Electrophoresis Technique Research Articles

Surface Functionalization of Gold Nanoparticles Using Alkyne Derivatives: Applications in Chemical Sensing.

Colloidal gold nanoparticles (AuNPs) are important nanomaterials for chemical sensing and therapeutics. For their application, it is vital to develop a reliable and robust surface functionalization method that can be applied to diverse functional molecules and offer better stability under harsh biological conditions. Currently, thiol (SH) is the most commonly used functional group for forming stable covalent bonds with AuNPs. However, thiolated molecules typically require complicated preparation procedures, are susceptible to oxidation, and are not compatible with many electrophiles and reducing groups. In this study, we report that surface functionalization of AuNPs can be achieved using alkyne derivatives, which exhibit several advantages over classical thiolation and peptide-bond methods, including straightforward preparation of alkyne derivatives, rapid and simple conjugation in buffers and complex media, higher conjugation efficiency, long-term stability, and resistance to decomposition under harsh conditions. Several alkynylated biotin and fluorescein derivatives are prepared, and the alkynylated-AuNPs are characterized using a lateral flow assay, gel electrophoresis, and spectroscopy techniques to investigate the conjugation efficiencies, size distributions, protein interaction properties, and binding mode of the Au-alkyne bond. We also demonstrate that alkynylated-AuNPs can be used for the sensitive detection of hydrogen peroxide and streptavidin proteins.

Analyzing the Chiral Purity of Pharmaceuticals: The Application of Cyclodextrin-Based Chiral Selectors in Capillary Electrophoresis

The current review provides a focused analysis of the application of capillary electrophoresis (CE) techniques to determine the chiral purity of pharmaceuticals, with a specific emphasis on cyclodextrin- (CD) based chiral selectors (CSs), highlighting advancements, methodologies, and trends in this area as reported in studies published from 2010 to 2024. The review emphasizes CE’s evolution as a critical tool in this field, discussing its advantages, such as high efficiency, flexibility, relatively low costs, and minimal environmental impact, which make it well-suited for modern pharmaceutical applications. Additionally, it underscores the importance of CE in meeting stringent regulatory requirements for chiral drug substances. A significant shift in method optimization has occurred in the last ten years, shifting from the traditional One-Factor-at-a-Time (OFAT) strategy to the Design-of-Experiments (DoE) approach; this shift has enabled more systematic and robust method development. Furthermore, a common trend in recent years is the application of Quality-by-Design (QbD) principles in method development and optimization, ensuring higher reliability and efficiency. Additionally, there is an increasing focus on developing CE methods capable of detecting both achiral and chiral impurities simultaneously, which enhances the comprehensiveness of the analysis. This review seeks to guide future research and development in optimizing CE methodologies for pharmaceutical applications.

Isolation and purification of DNA double-strand break repair intermediates for understanding complex molecular mechanisms.

Branched DNA molecules are key intermediates in the molecular pathways of DNA replication, repair and recombination. Understanding their structural details, therefore, helps to envisage the mechanisms underlying these processes. While the configurations of DNA molecules can be effectively analysed in bulk using gel electrophoresis techniques, direct visualization provides a complementary single-molecule approach to investigating branched DNA structures. However, for microscopic examination, the sample needs to be free from impurities that could obscure the molecules of interest, and free from the bulk of unwanted non-specific DNA molecules that would otherwise dominate the field of view. Additionally, in the case of recombination intermediates, the length of the DNA molecules becomes an important factor to consider since the structures can be spread over a large distance on the chromosome in vivo. As a result, apart from sample purity, efficient isolation of large-sized DNA fragments without damaging their branched structures is crucial for further analysis. These factors are illustrated by the example of DNA double-strand break repair in the bacterium E. coli. In E. coli recombination intermediates may be spread over a distance of 40 kb which constitutes less than 1% of the 4.6 Mb genome. This study reveals ways to overcome some of the technical challenges that are associated with the isolation and purification of large and complex branched DNA structures using E. coli DNA double-strand break repair intermediates. High-molecular weight and branched DNA molecules do not run into agarose gels subjected to electrophoresis. However, they can be extracted from the wells of the gels if they are agarose embedded, by using β-agarase digestion, filtration, and concentration. Furthermore, a second round of gel electrophoresis followed by purification is recommended to enhance the purity of the specific DNA samples. These preliminary findings may prove to be pioneering for various single-molecule analyses of large and complex DNA molecules of DNA replication, repair and recombination.

