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Immunoassay Systems for Detection and Quantitative Determination of Cefalexin
IF 1 4区 生物学
Applied Biochemistry and Microbiology Pub Date : 2024-12-18 DOI: 10.1134/S0003683824605638
T. S. Serchenya, I. V. Harbachova, I. I. Vashkevich, O. V. Sviridov
{"title":"Immunoassay Systems for Detection and Quantitative Determination of Cefalexin","authors":"T. S. Serchenya,&nbsp;I. V. Harbachova,&nbsp;I. I. Vashkevich,&nbsp;O. V. Sviridov","doi":"10.1134/S0003683824605638","DOIUrl":"10.1134/S0003683824605638","url":null,"abstract":"<p>Beta-lactam antibiotic cephalexin is poorly recognized by bacterial beta-lactam binding proteins and therefore cannot be reliably detected by a group-specific receptor assay. For effective and rapid monitoring of cephalexin concentration in food products, the test systems based on high-affinity and selective binding of this antibiotic to polyclonal antibodies have been developed. The principle of the method for the test systems of enzyme-linked immunosorbent assay (ELISA) and lateral flow immunoassay (LFIA) is based on the competitive interaction of (a) cephalexin, potentially contained in the samples, and (b) a cephalexin-protein conjugate immobilized in the wells of a microplate or on the membrane of a test strip, with (c) polyclonal antibodies against cephalexin conjugated with peroxidase or adsorbed on gold nanoparticles. The high specificity of the obtained polyclonal antibodies against cephalexin and the absence of cross-reactivity to a number of cephalosporins, penicillins, and antibiotics of other classes have been confirmed. In the ELISA system in a buffer solution and in a milk matrix, the detection limit, the IC<sub>50</sub> value, and the range of detected concentrations were 0.02, 1.0 and 0.025–100 ng/mL respectively. The limit of detection for visual and instrumental determination of cephalexin in the LFIA system were 1.5 and 0.1 ng/mL, and the working range of quantitatively measured concentrations was from 0.05 to 1.5 ng/mL. For both systems, the coefficient of variation of measurements was in the range of 2.5–8.0%. The test systems allow to detect cephalexin in milk without sample preparation. The recovery of cephalexin added to the milk samples was 90.0–106.7%. The presented developments can serve as the basis for kits of reagents for monitoring the content of cephalexin in food products.</p>","PeriodicalId":466,"journal":{"name":"Applied Biochemistry and Microbiology","volume":"60 6","pages":"1416 - 1427"},"PeriodicalIF":1.0,"publicationDate":"2024-12-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142844787","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Adaptation of Erythrocytes: The Role of Hemoglobin, Nitric Oxide, and Methylglyoxal
IF 1 4区 生物学
Applied Biochemistry and Microbiology Pub Date : 2024-12-18 DOI: 10.1134/S0003683824605468
O. V. Kosmachevskaya, A. F. Topunov
{"title":"Adaptation of Erythrocytes: The Role of Hemoglobin, Nitric Oxide, and Methylglyoxal","authors":"O. V. Kosmachevskaya,&nbsp;A. F. Topunov","doi":"10.1134/S0003683824605468","DOIUrl":"10.1134/S0003683824605468","url":null,"abstract":"<p>All living systems are characterized by fundamental properties such as the ability to adaptation and self-regulation. Mammalian nonnuclear erythrocytes also have the ability to adapt to external effects, but their regulatory capabilities are limited by cytoplasmic mechanisms, including phase transitions of proteins and membranes. This is one of the most ancient mechanisms of adaptation of living systems to external and internal conditions. Erythrocytes under changes in plasma composition, aging, and energy depletion, undergo a reversible morpho-functional transformation, the transition from a discocyte to an echinocyte. The metabolic shifts occurring in this case correspond to a complex of universal changes that take place during erythrocyte transition to metabolic depression. As a rule, echinocytosis