conducted. Our findings suggest that the inhibitory activity of the antibacterial aptamers against P. plecoglossicida may be mediated via their interaction with the ribosomal proteins, which can interfere with the protein synthesis process in the bacterium, affecting its growth. These findings provide the scientific basis for the development of functional antibacterial aptamers and the elucidation of their mechanisms of action.

Effect of pH-shifting and ultrasound on soy/potato protein structure and gelation

The substitution of meat proteins with plant-based proteins from various sources is often motivated by nutritional considerations. However, the inherent limited solubility of plant proteins, which results in suboptimal techno-functional properties, remains a persistent challenge in food formulation. The purpose of this study was to investigate the impact of pH-shifting (P), high-intensity ultrasound (U), and ultrasound-assisted pH-shifting on the structure and solubility of soy protein isolate (SPI), potato protein isolate (PPI), and the soy/potato protein complex (SPI/PPI complex). Additionally, a detailed examination of the gel properties of SPI, PPI, and the SPI/PPI complex was conducted to assess the changes that occurred before and after modification. The results of particle size, zeta potential, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), free sulfhydryl (R-SH), surface hydrophobicity (H0), Fourier transform infrared spectrum (FTIR) and fluorescence intensity showed that P and U treatment promoted the unfolding of proteins structures and decreased particle size, and augmented protein-water interactions, leading to enhanced solubility. The pH-shift and ultrasound (PU) treatment further amplified these structural alterations in proteins. Consequently, the gelling characteristics of the modified proteins were substantially improved, with notable increases in hardness, elasticity, and water-holding capacity (WHC). The findings facilitate developing more targeted plant protein-based gel products that specific techno-functional attributes, concurrently allowing for modifications to their nutritional profiles.

Cereal barcoding by assessing intron length polymorphism of γ-tubulin genes

Aim. The aim of the study was to obtain typical DNA profiles for common cereal plants using a method of assessing γ-tubulin intron length polymorphism and to evaluate the possibility of using this gene for plant species barcoding. Methods. Cereals, in particular wheat, aegilops, barley, and rice, have been studied. The method of γ-tubulin intron length polymorphism assessment was used. Amplified DNA fragments were separated via polyacrylamide gel electrophoresis under non-denaturating conditions and were visualized via silver nitrate staining. Results. The analyzed cereal species-specific DNA profiles of amplicons of the γ-tubulin gene introns were obtained, which made it possible to differentiate the species. Fingerprinting data was used for cluster analysis and dendrogram construction. Conclusions. The feasibility of using the proposed cereal barcoding method has been established. The studied plant DNA profiles can be effectively used in assessing the quality of food products such as flour.

Identification of molecular markers linked to leaf rust resistance gene Lr13 in wheat

Aim. Identification of codominant molecular markers closely linked to the Lr13 leaf rust resistance gene. Methods. PCR with detection in a polyacrylamide gel. Hybridological analysis. Phytopathology evaluation. Statistical analysis. Results. PCR analysis of varieties carrying the Lr13 gene and near-isogenic lines of the Thatcher variety with genes for resistance to leaf rust Lr13 (TcLr13), Lr16 (TcLr16), Lr23 (TcLr23) and the F2 population TcLr13 x TcLr34 by the microsatellite loci Xbarc13, Xbarc55, Xgwm148, Xwmc261, Xgwm271, Xwmc272, Xwmc344, Xbarc361, Xgwm410, Xwmc474, Xwmc477 and Xgwm630 was performed. To test the two microsatellite loci Xgwm148 and Xwmc344 as candidate markers for the Lr13 gene, we used the F2 population, which was obtained from crossing two near-isogenic lines of the Thatcher variety with leaf rust resistance genes Lr13 (TcLr13) and Lr34 (TcLr34). Conclusions. When comparing the genotyping data of F2 hybrids and the phenotypic manifestation of resistance to leaf rust, it was determined that the microsatellite locus Xwmc344 is closely linked to the Lr13 gene.

Variability of superoxide dismutase isoenzymes in varieties and introgressive lines of wheat differing in drought resistance

Aim. To study the variability of the component composition of superoxide dismutase (SOD) isoenzymes under the influence of hyperthermia and water deficit in introgressive lines of wheat with different drought resistance. Methods. The activity of SOD isoforms was determined using native electrophoresis in an acidic medium in a polyacrylamide gel. Results. Under the impact of stress factors (elevated temperature, water deficit and their combined effects), a redistribution of the relative content of individual SOD isoforms was observed in wheat seedlings of drought-resistant and drought-susceptible genotypes compared to growing under favorable conditions. The most significant decrease occurred due to the joint action of water deficit and increased temperature. Under these conditions, enzyme isoforms of medium electrophoretic mobility practically disappeared. Conclusions. The presence of low-mobility isoforms of SOD in wheat seedlings under the impact of stress factors (elevated temperature and water deficit) can serve as an indicator of drought resistance of wheat plants.