Detection of DNA Breaks in Dividing Human Cells by Neutral Comet Assay.

DNA replication is constantly challenged by a wide variety of endogenous and exogenous stressors that can damage DNA. Such lesions encountered during genome duplication can stall replisomes and convert replication forks into double-strand breaks. If left unrepaired, these toxic DNA breaks can trigger chromosomal rearrangements, leading to heightened genome instability and an increased likelihood of cellular transformation. Additionally, cancer cells exhibit persistent replication stress, making the targeting of replication fork vulnerabilities in tumor cells an attractive strategy for chemotherapy. A highly versatile and powerful technique to study DNA breaks during replication is the comet assay. This gel electrophoresis technique reliably detects the induction and repair of DNA breaks at the single-cell level. Herein, a protocol is outlined that allows investigators to measure the extent of DNA damage in mitotically dividing human cells using fork-stalling agents across multiple cell types. Coupling this with automated comet scoring facilitates rapid analysis and enhances the reliability in studying induction of DNA breaks.

Antimalarial analysis of pharmaceutical formulations and biological samples by capillary electrophoresis: the state of the art and applications.

Malaria is a serious public health problem, being an endemic disease in 84 countries, mainly in Africa. This review explores the application of capillary electrophoresis (CE) techniques for analyzing antimalarial drugs, highlighting methods from 2000 to 2023 for the analysis of pharmaceutical formulations and human biological samples. The versatility, selectivity, high efficiency, cost-effectiveness, and high analytical frequency of CE techniques have become attractive choices for pharmaceutical analysis, focusing on quality control and impurity analysis applications. The evolution of achiral and chiral electromigration methods has been described based on the features of each mode of separation: capillary zone electrophoresis (CZE), micellar electrokinetic chromatography, microemulsion electrokinetic chromatography, and capillary electrochromatography. As expected, CZE is reported in most articles owing to its compatibility with drug properties and separation mode. However, it is necessary to perform other separation modes for a few drugs that are present in neutral form. After exhaustive research using different databases and statistical analyses, 27 articles using CE techniques for antimalarial drug analysis were found and are mentioned in this review.

Comparative analysis of electrophoresis and interferometric optical detection method for molecular weight determination of proteins

Analytical detection methods play a pivotal role in scientific research, enabling the identification and quantification of specific analytes in various disciplines. This scientific report aims to compare two very different methodologies for determining the Molecular Mass (MM, also known as Molecular Weight, MW) of proteins: electrophoresis gel and the Interferometric Optical Detection Method (IODM). For this purpose, several proteins with different MM were selected.The electrophoresis technique was employed to validate the structure and MM of different parts or fragments of the Matrix Metallopeptidase 9 antibody (anti-MMP9), antibody against S100 calcium binding protein A6 (anti-S100A6) and Cystatin S4 antibody (anti-CST4) by examining the presence of bands with expected sizes. The IODM was applied to study the above-mentioned proteins (part of the antibodies) together with the protein G, as a reference to correlate the MM and protein sizes with the measured signal.We report the evidence of IODM as a competitive analytical approach for the determination of the MM of proteins for the first time. This innovative method allows for accurate MM determination using minimal sample volumes and concentrations, employing a simple experimental procedure that eliminates the requirement for protein denaturation.

The Pre- and Post-Column Derivatization on Monosaccharide Composition Analysis, a Review.

Polysaccharides, as common metabolic products in organisms, play a crucial role in the growth and development of living organisms. For humans, polysaccharides represent a class of compounds with diverse applications, particularly in the medical field. Therefore, the exploration of the monosaccharide composition and structural characteristics of polysaccharides holds significant importance in understanding their biological functions. This review provides a comprehensive overview of extraction methods and hydrolysis strategies for polysaccharides. It systematically analyzes strategies and technologies for determining polysaccharide composition and discusses common derivatization reagents employed in further polysaccharide studies. Derivatization is considered a fundamental strategy for determining monosaccharides, as it not only enhances the detectability of analytes but also increases detection sensitivity, especially in liquid chromatography (LC), capillary electrophoresis (CE), and gas chromatography (GC) techniques. The review meticulously examines pre-column and post-column derivatization techniques for monosaccharide analysis, categorizing them based on diverse detection methodologies. It delves into the principles and distinctive features of various derivatization reagents, offering a comparative analysis of their strengths and limitations. Ultimately, the aim is to provide guidance for selecting the most suitable derivatization approach, taking into account the structural nuances, biological functions, and reaction dynamics of polysaccharides.

