Amplification System Research Articles

Rapid, Simultaneous Detection of Various Biological Toxin Genes Using Multiplex Reverse Transcription Loop-Mediated Isothermal Amplification(RT-LAMP)

Rapid, early, accurate detection and identification of the various pathogenic agents associated with the development of biological weapons is critical in preventing loss of life and limiting the impact of these organisms when used against civilian or military targets. The aim of this study was to produce a system for the simple, rapid, accurate and simultaneous detection and identification of Ricin, Botulinum toxin B and Staphylococcal enterotoxin B as a proof of principle for developing field appropriate reverse transcription loop-mediated isothermal amplification systems for the accurate identification of potential biological threats. These systems were designed to facilitate the identification of potential threats even in remote or resource-limited locations.

An ultrasensitive DNA-enhanced amplification method for detecting cfDNA drug-resistant mutations in non-small cell lung cancer with selective FEN-assisted degradation of dominant somatic fragments.

Blood cell-free DNA (cfDNA) can be a new reliable tool for detecting epidermal growth factor receptor (EGFR) mutations in non-small cell lung cancer (NSCLC) patients. However, the currently reported cfDNA assays have a limited role in detecting drug-resistant mutations due to their deficiencies in sensitivity, stability, or mutation detection rate. We developed an Archaeoglobus fulgidus-derived flap endonuclease (Afu FEN)-based DNA-enhanced amplification system of mutated cfDNA by designing a pair of hairpin probes to anneal with wild-type cfDNA to form two 5'-flaps, allowing for the specific cleavage of wild-type cfDNA by Afu FEN. When the dominant wild-type somatic cfDNA fragments were cleaved by structure-recognition-specific Afu FEN, the proportion of mutated cfDNA in the reaction system was greatly enriched. As the amount of mutated cfDNA in the system was further increased by PCR amplification, the mutation status could be easily detected through first-generation sequencing. In a mixture of synthetic wild-type and T790M EGFR DNA fragments, our new assay still could detect T790M mutation at the fg level with remarkably high sensitivity. We also tested its performance in detecting low variant allele frequency (VAF) mutations in clinical samples from NSCLC patients. The plasma cfDNA samples with low VAF (0.1 and 0.5 %) could be easily detected by DNA-enhanced amplification. This system with enhanced amplification of mutated cfDNA is an effective tool used for the early screening and individualized targeted therapy of NSCLC by providing arapid, sensitive, and economical way for the detection of drug-resistant mutations in tumors.

Detection of foot-and-mouth disease virus RNA using a closed loop-mediated isothermal amplification system.

Foot-and-mouth disease (FMD) is a highly contagious viral disease of cloven-hoofed animals responsible for economic losses that amount to >$20 billion annually. Rapid recognition of FMD cases provides vital information to guide control programmes. A range of point-of-need amplification technologies have been developed which allow sensitive detection of the causative virus (FMDV) in the field at locations remote from laboratories. Here we describe a novel system to detect FMDV RNA using loop-mediated isothermal amplification (LAMP). This test was evaluated using a panel of FMDV isolates (n = 79) and RNA standards demonstrating capability to amplify viral genome directly from clinical material in the absence of nucleic acid extraction. This extraction-free RT-LAMP assay was transferred to a bespoke closed-system lateral flow test (LFT) that was used in combination with a low-cost hand-held heater. Our results show that the RT-LAMP-LFT assay retains a high level of diagnostic and analytical sensitivity when using direct clinical material, with a limit of detection under 80 copies per reaction. Together, our data support the potential for the use of this assay at the point-of-need to facilitate rapid feedback on the status of suspect cases.

An efficient DNAzyme-locked leakless enzyme-free amplification system for alpha-foetoprotein detection in liver cancer and breast cancer.

Alpha-foetoprotein (AFP) is taken as a diagnostic tumor marker for the screening and diagnosis of cancer. Nucleic acid-based isothermal amplification strategies are emerging as a potential technology in early screening and clinical diagnosis of AFP. The leakages between hairpins dramatically increase the background and reduce the sensitivity. Thus, it is necessary to develop some strategies to reduce the leakage for isothermal amplification strategies. A DNAzyme-locked leakless enzyme-free amplification system was developed for AFP detection in liver cancer and breast cancer. AFP could open the apt-hairpin and initiate the catalytic hairpin assembly (CHA) reaction to produce a Y-shaped duplex. Two tails of a Y-shaped duplex cleaved the two kinds of leakless hairpins. Then, the third tail of the Y-shaped duplex catalyzed the second CHA between the cleaved leakless hairpins to recover the fluorescent intensity. The limit of detection reached 5fg/mL by the two levels of signal amplifications. Importantly, the leakless hairpin design effectively reduced leakage between hairpins and weakened the background. In addition, it also showed a great promising potential for AFP detection in early screening and clinical diagnosis.