is considered as a pathological process preceding eryptosis and hemolysis. However, it can be also considered as the first stage of the implementation of a universal program of passive cell adaptation, the ultimate goal of which is to transfer the system to a state of suspended animation. The energy status of an erythrocyte is determined by the equilibrium of soluble and membrane-bound hemoglobin (Hb) forms. Compounds with pronounced electrophilic properties—nitric oxide and methylglyoxal—affecting this equilibrium can induce cell’s transition from one metabolic state to another. The mechanism of their action is largely related to the modification of thiol groups of membrane and cytoskeleton proteins, including reactive SH-groups of Hb. It seems relevant to consider their effect on the state of Hb and erythrocytes.</p>","PeriodicalId":466,"journal":{"name":"Applied Biochemistry and Microbiology","volume":"60 6","pages":"977 - 992"},"PeriodicalIF":1.0,"publicationDate":"2024-12-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142845040","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Practical Achievements of Laboratory Evolution
IF 1 4区 生物学
Applied Biochemistry and Microbiology Pub Date : 2024-12-18 DOI: 10.1134/S000368382460533X
Ya. E. Dunaevsky, O. A. Kudryavtseva, S. M. Agroskin, A. A. Gasparyan, M. A. Belozersky
{"title":"Practical Achievements of Laboratory Evolution","authors":"Ya. E. Dunaevsky,&nbsp;O. A. Kudryavtseva,&nbsp;S. M. Agroskin,&nbsp;A. A. Gasparyan,&nbsp;M. A. Belozersky","doi":"10.1134/S000368382460533X","DOIUrl":"10.1134/S000368382460533X","url":null,"abstract":"<p>Adaptive Laboratory Evolution (ALE) is a novel approach to generating microbial strains with desired characteristics and producing value-added products. Additionally, ALE is employed as a means of enhancing comprehension of the genetic and/or metabolic pathways of evolution. The objective of this review is to analyze the results of studies that elucidate and demonstrate the potential of microorganisms as model objects for laboratory evolutionary experiments. These experiments are becoming increasingly prevalent in the study of adaptation, the evaluation of evolutionary dynamics, and the testing of various evolutionary hypotheses. Concurrently, ALE has demonstrated itself to be a promising and efficacious methodology, which, when employed for biotechnological applications, has already resulted in the generation of novel and useful microbial strains. It is important to note that the current achievements not only demonstrate the power and versatility of this approach but also highlight a number of unanswered questions. The conclusions drawn on the basis of ALE require a cautious interpretation of the results obtained.</p>","PeriodicalId":466,"journal":{"name":"Applied Biochemistry and Microbiology","volume":"60 6","pages":"1010 - 1021"},"PeriodicalIF":1.0,"publicationDate":"2024-12-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142845041","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Mechanisms of the Antimicrobial Action of Fatty Acids: A Review
IF 1 4区 生物学
Applied Biochemistry and Microbiology Pub Date : 2024-12-18 DOI: 10.1134/S0003683824605158
E. S. Obukhova, S. A. Murzina
{"title":"Mechanisms of the Antimicrobial Action of Fatty Acids: A Review","authors":"E. S. Obukhova,&nbsp;S. A. Murzina","doi":"10.1134/S0003683824605158","DOIUrl":"10.1134/S0003683824605158","url":null,"abstract":"<p>Among the diverse biological activities of fatty acids (<b>FAs</b>), the ability to kill or inhibit the growth of microorganisms can be distinguished. Even though the antibacterial mechanisms of fatty acids are not fully understood, the most common target of action is the cell membrane, where FAs mediate an increase in permeability and subsequent cell lysis, leading to disruption of the electron transport chain, uncoupling of oxidative phosphorylation, and inhibition of enzymatic activity and nutrient intake. In