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Even though dilute (unentangled) polymer solutions cannot act as gel-like sieving media, it has been shown that they can be used to separate DNA molecules in capillary electrophoresis. The separation then comes from sporadic, independent DNA–polymer collisions. We study polymer–polymer collisions in nanochannels (i.e., channels that are smaller than the normal size of the polymers), a situation where a polyelectrolyte is forced to migrate "through" isolated uncharged molecules during electrophoresis. We use Langevin dynamics simulations to explore the nature of these collisions and their effect on the net motion of the two polymer chains. We identify several types of collisions, including some that are unique to nanochannels. When the uncharged polymer is much larger than the polyelectrolyte, the system is reminiscent of gel electrophoresis, and we propose a modified empirical reptation model to explain the data, with an orientation factor that depends on the tube diameter. We also observe that the duration of a collision is a non-monotonic function of the polymer size ratio when the two chains are of comparable size, a surprising resonance-like phenomenon, which, combined with the asymmetric nature of molecular conformations during collision, suggests possible ratchet-like mechanisms that could be used to sort polyelectrolytes in nanodevices. [ABSTRACT FROM AUTHOR]

To determine the evolution of microbial community and microbial shift under anaerobic processes, this study investigates the use of denaturing gradient gel electrophoresis (DGGE). In the DGGE, short- and medium-sized DNA fragments are separated based on their melting characteristics, and this technique is used in this study to understand the dominant bacterial community in mesophilic and thermophilic anaerobic digestion processes. Dairy manure is known for emitting greenhouse gases (GHGs) such as methane, and GHG emissions from manure is a biological process that is largely dependent on the manure conditions, microbial community presence in manure, and their functions. Additional efforts are needed to understand the GHG emissions from manure and develop control strategies to minimize the biological GHG emissions from manure. To study the microbial shift during anaerobic processes responsible for GHG emission, we conducted a series of manure anaerobic digestion experiments, and these experiments were conducted in lab-scale reactors operated under various temperature conditions (28 °C, 36 °C, 44 °C, and 52 °C). We examined the third variable region (V3) of the 16S rRNA gene fingerprints of bacterial presence in anaerobic environment by PCR amplification and DGGE separation. Results showed that bacterial community was affected by the temperature conditions and anaerobic incubation time of manure. The microbial community structure of the original manure changed over time during anaerobic processes, and the community composition changed substantially with the temperature of the anaerobic process. At Day 0, the sequence similarity confirmed that most of the bacteria were similar (>95%) to Acinetobacter sp. (strain: ATCC 31012), a Gram-negative bacteria, regardless of temperature conditions. At day 7, the sequence similarity of DNA fragments of reactors (28 °C) was similar to Acinetobacter sp.; however, the DNA fragments of effluent of reactors at 44 °C and 52 °C were similar to Coprothermobacter proteolyticus (strain: DSM 5265) (similarity: 97%) and Tepidimicrobium ferriphilum (strain: DSM 16624) (similarity: 100%), respectively. At day 60, the analysis showed that DNA fragments of effluent of 28 °C reactor were similar to Galbibacter mesophilus (strain: NBRC 10162) (similarity: 87%), and DNA fragments of effluent of 36 °C reactors were similar to Syntrophomonas curvata (strain: GB8-1) (similarity: 91%). In reactors with a relatively higher temperature, the DNA fragments of effluent of 44 °C reactor were similar to Dielma fastidiosa (strain: JC13) (similarity: 86%), and the DNA fragments of effluent of 52 °C reactor were similar to Coprothermobacter proteolyticus (strain: DSM 5265) (similarity: 99%). To authors knowledge, this is one of the few studies where DGGE-based approach is utilized to study and compare microbial shifts under mesophilic and thermophilic anaerobic digestions of manure simultaneously. While there were challenges in identifying the bands during gradient gel electrophoresis, the joint use of DGGE and sequencing tool can be potentially useful for illustrating and comparing the change in microbial community structure under complex anaerobic processes and functionality of microbes for understanding the consequential GHG emissions from manure.