Selectively Isolation and Molecular Identification by dnaJ Gene of Legionella pneumophila Found in Various Water Sources in Dhaka, Bangladesh

Abstract BACKGROUND: Legionella spp. are the etiological agents of Legionnaires’ disease, often severe pneumonia. After the first severe outbreak of Legionella spp., it was found widely dispersed in natural water sources and humid environments and has become a concern for these sources’ water quality and safety. The fecal indicators used for water quality assessment do not predict the presence of these pathogens; thus, in the absence of targeted analysis, the risk is unobserved until the emergence of infections. There is no study on the presence of Legionella in unattended water bodies in Bangladesh, such as swimming pools, air-conditioned cooling water, and stagnant circulatory water systems. METHODS: A total of 42 water samples were collected from 12 different sites in Dhaka, Bangladesh. Acid, heat, and chlorine treatment were applied to reduce the load of non-Legionellae in water samples. Chlorine treatment was used to kill unwanted organisms in water samples. After successful pretreatment, samples were cultured on buffered charcoal yeast extract agar (BCYE) media. Optimum pH, concentrations of chlorine, and temperature were set up by extensive trial and error culturing of Legionella on BCYE media to minimize other bacterial growth and maximize Legionella growth. Isolated colonies were confirmed primarily by morphological characteristics on BCYE media. Isolates were then confirmed by polymerase chain reaction and gel electrophoresis techniques. We detected the presence of 16S rDNA for Legionella spp. and the dnaJ gene for Legionella pneumophila species identification. Antibiotic sensitivity tests (minimum inhibitory concentration) were also conducted to see if it was alarming for our community’s health. RESULTS: Most of the Legionella spp. could grow at various temperatures, such as 45°C, 50°C, and 55°C, and they could also survive at the normal levels of chlorination, such as 0.25 mg/L and 0.5 mg/L. We have found that, out of 12 different sites, 4 out of 5 (80%) isolates from cooling tower water, 1 out of 4 (25%) from the swimming pool, and 1 out of 3 (33%) household stagnant water are contaminated with Legionella. However, 3 out of 5 isolates found are L. pneumophila from cooling water sites. Furthermore, strains of Legionella spp. were resistant to many antibiotics, such as ciprofloxacin, azithromycin, and tetracycline. CONCLUSIONS: Serious and fatal Legionella spp. (L. pneumophila) infections may be transmitted because of the widespread presence of these water sources and the large number of users in the community. This threat is worsened by the survival of Legionella under various commonly used antibiotic conditions and further exacerbated by their antibiotic resistance. Health-care professionals may face great challenges due to the lack of studies about its prevalence and treatment procedures.

Crecimiento micelial y producción de lacasa y peroxidasas por Pleurotus ostreatus y Agrocybe aegerita en fermentación líquida

Mycelial growth of Pleurotus ostreatus and Agrocybe aegerita in malt extract broth was evaluated. Laccase, manganese peroxidase (MnP), lignin peroxidase (LiP) and unspecific peroxygenase (UnP) activities were determined in both strains, using biochemical and polyacrylamide gel electrophoresis techniques. No strain showed MnP, LiP, and UnP activities. P. ostreatus had higher biomass production and laccase yield values than those shown by A. aegerita. P. ostreatus produced approximately twice as much biomass (Xmax) compared to A. aegerita (9.12 and 5.27 g/L, respectively). Maximum laccase activity (Emax) was nine-fold higher in P. ostreatus than in A. aegerita (106.5 and 12.23 U/L, respectively). P. ostreatus and A. aegerita showed 3 bands (estimated to be approximately 42, 55, 85 and 212 kDa) and one band (estimated to be approximately 35.2 kDa) with laccase activity, respectively. These results show that laccases are constitutively produced in both fungal species, whereas MnP, LiP and UnP are inducible enzymes. P. ostreatus has a great potential in the biotechnological production of laccases.