2.09 µm high-gain, high-energy sub-nanosecond laser amplification system and its application to a millijoule-level mid-infrared ZnGeP2 optical parametric generator.

In this Letter, we report for the first time to our knowledge a 2 mJ-level 2.09 µm Ho:YAG regenerative amplifier (RA) seeded by the first-stage Ho-doped fiber (HDF) preamplifier of a gain-switched laser diode (GSLD). After the single-pass power amplifier (SPPA), the output of a 2.09 µm pulse laser with 1 kHz, 570 ps, and >10 mJ was achieved. The overall gain of the whole amplifier system was greater than 90 dB, providing a novel, stable, and reliable sub-nanosecond (sub-ns) pump source operating at a pulse repetition frequency (PRF) of 1 kHz for an optical parametric generator (OPG) based on ZnGeP2 (ZGP). Specifically, for the ZGP OPG structure, a maximum pulse energy of 1.82 mJ at 3-5 µm had been achieved with an injected pump pulse energy of 5.47 mJ, corresponding to a slope efficiency of 39.5% and an optical-to-optical conversion efficiency (OOCE) of 33.27%.

Kinetics of Strand Displacement Reaction with Acyclic Artificial Nucleic Acids

AbstractToehold‐mediated strand displacement (TMSD) reaction, one of the DNA nanotechnologies, has great potential as s biological programmable platform in the cellular environment. Various artificial nucleic acids have been developed to improve stability and affinity for biological applications. However, the lack of understanding of the kinetics of TMSD reaction among artificial nucleic acids has limited their applications. We herein systematically characterized the kinetics of TMSD reactions with acyclic xeno nucleic acids (XNAs): serinol nucleic acid (SNA), acyclic D‐threoninol nucleic acid (D‐aTNA), and acyclic L‐threoninol nucleic acid (L‐aTNA). We found that the strand displacement reactions by D‐aTNA and by L‐aTNA were highly dependent on temperature. D‐aTNA and L‐aTNA systems were orthogonal to each other, and chirality of the input can be switched by using SNA as an interface. We also applied TMSD reactions of XNAs to a seesaw gate amplification system which utilizes the orthogonality. This work will contribute to the developments of thermoresponsive and bioorthogonal nucleic acid circuits.

Kinetics of Strand Displacement Reaction with Acyclic Artificial Nucleic Acids.

Toehold-mediated strand displacement (TMSD) reaction, one of the DNA nanotechnologies, has great potential as s biological programmable platform in the cellular environment. Various artificial nucleic acids have been developed to improve stability and affinity for biological applications. However, the lack of understanding of the kinetics of TMSD reaction among artificial nucleic acids has limited their applications. We herein systematically characterized the kinetics of TMSD reactions with acyclic xeno nucleic acids (XNAs): serinol nucleic acid (SNA), acyclic D-threoninol nucleic acid (D-aTNA), and acyclic L-threoninol nucleic acid (L-aTNA). We found that the strand displacement reactions by D-aTNA and by L-aTNA were highly dependent on temperature. D-aTNA and L-aTNA systems were orthogonal to each other, and chirality of the input can be switched by using SNA as an interface. We also applied TMSD reactions of XNAs to a seesaw gate amplification system which utilizes the orthogonality. This work will contribute to the developments of thermoresponsive and bioorthogonal nucleic acid circuits.

Minute level ultra-rapid and thousand copies level high-sensitive pathogen nucleic acid identification based on contactless impedance detection microsensor