addition to acting on cell membranes, FAs can disrupt the metabolic processes of microorganisms, inhibit DNA/RNA replication, and affect the expression of virulence genes. In addition, nontraditional mechanisms of the antimicrobial action of FAs are currently being described, such as the inhibition of horizontal gene transfer, quorum sensing, and the disruption of the efflux pump. The variety of antimicrobial mechanisms and the wide range of their activity determine the high biotechnological potential of fatty acids and make further studies of the mechanisms of action on biological systems relevant.</p>","PeriodicalId":466,"journal":{"name":"Applied Biochemistry and Microbiology","volume":"60 6","pages":"1035 - 1043"},"PeriodicalIF":1.0,"publicationDate":"2024-12-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142845042","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Development of Methods for Protecting Mammoth Tusks from Biocontamination 开发保护猛犸象头骨免受生物污染的方法
IF 1 4区 生物学
Applied Biochemistry and Microbiology Pub Date : 2024-12-18 DOI: 10.1134/S000368382460461X
E. S. Petukhova, L. A. Erofeevskaya, T. A. Isakova, V. V. Pavlova, T. M. Solovev, G. V. Botvin, A. A. Chirikov, V. V. Petrov
{"title":"Development of Methods for Protecting Mammoth Tusks from Biocontamination","authors":"E. S. Petukhova,&nbsp;L. A. Erofeevskaya,&nbsp;T. A. Isakova,&nbsp;V. V. Pavlova,&nbsp;T. M. Solovev,&nbsp;G. V. Botvin,&nbsp;A. A. Chirikov,&nbsp;V. V. Petrov","doi":"10.1134/S000368382460461X","DOIUrl":"10.1134/S000368382460461X","url":null,"abstract":"<p>The results of studies of the effectiveness of various methods of protecting mammoth tusks from microbiological contamination under long-term storage conditions are presented. When analyzing swabs taken from the surfaces of mammoth tusk samples before exposure, the landscape of isolated microorganisms was dominated by bacteria of the genera <i>Bacillus</i>, <i>Chryseobacterium</i>, and <i>Enterobacter</i> with biocontamination of bone tissue caused mainly by microorganisms from the external environment (air, anthropogenic impact, etc.). Of the proposed options for protection against biocontamination, the method that showed the greatest effectiveness was the treatment of mammoth tusks with a hydrophobic-biocide composition, followed by mechanical isolation in plastic film. When using this method of protection, the total contamination of mammoth tusk samples after six months of exposure in various conditions decreased by 4–5 times.</p>","PeriodicalId":466,"journal":{"name":"Applied Biochemistry and Microbiology","volume":"60 6","pages":"1275 - 1281"},"PeriodicalIF":1.0,"publicationDate":"2024-12-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142844950","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
An Escherichia coli Strain for Plasmid DNA Production with a Low Endotoxin Level 用于生产内毒素水平较低的 DNA 质粒的大肠杆菌菌株
IF 1 4区 生物学
Applied Biochemistry and Microbiology Pub Date : 2024-12-18 DOI: 10.1134/S0003683824604517
D. D. Kharlampieva, P. A. Bobrovsky, E. N. Grafskaia, V. N. Lazarev, V. A. Manuvera
{"title":"An Escherichia coli Strain for Plasmid DNA Production with a Low Endotoxin Level","authors":"D. D. Kharlampieva,&nbsp;P. A. Bobrovsky,&nbsp;E. N. Grafskaia,&nbsp;V. N. Lazarev,&nbsp;V. A. Manuvera","doi":"10.1134/S0003683824604517","DOIUrl":"10.1134/S0003683824604517","url":null,"abstract":"<p>Plasmid DNA (pDNA) is one of the possible types of vectors used in gene therapy. <i>Escherichia coli</i> is the most common bacterium for the production of pDNA. Lipopolysaccharides are the main component of the outer membrane of <i>E. coli</i>. Lipopolysaccharides are strong endotoxins, and their presence in pharmacological substances is highly undesirable. One of the approaches to eliminating endotoxin contamination of target substances is knocking out the genes of the lipopolysaccharide biosynthesis pathway. Using the CRISPR/Cas9 genome editing technology, we obtained a mutant form of the <i>E. coli</i> strain SCS110. We knocked out the genes <i>kdsD, gutQ, lpxM, lpxL, lpxP, pagP,</i> and <i>eptA</i>. As a result, lipopolysaccharide contamination of pDNA samples isolated by different methods from the mutant <i>E. coli</i> SCS 110 was significantly lower (19 to 295 times) than in pDNA samples from the original strain.