Exploring a new format of thermophoretic measurement – Stop-flow microscale thermophoresis in the narrow-bore transparent capillary

The classic format of the microscale thermophoresis technique (MST) requires the use of disposable glass capillaries. Their total capacity significantly exceeds the volume of the sample used directly in the thermophoretic measurement. An alternative approach that allows for reducing the consumption of sample, reagents and elimination of waste is the use of narrow-bore capillaries, commonly applied in the capillary electrophoresis technique (CE), in combination with injecting sample segment of a reduced length. This article presents the first-ever use of a transparent and flexible narrow-bore capillary for this purpose, which is much easier to handle than a regular non-transparent silica capillary. In this approach, it is possible to perform thermophoretic analysis with a small sample volume, which is delivered to the detection site using pressure-induced flow, or electrophoretically – while simultaneously performing electrophoretic separation. We focused primarily on the former approach, called Transparent Capillary Stop-Flow MicroScale Thermophoresis (TCSF-MST). Its potential was demonstrated in systems presenting the original use of the thermophoresis phenomenon: monitoring the progress of the fluorescent derivatization reaction, determining the critical micellization concentration of a surfactant, and quantitative analysis of fluorophores based on the internal standard method. The obtained results suggest that the new format can be considered an interesting alternative to the traditional MST methodology, and also seems consistent with the idea of “​​green” and “white” analytical chemistry.

Protein Deposition on Sport Mouthguards and the Effectiveness of Two Different Cleaning Protocols.

Objective: To determine which salivary proteins adhere onto sport mouthguards, and to evaluate the effectiveness of different cleaning strategies in removing deposited protein. Methods: Fifteen healthy volunteers used a mouthguard for 1 h. The deposited salivary proteins were analyzed using gel electrophoresis and Western blotting techniques and compared with the protein composition of unstimulated saliva. In addition, the effectiveness of two different cleaning strategies to remove proteins from the mouthguards were compared: rinsing the mouthguards after use with cold tap water and cleaning the mouthguard with a soluble effervescent tablet. Results: Gel electrophoresis showed deposition of proteins of 50-60 kDa and 14 kDa on the mouthguards used in the mouth for 1 h. Western blotting identified these bands as amylase and lysozyme, respectively. Rinsing the mouthguard with cold tap water after use removed 91% of the total amount of deposited proteins, while cleaning with an effervescent tablet removed 99%. Conclusions: During the use of mouthguards, salivary proteins are deposited on their surface. Because salivary proteins can potentially affect bacterial adhesion to mouthguards, proper cleaning after use is recommended. Cleaning the mouthguard with cold tap water or using an effervescent tablet both seem to be effective strategies to remove proteins deposited on sport mouthguards.

Electrophoresis-Based Approach for Characterizing Dendrimer-Protein Interactions: A Proof-of-Concept Study.

Improving the clinical translation of nanomedicine requires better knowledge about how nanoparticles interact with biological environments. As researchers are recognizing the importance of understanding the protein corona and characterizing how nanocarriers respond in biological systems, new tools and techniques are needed to analyze nanocarrier-protein interactions, especially for smaller size (<10 nm) nanoparticles like polyamidoamine (PAMAM) dendrimers. Here, we developed a streamlined, semiquantitative approach to assess dendrimer-protein interactions using a nondenaturing electrophoresis technique combined with mass spectrometry. With this protocol, we detect fluorescently tagged dendrimers and proteins simultaneously, enabling us to analyze when dendrimers migrate with proteins. We found that PAMAM dendrimers mostly interact with complement proteins, particularly C3 and C4a, which aligns with previously published data, verifying that our approach can be used to isolate and identify dendrimer-protein interactions.