Assessment of the genetic polymorphism of genus Pinus L. the oligotrophic peat bog "Boloto Mshana"

Aim. To evaluate the molecular genetic polymorphism of representatives of the genus Pinus L. on the territory of the "Boloto Mshana" nature reserve, as well as to test the hypothesis about their possible local hybridization Methods. DNA extraction using CTAB and DNeasy Plant Pro Kit, polymerase chain reaction (PCR) with primers for the chloroplast DNA and genes encoding β-tubulin (TBP markers), electrophoresis of DNA in a polyacrylamide gel with silver nitrate staining. Results. DNA profiles of P. mugo, P. sylvestris and probably P. uliginosa (21 samples in total) were analyzed using one chloroplast DNA marker and two DNA markers based on introns of β-tubulin genes (ТВР, сТВР). The intraspecific genetic polymorphism of the species was compared and the probability of the existence of hybrid forms was investigated. Conclusions. The marker based on the 2nd intron of the β-tubulin genes turned out to be the most effective and informative DNA marker for studying representatives of the Pinus L. genus. P. sylvestris showed less genetic polymorphism by the investigated genetic markers, compared to P. mugo and P. uliginosa. Possible hybrid forms (P. mugo ✕ P. sylvestris) or P. uliginosa were also discovered.

Characterization of Bila Tserkva winter common wheat cultivars with respect to marker loci

Aim. We studied special features of the sample of Bila Tserkva winter common wheat cultivars with respect to storage protein loci and some disease resistance genes. Methods. To identify alleles at gliadin and high-molecular-weight glutenin loci, acid polyacrylamide gel electrophoresis and SDS-electrophoresis of grain proteins were carried out. Alleles of markers for the disease resistance genes Lr34, Tsn1, and TDF_076_2D were identified using PCR. Results. The composition and frequencies of alleles at the Gli-1, Glu-1, and Gli-A3 loci were studied in samples of Bila Tserkva cultivars released before 2011 and after 2010. For the latter group of cultivars, alleles of the disease resistance gene markers were identified. We observed a significant increase in the frequency of the allele Gli-A1x(9) and a decrease in the frequency of Gli-A1c, as well as a tendency for an increase in the frequency of Gli-A3c. In the total sample of Bila Tserkva cultivars, nonrandom associations of pairs of certain storage protein alleles were revealed. Conclusions. The group of Bila Tserkva cultivars is similar to the previously studied groups of the Central Forest Steppe of Ukraine with respect to allele set and frequencies of the main prolamin loci, but differs in the frequencies of the disease resistance genes. A special feature of the group of Bila Tserkva cultivars is a high frequency of the allele c at the minor locus Gli-A3, as well as the allele association Gli-A1x Gli-B1l Gli-A3c.

Interaction between modified magnetic nanoparticles and human albumin: Kinetics and isotherm studies and application in protein depletion

Magnetic nanoparticles modified with tetraethyl orthosilicate (Fe3O4@TEOS) and bovine serum albumin (Fe3O4@TEOS@BSA) were evaluated as sorbent in albumin depletion from human serum samples by magnetic dispersive solid phase extraction. Characterization studies were carried out by X-ray diffraction, thermogravimetry, Fourier transform infrared spectroscopy, zeta potential, and scanning electron microscopy. Both nanoparticles also showed high thermal stability and pH-dependent surface charges. The human serum albumin adsorption protocol was optimized using a central composite rotatable design. Nanoparticle mass, pH, and albumin concentration were the most influential variables. Avrami's fractional order and Freundlich isotherm models best fitted the data for human albumin adsorption kinetic and isotherm studies for Fe3O4@TEOS and Fe3O4@TEOS@BSA, and the maximum adsorption capacities were 11.93 and 14.89 mg g−1, respectively. The protein desorption was influenced by the pH of samples and eluent volume. Electrophoresis in a polyacrylamide gel containing sodium dodecyl sulfate showed different patterns of serum protein bands when consecutive depletions were performed. The Fe3O4@TEOS showed greater affinity for HSA and efficiency in depletion. The process was versatile, and the depleted albumin proportion could be controlled by the nanoparticle masses. The proposed method is a powerful sample preparation technique for rapid, reliable, and specific depletion of albumin.