BACKGROUND RESULTS CONCLUSION The effectiveness of whey obtained by either enzyme (sweet) or acid treatment on wound healing remains unclear. This study investigated the effectiveness of camel and bovine whey prepared enzymatically (CSW and BSW) or by pH reduction (CAW and BAW). After removing the cream from milk, HCl or rennet was used to remove casein, resulting in acid or sweet whey, respectively, followed by lactose removal using dialysis. Casein removal was verified by sodium dodecyl sulfate–polyacrylamide gel electrophoresis. Wound‐healing activity was measured in vitro on HT‐29 cells by scratch assay. All four whey samples (0–1000 mg L−1) were applied on the cells, and the closure of the cell‐free scratched areas was monitored for 48 h.All whey samples increased the cell migration significantly (P < 0.05) to help close the cell‐free areas as an indication of wound healing compared to the negative control. However, the closure amounts between the highest dose (1000 mg L−1) and the control were not significantly different (P > 0.05). Acid whey samples significantly (P < 0.05) elevated the closure speed compared to the sweet whey samples. The highest closure percentage (64.69%) was achieved after treatment with 10 mg L−1 CAW for 48 h. Between the sweet whey samples, BSW was significantly (P < 0.05) more effective in closing the cell‐free zone compared to CSW.This study investigated the wound‐healing potential of camel and bovine whey in vitro by comparing their effects on HT‐29 cell migration. CAW showed the greatest activity and may find uses as a treatment agent in wound healing. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

BACKGROUND RESULTS CONCLUSION Transglutaminase (TGase) is a heat‐resistant biocatalyst with strong catalytic activity, which functions effectively under moderate temperature and pH conditions, and is used widely in protein cross‐linking and recombination. Transglutaminase cross‐linking is a novel and specific modification method for black bean protein isolate (BBPI). This article investigates the effect of transglutaminase cross‐linking on the structure and emulsification performance of heated BBPI.Sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) analysis showed that heated BBPI with TGase had a higher molecular weight than heated BBPI without TGase, and the protein bands widened with increasing enzyme activity, indicating that TGase cross‐linking promoted protein molecule aggregation. A high molecular weight polymer can better stabilize the oil–water interface, preventing the emulsion from layering. Fourier transform infrared (FTIR) spectroscopy showed that the α‐helix content decreased from 15.64% to 13.75%, and the β‐sheet content increased from 48.13% to 54.08%. The decrease in α‐helix content and increase in β‐sheet content could make the structure more stable and improve the emulsifying properties of heated BBPI. When TGase was 20 U g−1, the protein emulsification activity index (EAI) reached its highest value of 1.87 m2 g−1, and the emulsification stability index (ESI) value was 0.27 min (P < 0.05); these figures were 0.19 m2 g−1, and 0.07 min higher, respectively, than in the sample without added TGase.In summary, transglutaminase cross‐linking has a positive effect on the structure and emulsification performance of heated BBPI and can be used as an effective method for BBPI modification. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

The pathogenic bacterium Campylobacter jejuni is a major food safety concern as it can form biofilms that increase its survival and infective potential. Biofilms consist of microbial cells and extracellular matrix (ECM), which is made of water and extracellular polymeric substances (EPS), which are critical for structural integrity and pathogenicity. The aim of this study was to optimize a protocol for the isolation of C. jejuni ECM. We employed eight physical and chemical isolation methods to extract and purify ECM, followed by different qualitative and quantitative analyses using gel electrophoresis and spectroscopy. This comprehensive approach enabled the evaluation of ECM composition in terms of polysaccharides, proteins, and extracellular DNA. The isolation methods resulted in different yields and purities of the extracted ECM components. Centrifugation in combination with chemical treatments proved to be most effective, isolating higher concentrations of polysaccharides and proteins. Additionally, extraction with ether solution facilitated the recovery of high-molecular-weight extracellular DNA. Overall, we provide a refined methodology for ECM extraction from C. jejuni. As polysaccharides and proteins participate in biofilm stability and microbial communication, and extracellular DNA participates in genetic exchange and virulence, our study contributes towards a better understanding of the persistence of this pathogen in the food industry. [ABSTRACT FROM AUTHOR]