Early screening for pathogens is crucial during pandemic outbreaks. Nucleic acid testing (NAT) is a valuable method for keeping pathogens from spreading. However, the long detection time and large size of the instruments involved significantly limited the efficiency of detection. This work described an integrated NAT microsensor that facilitated rapid and extremely sensitive detection based on nucleic acid amplification (NAA) on a chip. The biochip consisted of two layers incorporating a heater, a thermometer, an interdigital electrode (IDE) and a reaction chamber. The Pt electrode based heater and thermometer were utilized to maintain a specific temperature for the sample in the chamber. The thermometer exhibited a good linear correlation with a sensitivity of 9.36 Ω/°C and the heater achieved a heating efficiency of approximately 6.5 °C/s. Multiple ions were released during NAA, resulting in a decrease in the impedance of the amplification system solution. A large signal of impedance was generated by the released ions due to its linear correlation with the logarithm of the ion concentration. With this detection principle, IDE was employed for real-time monitoring of the in-chip reaction system impedance and NAA process. Specific nucleic acids from two pathogens (SARS-CoV-2, Vibrio vulnificus) were detected with this microsensor. The samples were qualitatively analyzed on microchip within 3 min, with a limit of detection (LOD) of 103 copies/μL. The proposed sensor presented several advantages, including reduced NAT time and increased sensitivity. Consequently, it has shown significant potential in rapid and high-quality nucleic acid testing for the field of epidemic prevention.

High-sensitivity whole-mount in situ Hybridization of Mouse Oocytes and Embryos Visualizes the Super-resolution Structures and Distributions of mRNA Molecules

Mammalian oocytes accumulate more than ten thousand mRNAs, of which three to four thousand mRNAs are translationally repressed. The timings and sites of translational activation of these dormant mRNAs are crucial for promoting oocyte maturation and embryonic development. How these mRNAs are accumulated and distributed in oocytes is therefore a fundamental issue to be explored. A method that enables visualization of mRNA molecules with high resolution in a simple manner would be valuable for understanding how oocytes accumulate and regulate the dormant mRNAs. We have developed a highly sensitive whole-mount in situ hybridization method using in vitro-synthesized RNA probes and the tyramide signal amplification (TSA) system optimized for mouse oocytes and embryos. By using this method, Pou5f1/Oct4, Emi2, and cyclin B1 mRNAs were detected in immature oocytes and 2-cell stage embryos. Confocal microscopy showed that these mRNAs formed granular structures in the oocyte cytoplasm. The structures of Pou5f1/Oct4 and cyclin B1 mRNAs persisted in 2-cell stage embryos. Pou5f1/Oct4 RNA granules exhibited a solid-like property in immature oocytes and became liquid-like droplets in 2-cell stage embryos. Double-staining of cyclin B1 mRNA with Emi2 or Pou5f1/Oct4 mRNA revealed that these mRNAs were distributed as different RNA granules without overlapping each other and that the size of cyclin B1 RNA granules tended to be larger than that of Emi2 RNA granules. The structures and distribution patterns of these mRNAs were further analyzed by N-SIM super-resolution microscopy. This analysis revealed that the large-sized RNA granules consist of many small-sized granules, suggesting the accumulation and regulation of dormant mRNAs as basal-sized RNA granules. The method established in this study can easily visualize the structure and distribution of mRNAs accumulated in mammalian oocytes and embryos with high sensitivity and super-resolution. This method is useful for investigating the cellular and molecular mechanisms of translational control of mRNAs by which maturation and early developmental processes are promoted.

Allosteric probe-initiated double rolling circle amplification in tandem for sensitive whole-cell detection of E. coli O157:H7

The development of facile, reliable nucleic acid amplification assays for pathogenic bacteria is paramount but still poses a challenge. Here we present a molecular amplification system that integrates the whole-cell recognition of allosteric probes with cascaded rolling circle amplification. Allosteric probe-initiated double rolling circle amplification (AID-RCA) assay involves a three-step process that utilizes allosteric probes, dumbbell templates, and circular templates to recognize, amplify, and detect target bacteria, E. coli O157:H7. The allosteric probes are designed to recognize E. coli O157:H7 without bacterial isolation and nucleic acid extraction. Subsequently, the two-stage in-tandem amplification, that is, dumbbell template-based exponential-rolling circle amplification (DT-RCA) and circular template-based linear-rolling circle amplification (CT-RCA), is initiated and yields powerful fluorescence signals using molecular beacon as a signal probe. The symmetric structure of the dumbbell template endows the AID-RCA assay with good specificity. Cascaded signal amplification boosts its sensitivity. The AID-RCA assay demonstrated high sensitivity in the quantitative detection of E. coli O157:H7(limit of detection 1.6 CFU·mL−1). Compared to real-time PCR, the AID-RCA system exhibited higher sensitivity for low concentrations of E. coli O157:H7. Furthermore, the AID-RCA system can selectively differentiate E. coli O157:H7 from other pathogenic bacteria in complex samples such as drinking water and milk. In a blind validation study using milk samples, the AID-RCA system effectively distinguished between contaminated and pasteurized milk samples. Our method is expected to hold great potential for developing precise, reliable, and efficient analytical platforms to combat pathogenic bacterial pollution and safeguard human health.