</p>","PeriodicalId":466,"journal":{"name":"Applied Biochemistry and Microbiology","volume":"60 6","pages":"1147 - 1152"},"PeriodicalIF":1.0,"publicationDate":"2024-12-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1134/S0003683824604517.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142844951","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Luminescent Nanozyme-Based Sensors for Bacteria Detection 基于发光纳米酶的细菌检测传感器
IF 1 4区 生物学
Applied Biochemistry and Microbiology Pub Date : 2024-12-18 DOI: 10.1134/S0003683824606139
D. S. Chumakov, S. S. Evstigneeva, O. I. Guliy
{"title":"Luminescent Nanozyme-Based Sensors for Bacteria Detection","authors":"D. S. Chumakov,&nbsp;S. S. Evstigneeva,&nbsp;O. I. Guliy","doi":"10.1134/S0003683824606139","DOIUrl":"10.1134/S0003683824606139","url":null,"abstract":"<p>Bacterial infections still remain one of the main problems for healthcare, due to difficulties in timely diagnosis and increasing microbial resistance. Based on advanced nanotechnologies, in particular, nanozymes, as artificial enzymes with a wide spectrum of action, it is possible to create completely new tools for detecting infectious agents. Recent advances in the development of luminescent nanozymes capable of identifying pathogenic bacteria are discussed. The basic information about nanozymes with luminescent properties and their advantages over other variants of nanozymes are considered, and the key mechanisms for generating a luminescent signal that make it possible to detect bacterial cells are revealed. The main focus of the review is on the comprehensive characterization of luminescent sensor systems developed to date, which are aimed at visualizing bacteria, both in a free-living form and in biofilm formation. In addition, a description is given of nanozyme-based sensors that selectively detect biomarkers of bacterial infections using a luminescent signal. This review is focused on further progress in the development of luminescent nanozymes for the detection of bacterial pathogens, as well as on the search for innovative nanozyme technologies that expand the capabilities of clinical microbiology.</p>","PeriodicalId":466,"journal":{"name":"Applied Biochemistry and Microbiology","volume":"60 6","pages":"1022 - 1034"},"PeriodicalIF":1.0,"publicationDate":"2024-12-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142845043","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Optimization of Aerobic Synthesis of Fumaric Acid from Glucose by a Recombinant Escherichia coli Strain Functioning in a Whole-Cell Biocatalyst Mode
IF 1 4区 生物学
Applied Biochemistry and Microbiology Pub Date : 2024-12-18 DOI: 10.1134/S0003683824605729
A. Yu. Skorokhodova, A. A. Stasenko, A. Yu. Gulevich
{"title":"Optimization of Aerobic Synthesis of Fumaric Acid from Glucose by a Recombinant Escherichia coli Strain Functioning in a Whole-Cell Biocatalyst Mode","authors":"A. Yu. Skorokhodova,&nbsp;A. A. Stasenko,&nbsp;A. Yu. Gulevich","doi":"10.1134/S0003683824605729","DOIUrl":"10.1134/S0003683824605729","url":null,"abstract":"<p>Biocatalytic synthesis of fumaric acid from glucose by the previously engineered <i>E. coli</i> strain FUM1.0 (pMW119-<i>kgd</i>) (<i>E. coli</i> MG1655 ∆<i>ackA-pta</i>, ∆<i>ldhA</i>, ∆<i>adhE</i>, ∆<i>ptsG</i>, P<sub>L</sub>-<i>glk</i>, P<sub><i>tac</i></sub>-<i>galP</i>, ∆<i>fumB</i>, ∆<i>fumAC</i>, <i>poxB::</i>P<sub>L</sub>-<i>pycA</i><sup><i>Bs</i></sup>, pMW119-<i>kgd</i>) was optimized. The maximal yield of the target substance was achieved upon its synthesis through a variant of the tricarboxylic acid cycle mediated by the action of heterologous 