The challenge with breast cancer is its ongoing high prevalence and difficulties in early detection and access to effective care. A solution lies in creating tailored metal–organic frameworks to encapsulate anticancer drugs, enabling precise and targeted treatment with less adverse effects and improved effectiveness. Zeolitic imidazolate framework‐8 (ZIF‐8) and carboplatin (CP)‐loaded ZIF‐8 were synthesized and characterized using various analytical techniques. High Resolution‐transmission electron microscopy of ZIF‐8 and CP@ZIF‐8 indicates that the particles had a spherical shape and were nanosized. The drug release rate of CP is 98% under an acidic medium (pH 5.5) because of the dissolution of ZIF‐8 into its coordinating ions, whereas 35% in a physiological medium (pH 7.4) with the addition of CP, the high porosity, and pore diameter of ZIF‐8 decrease from 1243 to 1041 m2/g. Breast cancer MCF‐7 cells were shown greater IC50 in CP@ZIF‐8 (15.01 ± 3.03 µg/mL) than free CP (34.98 ± 4.25 µg/mL) in an in vitro cytotoxicity assessment. The cytotoxicity of the CP@ZIF‐8 against MCF‐7 cells was studied using the methylthiazolyldiphenyl‐tetrazolium bromide method. The morphological changes were examined using fluorescent staining (acridine orange–ethidium bromide and Hoechst 33258) methods. The comet assay assessed the DNA fragmentation (single‐cell gel electrophoresis). The results from the study revealed that CP@ZIF‐8 can be used in the treatment of breast cancer. [ABSTRACT FROM AUTHOR]

Lactococcosis is a common bacterial fish disease caused by Lactococcus garvieae, L. petauri and L. formosensis. Although there are different PCR‐based techniques to identify the etiological agent, none of these can differentiate these two bacteria without sequencing PCR‐amplified fragments. In the present study, we developed a multiplex PCR assay for simultaneous detection and differentiation of L. garvieae and L. petauri. The specificity of the primers was validated against the bacterial DNA of the targeted and non‐targeted bacteria. The sizes of the PCR amplicons were obtained as 204 bp for the DUF1430 domain‐containing protein gene of L. garvieae, 465 bp for the Lichenan permease IIC component gene of L. petauri, and 302 bp for the teichoic acid biosynthesis protein F gene of both L. garvieae and L. petauri. The PCR amplicons were clearly separated by agarose gel electrophoresis. The multiplex PCR assay did not produce any amplification products with the DNA of the non‐targeted bacteria. The multiplex PCR detection limits for L. garvieae and L. petauri were 5 and 4 CFU in pure culture and 50 and 40 CFU/g in spiked tissue samples, respectively. It takes less than 2 h from plate‐cultured bacteria and 3 h from tissue samples to get results. In conclusion, the developed multiplex PCR assay is a rapid, specific, accurate, and cost‐effective method for the detection and differentiation of L. garvieae and L. petauri and is suitable to be used for routine laboratory diagnosis of L. garvieae and L. petauri. [ABSTRACT FROM AUTHOR]

Protein is an important component of beer, and its type, content and molecular weight directly affect the quality of beer, especially the foam quality of beer. Different brands of wheat beer and barley beer available in the market were used for this analysis. The differences in protein composition and foam performance between multi-sample barley and wheat beer were analyzed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and high-pressure size exclusion chromatography. Protein significantly influences beer quality, particularly its foam properties. Wheat beer (WB) has 9.52–84.10% more total protein content than barley beer (BB). The primary proteins in both beers are 6.9–20.1 kDa, with WB having 1.04 g/L more of this protein, 60.11% higher than that of BB. It is one of the main different proteins between WB and BB. WB also contains 66.67% more 20.1–32.4 kDa protein compared to BB. This is one of the main differences between WB and BB proteins. Both 6.9–20.1 kDa and 20.1–32.4 kDa proteins enhance beer viscosity and foam properties. Additionally, WB's > 32.4 kDa protein content is 246.67% higher than BB's, significantly improving beer hydrophobicity and foam performance. These protein differences are key factors in the superior foam quality of WB. [ABSTRACT FROM AUTHOR]