Utilizing deep ocean water in yeast fermentation for enhanced mineral-rich biomass production and fermentative regulation by proteomics modulation

Deep Ocean Water (DOW) is rich in minerals and serves as a natural source of nutrients. However, due to the inorganic nature of these minerals, cultivating yeast in DOW could aid in the fermentation process, and simultaneously, the yeast can assimilate the minerals from DOW, resulting in a mineral-enriched yeast biomass. Focusing on three DOW sources off the eastern coast of Taiwan (TT-1, HL-1, HL-2), we fermented various yeast strains of Saccharomyces cerevisiae. Therefore, this study investigates the effects of DOW on yeast growth, alcohol dehydrogenase activity, and the biological absorption of mineral ions by the yeast. Additionally, this research employs two-dimensional electrophoresis techniques to examine how the absorbed minerals influence the regulation of yeast proteins, thereby affecting biomass and metabolism. In the result, S. cerevisiae BCRC 21689 demonstrated a remarkable ability to bio-absorb minerals such as magnesium, calcium, potassium, and zinc from DOW, enhancing its growth and fermentation performance. Proteomic analysis revealed significant shifts in the expression of 21 proteins related to glycolytic and energy metabolism, alcohol metabolism, and growth regulation, all influenced by DOW's mineral-rich environment. This indicates that DOW's mineral content is a key factor in upregulating essential enzymes in glycolytic metabolism and alcohol dehydrogenase. An increase in proteins involved in synthesis and folding processes was also observed, leading to a substantial increase in yeast biomass. This study underscores the potential of DOW as a natural enhancer in yeast fermentation processes, enriching the yeast with diverse minerals and modulating proteomic expression to optimize yeast growth and fermentation.

Angiotensin-converting enzyme I/D gene polymorphism and risk of type 2 diabetes mellitus in Iranian individuals

This study aimed to examine the relationship between type 2 diabetes mellitus (T2DM) and renin-angiotensin system gene polymorphisms in the Iranian population. Additionally, we performed a straightforward meta-analysis of the present articles to better understand this role. A total of 100 Iranian individuals, 50 patients with T2DM, and 50 age-matched healthy individuals were included in this study. DNA was extracted using the salting-out approach, polymerase chain reaction was used to amplify the angiotensin-converting enzyme (ACE) gene, electrophoresis techniques were used, and genotyping was performed. We also searched PubMed, Web of Science, Scopus, and Google Scholar databases for papers published in 2023. We found a significantly higher frequency of I/D genotype in the patient group than in the control group, and the risk of T2DM was 10 times higher in individuals with the I/D genotype (OR, 10 [95% CI, 3.7 to 27]; p < 0.0001) and also 2.85 time higher in individuals with the D allele OR, 2.85 [95% CI, 1.55 to 5.24]; p < 0.001). The ACE polymorphism alleles D and I/D genotypes may increase the risk of developing T2DM in an Iranian population.

Biochemical Changes in Embryogenic and Nonembryonic Callus of Glycyrrhiza glabra L. during Somatic Embryogenesis

The continuous extraction of glycyrrhizin from Glycyrrhiza glabra L. has led to the critical endangerment of this therapeutic plant. Glycyrrhizin, a triterpenoid saponin, is a vital secondary metabolite renowned for its diverse pharmacological benefits, including anti-inflammatory, immunomodulatory, antiallergic, antiulcer properties, and even efficacy against HIV. However, the relentless harvesting of roots for high-value glycyrrhizin results in the obliteration of the entire plant, posing a severe threat to its existence and causing a detrimental impact on biodiversity. To counteract this challenge and conserve the species, sustainable approaches are imperative. One such approach involves the utilization of in vitro regeneration through somatic embryogenesis for clonal proliferation and genetic modification. This report presents findings on SE-associated proteins in Glycyrrhiza glabra L., accomplished through the implementation of the discontinuous gel electrophoresis technique. The study focused on a comparative isozyme analysis employing non-embryogenic and embryogenic calli of G. glabra L. Remarkably, these isozymes were exclusively identified and exhibited affiliations with a spectrum of cellular functions. A substantial portion of these proteins are engaged in a diverse range of metabolic and stress-related functions. This study utilized crude enzyme/total protein extracts as a basis to evaluate the genetic stability of in vitro cultivated plant tissue. Acid phosphatase, esterase, and peroxidase were selected as indicators for this assessment. Intriguingly, esterase and acid phosphatase exhibited significant polymorphism between the segregated embryogenic callus (EC) and non-embryogenic callus (NEC) derived from the same G. glabra L. genotype. Comparative analyses encompassing esterase, acid phosphatase, and peroxidase activities, alongside the expression profiles of these isozymes throughout somatic embryogenesis (SE), were conducted using polyacrylamide gel electrophoresis. Notably, the isoenzyme patterns of calluses were assessed. The activity of isozymes within an embryogenic line of callus surpassed that within a non-embryogenic counterpart. However, no conspicuous alterations in quality were observed between the non-embryogenic and embryogenic calluses.