A Dual-Recognition Fluorescence Enzyme-Linked Immunosorbent Assay for Specific Detection of Intact Lipid Nanoparticles via a Localized Scaffolding Autocatalytic DNA Circuit Amplifier.

Lipid nanoparticles (LNPs) are emerging as one of the most promising drug delivery systems. The long-circulating effect of intact LNPs (i-LNPs) is the key to efficacy and toxicity in vivo. However, the significant challenge is specific and sensitive detection of i-LNPs. Herein, a dual-recognition fluorescence enzyme-linked immunosorbent assay (DR-FELISA) was developed to directly isolate and detect i-LNPs by combining dual-recognition separation with a one-step signal amplification strategy. The microplates captured and enriched i-LNPs through antibody-antigen reaction. Dual-chol probes were spontaneously introduced into the lipid bilayer of captured i-LNPs, converting the detection of i-LNPs into the detection of double-cholesterol probes. Finally, the end of the dual-chol probes initiated the localized scaffolding autocatalytic DNA circuits (SADC) system for further signal amplification. The SADC system provides a sensitive and efficient amplifier through localized network structures and self-assembled triggers. Simultaneous recognition of i-LNPs surface PEG-lipid and lipid bilayer structures significantly eliminates interference from biological samples. i-LNPs were detected with high selectivity, ranging from 0.2 to 1.25 mg/mL with a limit of detection of 0.1 mg/mL. Moreover, this method allows the isolation and quantitative analysis of different formulations of i-LNPs in serum samples with a satisfactory recovery rate ranging from 94.8 to 116.3%. Thus, the DR-FELISA method provides an advanced platform for the exclusive and sensitive detection of i-LNPs, providing new insights for the study of the quality and intracorporal process of complex formulations.

Разработка датчика для анализа процессов взаимодействия заряженных частиц и контроля содержания радиоактивных элементов

Of great interest are the processes associated with the results of interaction of charged particles accelerated to energies of several giga-electron volts; they can be applied in the most modern science and technology. To obtain maximum information about the interaction of charged particles in accelerators, special radiation sensors are required that operate in the corresponding sections of the spectrum, both simple ones, such as cathode and photo luminescent detectors, and complex highly sensitive vacuum equipment that not only records even very weak radiation, but also significantly amplifies it. In this work, an automated calculation of the electron-optical and amplifying system of the device will be carried out, a photocathode will be selected, a design of the vacuum block of the sensor will be proposed, and its electric power supply modes will be indicated. Materials and methods. Theoretical analysis based on the studied specialized literature and practical work experience. Development of high-tech products using the automated design system. Optimization and system analysis. Results. An automated calculation of the electron-optical and amplifying system of a photomultiplier-type device with micro-channel amplification was performed, a photocathode was selected, and conditions for minimizing noise during radiation monitoring were provided. Based on the calculations performed, a design of the vacuum block of the sensor was proposed, its electrical power supply modes were specified, and technological issues were considered. Discussion. The modes of operation of the radiation sensor were identified, providing conditions for minimizing noise in operation, concerning the energy of photoelectrons approaching the secondary electron multipliers, the configuration of the signal beam. The design of the radiation sensor of the PMT type was developed, providing the required high levels of the main parameters: sensitivity threshold, signal gain coefficient, photoelectron collection coefficient, anode dark current, and response speed. Conclusion. A critical analysis of approaches to the study of the interaction process of charged particles and experimental means was carried out. The design of a highly efficient radiation sensor was developed, its optimization was carried out in terms of increasing the efficiency of interaction of photoelectrons with the amplification system. An assembly drawing of the vacuum block design was developed, the most important technological issues of its manufacture were resolved. Resume. The results of the work can be useful in setting up experiments to study the processes of interaction of particles accelerated to energies of several giga-electron volts, as well as in the development of devices using ultraviolet radiation, including medical ones.

Nucleic Acid Target Sensing Using a Vibrating Sharp-Tip Capillary and Digital Droplet Loop-Mediated Isothermal Amplification (ddLAMP).