Manganese ion homeostasis is vital for bacteria and is achieved via manganese-dependent transcription factors. Manganese mediation of transcription factor attachment to the corresponding oligonucleotide sequences can be investigated, e.g. via electrophoretic mobility shift assays (EMSA). Formation of specific biocomplexes leads to differences in the migration pattern upon gel electrophoresis. Focusing on electrophoresis in the gas-phase, applying a nano electrospray gas-phase electrophoretic mobility molecular analyzer (nES GEMMA) also known as nES differential mobility analyzer (nES DMA), and on transcription factors (MntR proteins) from Bacillus subtilis and Mycobacterium tuberculosis, we took interest in the gas-phase electrophoresis of the corresponding biospecific complexes. We compared nES GEMMA, separating analytes in the nanometer regime (a few to several hundred nm in diameter) in the gas-phase in their native state according to particle size, to EMSA data. Indeed we were able to demonstrate manganese-mediated attachment of MntR to target genomic sequences with both analytical techniques. Despite some inherent pitfalls of the nES GEMMA method like analyte/instrument surface interactions, we were able to detect the target complexes. Moreover, we were able to calculate the molecular weight (MW) of the obtained species by application of a correlation function based on nES GEMMA obtained data. As gas-phase electrophoresis also offers the possibility of offline hyphenation to orthogonal analysis techniques, we are confident that nES GEMMA measurements are not just complementary to EMSA, but will offer the possibility of further in-depth characterization of biocomplexes in the future. [ABSTRACT FROM AUTHOR]

The bovine lungworm, Dictyocaulus viviparus (Bloch, 1782), is highly pathogenic and disease outbreaks can be difficult to predict and manage. Rapid and accurate diagnosis is vital, but without a sensitive diagnostic test this remains challenging in clinical practice. High performance molecular detection tools are therefore required to improve the diagnosis of this parasite and promote the implementation of strategic control measures. Loop-mediated isothermal amplification (LAMP), a rapid DNA assay, offers potential for field-based detection. Here we report a novel LAMP assay (DviLAMP), that was designed to target the D. viviparus internal transcribed spacer 2 (ITS2) ribosomal DNA region. Firstly, genomic DNA was extracted from a single D. viviparus L1 larva to amplify and clone the ITS2 into the recombinant plasmid (DviITS2). The DviLAMP successfully detected the target, with results shown by gel electrophoresis and real-time analysis, in addition to point-of-care amenable end-point detection: colorimetry and lateral flow dipstick (LFD). Analytical sensitivity can detect 0.5 ng DviITS2 following 45 min of incubation at 64°C, increasing to just 1 pg following 90 min of incubation. Using the same primers, other nematodes of cattle, Ostertagia ostertagi and Cooperia oncophora, were also detectable both by gel electrophoresis and real-time. However, when FITC and biotin tagged primers were incorporated to adapt the DviLAMP to LFD end-point detection, the LFD showed specific detection of D. viviparus. Further development of DviLAMP as a point-of-care test could significantly improve the sensitivity of lungworm diagnosis in the field. [ABSTRACT FROM AUTHOR]

The diminishing of food waste is gaining increasing importance, especially in context with a growing population and a need for the sustainable use of food resources. A more precise determination of the best-before date can contribute to this general aim. As proteoforms can be regarded as indicators for ecophysiological influences, their suitability for determining the spoilage and, consequently, the shelf-life of food is suggested. Proteoforms reflect the spoilage of food more accurately. The aim of the present study was to develop an efficient proteomics workflow to determine the shelf-life of milk as a prominent target. In this case, raw milk was chosen as model, as it degrades much faster. The integration of different multivariate analysis techniques was used to analyze the spoilage of raw milk with regard to aspects of its proteome. As the feasibility of such an approach has already been demonstrated in previous studies, it is further necessary to enable a robust and reproducible workflow, primarily gaining appropriate numbers and amounts of peptides when the research question differs and other dairy products are evaluated. In the present study, two approaches for gaining peptides were considered: In addition to a direct hydrolysis of a protein-rich sample solution, in-gel hydrolysis is another common approach in proteomics. By separating the proteins in a traditional gel electrophoresis before hydrolysis, the change in the individual proteins and, consequently, potential peptides can be monitored more specifically during storage. However, the traditional approach offers not only possibilities but also limitations that must be considered. The study showed that it is beneficial to apply a combination of different application strategies, as they complement each other and can thus increase the information content of a sample or confirm a theory. Mass spectrometric features, which represent a chemical–structural change of all kinds of compounds during storage, were selected, and three of them were identified as peptides, originating from α-s1-casein. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