Design, Synthesis, Structural Investigation and Photo Induced Biological Investigations of Co(II), Ni(II) and Cu(II) Complexes Derived from N,O Donor Schiff Bases.

A series of chelated metal complexes, [Co(LI)2] (1), [Ni(LI)2] (2), [Cu(LI)2] (3) [Co(LII)2] (4), [Ni(LII)2] (5) and [Cu(LII)2] (6) were designed and synthesized from newly synthesized Schiff bases, LI = 2-((E)-(5-(4-fluorophenyl)isoxazol-3-ylimino)methyl)-5-methylphenol and LII = 2-((E)-(5-(4-fluorophenyl)isoxazol-3-ylimino)methyl)-4-chlorophenol. The synthesized compounds were characterized by elemental analysis, nuclear magnetic resonance spectroscopy (NMR), electronic spectroscopy (UV-Vis), infrared spectroscopy (FT-IR), magnetic susceptibility (µeff), electron spin resonance spectroscopy (ESR), Thermogravimetric analysis (TGA), scanning electron microscopy (SEM), energy dispersive X-ray analysis (EDX), and powder X-ray diffraction analysis (P-XRD). The spectral investigations have been clearly suggested 1:2 (metal: ligand) stoichiometric complexes with square planar geometrical arrangement around the metal ion. The thermal gravimmetric analysis (TGA) of these complexes indicates greater thermal stability and various steps involved in thermal decomposition of metal complexes. The binding ability between these metal complexes and calf thymus DNA (CT-DNA) was investigated by UV-Vis, fluorescence spectroscopy and viscometric experiments, which disclosed that, the complexes interacted to CT-DNA via an intercalation binding mode. The cleavage property of metal complexes against pBR322 DNA has been explored by gel electrophoresis technique mediated by UV-illumination and H2O2, showed momentous cleavage activity. Antioxidant activity of all complexes was determined by DPPH free radical scavenging experiment and showed prominent antioxidant activity. Further, the antibacterial and antifungal activities of all compounds were screened against bacterial and fungal strains via in-vitro disc diffusion method. These studies revealed that the complexes showed comparatively more antimicrobial activity than free ligands against tested microbial strains.

Effect of Cu(II) compound containing dipicolinic acid on DNA damage: a study of antiproliferative activity and DNA interaction properties by spectroscopic, molecular docking and molecular dynamics approaches

A polymeric compound formulized as [Cu(µ-dipic)2{Na2(µ-H2O)4]n.2nH2O (I), where dipic is 2,6-pyridine dicarboxylic acid (dipicolinic acid, H2dipic), was synthesized by sonochemical irradiation. The initial in-vitro cytotoxic activity of this complex compared with renowned anticancer drugs like cisplatin, versus HCT116 colon cell lines, shows promising results. This study investigated the interaction mode between compound (I) and calf-thymus DNA utilizing a range of analytical techniques including spectrophotometry, fluorimetry, partition coefficient analysis, viscometry, gel electrophoresis and molecular docking technique. The results obtained from experimental methods reveal complex (I) could bind to CT-DNA via hydrogen bonding and van der Waals forces and the theoretical methods support it. Also, complex (I) indicates nuclease activity in the attendance of H2O2 and can act as an artificial nuclease to cleave DNA with high efficiency. Communicated by Ramaswamy H. Sarma

Impact of Micronutrient Profiles on the Pathological Features and Clinical Outcomes of Colorectal Tumors with Microsatellite Instability