Nucleic acid tests are key tools for the detection and diagnosis of many diseases. In many cases, the amplification of the nucleic acids is required to reach a detectable level. To make nucleic acid amplification tests more accessible to a point-of-care (POC) setting, isothermal amplification can be performed with a simple heating source. Although these tests are being performed in bulk reactions, the quantification is not as accurate as it would be with digital amplification. Here, we introduce the use of the vibrating sharp-tip capillary for a simple and portable system for tunable on-demand droplet generation. Because of the large range of droplet sizes possible and the tunability of the vibrating sharp-tip capillary, a high dynamic range (~2 to 6000 copies/µL) digital droplet loop-mediated isothermal amplification (ddLAMP) system has been developed. It was also noted that by changing the type of capillary on the vibrating sharp-tip capillary, the same mechanism can be used for simple and portable DNA fragmentation. With the incorporation of these elements, the present work paves the way for achieving digital nucleic acid tests in a POC setting with limited resources.

Genetic landscape of the people of Bangladesh depicted with 17 Y-Chromosome-Specific microsatellites

Seventeen microsatellite loci from the non-recombining region of the human Y chromosome were typed using AmpFlSTR® Yfiler® PCR amplification systems in 404 male subjects belonging to the three largest ethnic groups in Bangladesh. A total of 150 haplotypes from the Chakma, 144 from the Tripura, and 110 from the Khasia were detected with a corresponding discrimination capacity of 73.885%, 65.563%, and 81.250%, respectively. The highest allele frequency of 0.828 was detected in DYS391 locus in the Tripuras, while the lowest allele frequency of 0.009 was detected at the same locus for the Chakma population. The highest gene diversity (0.964) was observed at DYS385a/b locus in the Khasias, while the lowest gene diversity (0.301) was detected at DYS391 locus in Tripuras population. The overall haplotype diversity for the studied populations was 0.986141. Both the Neighbour-Joining tree and pairwise genetic distances showed that Chakma lies closer to a clade consisting of Tripuras (Khagrachari, Bangladesh) and Tripuri (Tripura, India). In contrast, the Khasias demonstrated a close affinity with the Oraon (Chhattisgarh, India), followed by the Santals. The Y-STR haplotype matching probabilities within and between populations demonstrated that the Chakma, Tripura, and Khasia were 100% genetically distinct. The studied ethnic populations exhibited higher frequency for haplogroups L and Q as opposed to haplogroups R1a, H, and L found in the mainstream Bengali population. The Median-joining networking showed haplogroups L and R1a have the most compact clustering within populations, followed by haplogroups Q and H. The presence of haplogroup R1a suggests that Bengali may have originated through west-to-east migration, whereas haplogroups L and Q distribution in the studied tribes reveal a very significant affinity with the South-East Asian populations and may have shared a common ancestral origin with the Mongoloid stock populations. Bioresearch Commu. 10(2): 1482-1488, 2024 (July)

Linear dispersion (GDD) design using grating group

Precise control of dispersion output holds paramount significance across domains such as optical fiber communication, time stretching, and spectral interferometric ranging. In comparison to other dispersion elements, like prisms, gratings are widely applied in the field of dispersion control due to their advantages of broad spectral range, tunability, and high resolution. Moreover, linear dispersion is the most desired characteristic by designers in most cases. Here, we develop a dispersion model for grating groups to determine the optimal structural parameters for achieving linear dispersion in high-order grating arrays. Based on our model, we provide corresponding parameter selection methods that allow for quantitative design of the size and slope of output dispersion by adjusting input parameters such as angle, distance, and parallelism. Additionally, we experimentally establish a dispersion interferometry structure based on the grating ensemble that validates our proposed approach's capability for linear dispersion output (linearity better than 0.9998). We believe that our approach is universally significant and contributes to enhancing the performance of dispersion interferometric measurement systems, chirp amplification systems, and other related systems.

High-Power GHz Burst-Mode All-Fiber Laser System with Sub 300 fs Pulse Duration

An all-fiber low-repetition-rate SESAM mode-locked fiber oscillator combined with a dispersion-managed active fiber loop produces a flexible GHz burst-mode laser source. The high-power output is then produced by amplifying the GHz burst-mode laser source using an all-fiber chirped-pulse amplification system. Then, the laser is compressed using a grating pair compressor; a maximum amplified power of 97 W is obtained. This results in a compressed high power of 82.07 W with a power stability RMS of 0.09% and beam quality better than 1.2. Accurate dispersion control allows for the production of a high-quality pulse duration of 265 fs.

PACRAT: pathogen detection with aptamer-observed cascaded recombinase polymerase amplification-in vitro transcription.