RG1 is a quadruplex‐forming sequence in the SARS‐CoV‐2 genome proposed as possible therapeutic target for COVID‐19. We demonstrate that the dominant conformation of RG1 under physiological conditions differs from the parallel quadruplex previously assumed. Through comprehensive investigations employing CD, UV, NMR, DSC, gel electrophoresis, MD simulations, in silico spectroscopy and the use of truncated RG1 sequences, we have identified this stable conformation as an RNA G‐triplex composed of two G‐triads. We believe this previously unreported RNA structure could serve as a novel therapeutic target. Our findings open new avenues for further studies on the presence and biological role of RNA G‐triplexes in vivo. [ABSTRACT FROM AUTHOR]

Loop-Mediated Isothermal Amplification (LAMP) represents a valuable technique for DNA/RNA detection, known for its exceptional sensitivity, specificity, speed, accuracy, and affordability. This study focused on optimizing a LAMP-based method to detect early signs of Plasmopara halstedii, the casual pathogen of sunflower downy mildew, a severe threat to sunflower crops. Specifically, a set of six LAMP primers (two outer, two inner, and two loop) were designed from P. halstedii genomic DNA, targeting the ribosomal Large Subunit (LSU). These primers were verified by in silico analysis and experimental validation using both target and non-target species' DNAs. Optimizations encompassing reaction conditions (temperature, time) and component concentrations (magnesium, Bst DNA polymerase, primers, and dNTP) were determined. Validation of these optimizations was performed by agarose gel electrophoresis. Furthermore, various colorimetric chemicals (Neutral Red, Hydroxynaphthol Blue, SYBR Safe, Thiazole Green) were evaluated to facilitate method analysis, and the real-time analysis has been optimized, presenting multiple approaches for detecting sunflower downy mildew using the LAMP technique. The analytical sensitivity of the method was confirmed by detecting P. halstedii DNA concentrations as low as 0.5 pg/μl. This pioneering study, establishing P. halstedii detection through the LAMP method, stands as unique in its field. The precision, robustness, and practicality of the LAMP protocol make it an ideal choice for studies focusing on sunflower mildew, emphasizing its recommended use due to its operational ease and reliability. [ABSTRACT FROM AUTHOR]

Background: Comprehensive treatment of gastric cancer (GC) is progressing, but the rapid proliferation and metastasis of GC remains a cause of high recurrence and mortality rates. In this study we investigated GC-associated circRNA tending to yield more insight into the mechanisms of gastric cancer development. Methods: We detected the expression levels of circTSN in GC tissues and cell lines using qRT-PCR. The circular structure of circTSN was confirmed by Sanger sequencing, agarose gel electrophoresis and RNase R. A series of cell functional experiments were employed to investigate the implication of circTSN aberrant expression on the proliferation and metastasis of GC cells. The predicted binding domain between circTSN and miR-1825 was analyzed by luciferase reporter gene analysis. Meanwhile, subcutaneous tumor xenografts in nude mice were used to validate the role of circTSN in vivo. Results: It was found that RNA levels of circTSN were significantly elevated in GC tissues and cell lines, which was also confirmed to contain a closed-loop structure. CCK8, clone formation, EdU, transwell and in vivo experiments indicated that the highly expressed circTSN was involved in the proliferation and metastasis process of GC. In addition, circTSN modulates the expression of SLC38A2 by sequence-specific binding to miR-1825. Conclusion: This study identified that circTSN, which is highly expressed in GC, was able to contribute to the proliferation and metastasis of GC cell through miR-1825/SLC38A2 axis and this might provide a new candidate target for the precision treatment of GC. [ABSTRACT FROM AUTHOR]