We assessed the role of nutritional factors in conjunction with pathological and clinical characteristics of colorectal tumors with microsatellite instability. Data from 1,135 patients diagnosed with colorectal cancer were analyzed, employing immunohistochemistry for mismatch repair protein detection and polymerase chain reaction-capillary electrophoresis for microsatellite instability gene detection. To explore potential correlations with microsatellite instability status, we evaluated patients’ dietary habits and micronutrient levels, particularly folate and selenium. We discovered that 10.84% of the patients had mismatch repair protein deletions, with MLH1 and PMS2 double deletions being the most prevalent. Notable differences were identified in serum carcinoembryonic antigen levels, tumor location, tumor-node-metastasis stage, tumor differentiation, mucous composition, gross type, size, nodules, nerve invasion, and lymph node ratio between deficient mismatch repair and proficient mismatch repair patients. The deficient mismatch repair group had lower carcinoembryonic antigen levels, a higher incidence of right-sided colon cancer, earlier tumor-node-metastasis stage, poorer differentiation, more mucinous components, larger tumors, and fewer nodules and nerve invasions. High concordance (98.59%) was observed between immunohistochemistry and polymerase chain reaction-capillary electrophoresis techniques in microsatellite instability detection, underscoring the reliability of these methods. Integrating the nutritional parameters with conventional clinical assessments offers a nuanced understanding of the heterogeneity of colorectal cancer, with significant implications for diagnosis and treatment strategies.

Isolation and Identification of dsRNA Mycovirus from Cultivated Mushroom Lentinula edodes

Viruses that infect the edible mushroom Lentinula edodes have been documented to exhibit either disease expression or remain asymptomatic. Nonetheless, our knowledge regarding viruses infecting L. edodes, as well as their diversity, remains relatively limited. This research aimed to isolate double-stranded RNA (dsRNA) molecules from 25 economically significant mushrooms using dsRNA extraction and gel electrophoresis techniques. The results revealed the presence of MSU16 dsRNA segments with a size of approximately 10,000 bp in a Lentinula sp. mushroom sample. This substantial dsRNA suggested the possible identification of the virus within the group of Lentinula edodes mycoviruses. Characterization of the MSU16 dsRNA molecule using a 2-step RT-PCR technique, DNA sequencing, and phylogenetic tree analysis based on partial RdRp nucleotide and amino acid sequences indicated a close relationship to other mushroom viruses found in Lentinula sp., with similarity ranging from 39.4 to 64.6 %. Molecular classification of the ITS gene sequence of the dsRNA-detected MSU16 showed a 92 % identity with L. edodes. However, the mushroom MSU16 could not be classified based on the limited LSU gene sequence available in GenBank. This study suggests that the MSU16 dsRNA molecule represents a Lentinula edodes dsRNA mycovirus isolate from Thailand. Our findings contribute valuable information about mycoviral presence in cultivated edible mushrooms. HIGHLIGHTS An approximately 10,000 bp dsRNA segment was successfully isolated from the Lentinula edodes mushroom isolate MSU16. This dsRNA was characterized through a 2-step RT-PCR, DNA sequencing, and phylogenetic analysis, focusing on partial RdRp nucleotide and amino acid sequences. The analysis indicated a close relationship between the isolated dsRNA and other viruses known to infect L. edodes mushrooms. Furthermore, the ITS gene sequence of MSU16 exhibited a high similarity to the corresponding ITS sequences in L. edodes. GRAPHICAL ABSTRACT

Application of thin boron deposit by electrophoresis as neutron detectors

Abstract Detecting nuclear radiation presents a distinctive challenge, particularly with neutrons, which are neutral particles. The method of direct detection involves the utilization of a converter material, acting as an intermediary. Boron plays a pivotal role in this process, reacting with thermal neutrons to generate alpha particles and lithium, with a notable energy release of 2.314 MeV during the 10B (n,α) 7Li reaction. This facilitates effective identification and measurement of neutrons in radiation detection systems. The paths of the particles α (for E = 1.474 MeV) and Li (for E Li = 0.842 MeV). The active medium of the nuclear detector, typically a gas, undergoes ionization by these highly charged particles, or they form ion pairs that are subsequently collected by electrodes to produce the signal at the detector’s output. Various deposit methods can be used for this purpose, electrophoresis offers a distinct advantage in terms of both simplicity and precision. This study details the utilization of the electrophoresis technique for the deposition of boron on the tube walls of prototype detectors developed within our laboratory.