The SARS-CoV-2 pandemic underscored the need for early, rapid, and widespread pathogen detection tests that are readily accessible. Many existing rapid isothermal detection methods use the recombinase polymerase amplification (RPA), which exhibits polymerase chain reaction (PCR)-like sensitivity, specificity, and even higher speed. However, coupling RPA to other enzymatic reactions has proven difficult. For the first time, we demonstrate that with tuning of buffer conditions and optimization of reagent concentrations, RPA can be cascaded into an in vitro transcription reaction, enabling detection using fluorescent aptamers in a one-pot reaction. We show that this reaction, which we term PACRAT (pathogen detection with aptamer-observed cascaded recombinase polymerase amplification-in vitro transcription) can be used to detect SARS-CoV-2 RNA with single-copy detection limits, Escherichia coli with single-cell detection limits, and 10-min detection times. Further demonstrating the utility of our one-pot, cascaded amplification system, we show PACRAT can be used for multiplexed detection of the pathogens SARS-CoV-2 and E. coli, along with multiplexed detection of two variants of SARS-CoV-2.

In Situ Complexation of sgRNA and Cas12a Improves the Performance of a One-Pot RPA-CRISPR-Cas12 Assay.

Due to their ability to selectively target pathogen-specific nucleic acids, CRISPR-Cas systems are increasingly being employed as diagnostic tools. "One-pot" assays that combine nucleic acid amplification and CRISPR-Cas systems (NAAT-CRISPR-Cas) in a single step have emerged as one of the most popular CRISPR-Cas biosensing formats. However, operational simplicity comes at a cost, with one-pot assays typically being less sensitive than corresponding two-step NAAT-CRISPR-Cas assays and often failing to detect targets at low concentrations. It is thought that these performance reductions result from the competition between the two enzymatic processes driving the assay, namely, Cas-mediated cis-cleavage and polymerase-mediated amplification of the target DNA. Herein, we describe a novel one-pot RPA-Cas12a assay that circumvents this issue by leveraging in situ complexation of the target-specific sgRNA and Cas12a to purposefully limit the concentration of active Cas12a during the early stages of the assay. Using a clinically relevant assay against a DNA target for HPV-16, we show how this in situ format reduces competition between target cleavage and amplification and engenders significant improvements in detection limit when compared to the traditional one-pot assay format, even in patient-derived samples. Finally, to gain further insight into the assay, we use experimental data to formulate a mechanistic model describing the competition between the Cas enzyme and nucleic acid amplification. These findings suggest that purposefully limiting cis-cleavage rates of Cas proteins is a viable strategy for improving the performance of one-pot NAAT-CRISPR-Cas assays.

Platinum Nanozyme Probes for Cellular Imaging by Electron Microscopy

The highly efficient peroxidase‐like activity of platinum nanozymes (3–20 nm size) is exploited within the complex cellular environment to catalyze the oxidation of the DAB substrate, producing an electron‐dense signal around the nanozyme surface, upon osmium staining. It is proved that such nanozyme amplification can achieve a catalytic signal enhancement up to 10‐fold, enabling the quick detection of the Pt particles (even of 3 nm size) by transmission electron microscopy (TEM) also at low magnification and across wide fields of view in the intricate intracellular milieu. The developed procedure is ideally suited to overcome standard amplification strategies currently used in TEM analysis, such as gold or silver enhancements. Furthermore, the wide versatility of the Pt‐nanozyme probes in TEM imaging is demonstrated in immuno‐EM and protein trafficking studies, showing their potential to track the subcellular localization of target biomolecules at both low and high magnifications. These results suggest that the use of nanozymes might represent a paradigm shift in the conventional amplification systems currently employed in electron microscopy for cellular analyses, offering enhanced imaging capabilities.

Prospects for the development of drivetrain systems in trucks

The article presents the directions of development of drive systems in trucks. A database of vehicles and their technical data was developed. This data contains information about the drivetrain configuration. The external characteristics of the engines and the structure of the drivetrain were analyzed. It was found that the factors determining development are primarily economic and ecological aspects. It was noticed that, based on the configuration of the drive system, vehicles can be divided according to their purpose and type of transport. Vehicles intended for international transport are characterized by low gears and engines that achieve high torque at low engine speeds. Construction vehicles and vehicles intended for oversize transport are characterized by large gear ratios and torque amplification systems. The structure of electric vehicle drive systems is different from conventional vehicles. Gearboxes usually have two gears, drive axles may have integrated engines located next to the main gear or in the final drive position. The study also made it possible to outline the problem of the short range of such cars and the need for further work on increasing it.