Background: Encapsulation of siRNA fragments inside liposome vesicles has emerged as an effective method for delivering siRNAs in vitro and in vivo. However, the liposome's fluid-phospholipid bilayer of liposomes allows siRNA fragments to diffuse out of the liposome, decreasing the dose concentration and therefore the effectiveness of the carrier. We have previously reported that β-cyclodextrins formulated in liposomes help increase the stability of siRNAs in cell culture medium. Here, we continued that study to include α, γ, methyl-β-cyclodextrins and β-cyclodextrin-modified gold and selenium nanoparticles. Methods: We used Isothermal Titration Calorimetry to study the binding thermodynamics of siRNAs to the cyclodextrin-modified nanoparticles and to screen for the best adamantane derivative to modify the siRNA fragments, and we used gel electrophoresis to study the stabilization effect of siRNA by cyclodextrins and the nanoparticles. Results: We found that only β- and methyl-β-cyclodextrins increased siRNA serum stability. Cyclodextrin-modified selenium nanoparticles also stabilize siRNA fragments in serum, and siRNAs chemically modified with an adamantane moiety (which forms inclusion complexes with the cyclodextrin-modified-nanoparticles) show a strong stabilization effect. Conclusions: β-cyclodextrins are good additives to stabilize siRNA in cell culture medium, and the thermodynamic data we generated of the interaction between cyclodextrins and adamantane analogs (widely used in drug delivery studies), should serve as a guide for future studies where cyclodextrins are sought for the delivery and solvation of small organic molecules. [ABSTRACT FROM AUTHOR]

The rapid advancement of biotechnology over the recent decades has amplified the importance of DNA detection and quantification assays. Many of these assays, such as gel electrophoresis, microscopy, flow cytometry, and the detection of amplification in quantitative polymerase chain reaction (qPCR), rely on the use of DNA‐binding fluorescent dyes. This article presents a comprehensive study of six Thiazole‐Orange‐based fluorescent DNA‐binding dyes: SYBR Safe, SYBR Green, Pico Green, SYTO‐16, SYTO‐9, and the benzothiazole‐based analogue (TOPhBu) of the latter. The selected DNA markers were synthesized at a 10‐milligram scale and characterised spectroscopically to quantify their fluorescence enhancement upon binding to double‐stranded DNA. The ability of the dyes to detect DNA at low concentrations was evaluated using two new metrics, absolute fluorescence enhancement (AFE) and relative fluorescence enhancement (RFE). Quantum chemical calculations shed new light on the mechanism of their fluorogenicity through modelling the excited state behaviour and DNA binding of the dyes. Their analytical performance was further tested in qPCR experiments. The experimental results of this work highlight some important differences in the sensitivity and qPCR efficiency of the studied DNA‐binding dyes which will facilitate the DNA marker selection for analytical purposes and the future development of novel DNA sensors. [ABSTRACT FROM AUTHOR]

Summary: Maillard reaction can enhance the functional characteristics of the protein. Herein, we investigated changes in the physicochemical and biological properties of gelatin (GA)‐xylooligosaccharide (XOS) conjugates following Maillard reaction. The results showed that the browning intensity of the conjugates increased with the prolongation of heating time. Additionally, sodium dodecyl sulphate‐polyacrylamide gel electrophoresis revealed some macromolecule polymer formed, and the grafting degree of GA was augmented to 40.05% with prolongation of heating time. GA glycation with XOS decreased α‐Helix content and increased level of β‐Turn level, and exerted a slight impact on contact angle. Moreover, the results of in vitro simulated gastrointestinal digestion analysis showed that the GA‐XOS mixture enhanced resistance to digestion after Maillard reaction for 60 min, affording hydrolysis ratios ranging from 17.86–27.79% and 25.03–35.47% in simulated gastric fluid and intestinal fluid, respectively. The antioxidant ability and heat stability of the conjugates was visibly enhanced following heating treatment. Additionally, increasing the mass of XOS accelerated the antioxidant capacities of GA‐XOS conjugates. The finding will assist the fabrication of GA‐XOS conjugates with highly functional properties by Maillard reaction. [ABSTRACT FROM AUTHOR]