2-ketoglutarate decarboxylase. The enhanced expression of the genes encoding components of the succinate dehydrogenase enzymatic complex did not markedly affect the biosynthetic characteristics of the producing strain. A positive effect of decreasing the intracellular ATP supply on the conversion of the carbohydrate substrate into the target product was demonstrated. The activation of the futile cycle of pyruvate–phosphoenolpyruvate–pyruvate due to an increase in the expression of the <i>ppsA</i> gene led to a slight increase in the yield of fumaric acid. Upon uncoupling the H<sup>+</sup>-ATP synthase complex subunits resulting in ATP formation cessation via oxidative phosphorylation, due to deletion of the <i>atpFH</i> genes, the molar yield of fumaric acid from glucose demonstrated by the strain functioning in the whole-cell biocatalyst mode reached 92%.</p>","PeriodicalId":466,"journal":{"name":"Applied Biochemistry and Microbiology","volume":"60 6","pages":"1096 - 1103"},"PeriodicalIF":1.0,"publicationDate":"2024-12-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1134/S0003683824605729.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142845039","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Structural Diversity of Fungal Elicitors
IF 1 4区 生物学
Applied Biochemistry and Microbiology Pub Date : 2024-12-18 DOI: 10.1134/S0003683824605973
G. M. Frolova, A. O. Berestetskiy
{"title":"Structural Diversity of Fungal Elicitors","authors":"G. M. Frolova,&nbsp;A. O. Berestetskiy","doi":"10.1134/S0003683824605973","DOIUrl":"10.1134/S0003683824605973","url":null,"abstract":"<p>This article discusses recent advances in the study of fungal biogenic elicitors, examines their structural diversity and mechanisms of immune response, and analyzes the prospects for commercial application. Attention is focused on hydrophilic elicitors (exo- and endo-β-glycans, secreted peptides), lipophilic elicitors (ergosterol and cerebrosides), and secondary metabolites (phytotoxins), which are involved in the intercellular interactions of the pathogen and the host plant and have a great immunostimulating potential. The use of nanoparticles and nanoemulsions that enhance the bioavailability of elicitors as a promising strategy in the preparation of their formulations is discussed.</p>","PeriodicalId":466,"journal":{"name":"Applied Biochemistry and Microbiology","volume":"60 6","pages":"1061 - 1073"},"PeriodicalIF":1.0,"publicationDate":"2024-12-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142845044","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Passive Oral Immunization as a Promising Tool for Treating Rotavirus Infections in Newborns
IF 1 4区 生物学
Applied Biochemistry and Microbiology Pub Date : 2024-12-18 DOI: 10.1134/S0003683824605079
D. S. Shlykova, M. S. Yurkova, A. V. Lipkin, A. N. Fedorov
{"title":"Passive Oral Immunization as a Promising Tool for Treating Rotavirus Infections in Newborns","authors":"D. S. Shlykova,&nbsp;M. S. Yurkova,&nbsp;A. V. Lipkin,&nbsp;A. N. Fedorov","doi":"10.1134/S0003683824605079","DOIUrl":"10.1134/S0003683824605079","url":null,"abstract":"<p>The development of antiviral drugs due to the high risk of rotavirus infections, especially for newborns, has attracted a lot of attention. This review is devoted to consideration of additional and promising tools for controlling human rotavirus infection (HRI) in newborns—passive oral immunization. Vaccination of animals allows us to obtain immunoglobulins in the form of colostrum and use them against human diseases. The use of bovine colostrum as an additional therapeutic agent against HRI in newborns is analyzed.</p>","PeriodicalId":466,"journal":{"name":"Applied Biochemistry and Microbiology","volume":"60 6","pages":"1087 - 1095"},"PeriodicalIF":1.0,"publicationDate":"2024-12-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142845047","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
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