Summary: While dietary components such as phytate reduce calcium absorption, bioactive peptides could potentially enhance calcium absorption. This study aimed to characterise the peptides derived from casein and assess their effect on calcium uptake by Caco‐2 cells. Casein was subjected to hydrolysis by Alcalase and Flavourzyme to produce 4‐h and 16‐h casein hydrolysates. High‐performance liquid chromatography analysis showed that hydrolysis caused significant increases and decreases in the content of some amino acids. Casein hydrolysates achieved 40.36% and 52.64% degree of hydrolysis, as determined by the o‐phthaldialdehyde method. Sodium dodecyl sulphate‐polyacrylamide gel electrophoresis and dynamic light scattering analyses revealed that the peptides had molecular weight less than 15 kDa and diameter between 100 and 1000 nm. The 4‐h casein hydrolysate enhanced calcium uptake by Caco‐2 cells monolayer at low calcium intake, possibly via transcellular absorption. These results provide insight into the nutraceutical potential of casein hydrolysates as enhancer of calcium absorption. [ABSTRACT FROM AUTHOR]

Despite the multiple health benefits of vitexin, a natural flavonoid glycoside, its extremely low solubility, poor absorption, and low bioavailability hinder its effective use. This study attempted to enhance the solubility, antioxidant activity, and bioavailability of vitexin by nanoencapsulation in a delivery system of mung bean protein isolate nanoparticles through the processes of pH shifting and pH shifting combined with ultrasonication. The results showed that the combined method significantly induced protein denaturation and promoted protein-vitexin interactions via hydrophobic interactions, in comparison to pH shifting alone. The vitexin-loaded nanoparticles prepared using the combined method had an average size of 40.99 nm, a homogeneous size distribution (0.267), and a zeta potential of − 31.68 mV. The encapsulation efficiency of the nanoparticles produced through the combined method (88.52%) was significantly higher (p < 0.05) compared to those prepared using pH shifting alone (52.10%). Additionally, the solubility and antioxidant activity of the encapsulated vitexin were markedly increased. High stability of the nanoparticles during dynamic in vitro gastrointestinal digestion was observed by sodium dodecyl sulfate‒polyacrylamide gel electrophoresis. The amount of vitexin remaining in the nanoparticles after digestion was 72.89%, which was available for absorption. In summary, this study successfully enhanced the solubility, antioxidant activity, and bioavailability of vitexin through nanoencapsulation in mung bean protein isolate nanoparticles using a combination of pH shifting and ultrasonication, demonstrating significant improvements over pH shifting alone. [ABSTRACT FROM AUTHOR]

Fluazuron is a novel veterinary pour-on antitick formulation which can be applied simultaneously with bovine reproduction management strategies. Considering the economic importance of the livestock industry in many countries, it is important to know whether antiparasitics such as fluazuron may cause embryonic loss. The aim of this study was to evaluate the toxicological effect of fluazuron on bovine oocytes during in vitro maturation. The best fluazuron concentrations were determined in a preliminary experiment on Chinese hamster ovary (CHO)-K1 cells and further used to compare fluazuron toxicity in both study models. Results of the annexin V and alkaline single cell gel electrophoresis assays demonstrated that fluazuron caused cytotoxicity and genotoxicity in bovine cumulus cells at all the concentrations tested (50, 75 and 100 μg fluazuron/mL). The evaluation of cortical granules and mitochondria distribution showed that cytoplasmic maturation was not affected by fluazuron treatment. However, a decrease in metaphase II + polar body, degenerate oocytes as well as disorganized chromatin in polar body were observed at all concentrations tested. Whereas the fertilization process was not altered by 50 μg/mL fluazuron, the embryo development rate decreased significantly. No significant differences were observed in any of the oxidative stress parameters assessed. This study contributes to a better understanding of fluazuron in bovines, suggesting that the antiparasitic may affect bovine reproduction and might cause embryo loss. • Fluazuron induces genotoxicity in CHO-K1 and genotoxicity and apoptosis in cumulus cells. • Fluazuron reduces metaphase II and polar body (PB) formation, degenerates oocytes, and disorganizes PB chromatin. • Embryo development rate decreased significantly after cumulus-oocyte cells exposure to 50 μg fluazuron/mL. • Fluazuron does not alter the oxidative stress parameters assessed in bovine cumulus cells and oocytes. [ABSTRACT FROM AUTHOR]

Amaranth is a promising staple food that produces seeds with excellent nutritional quality. Although cultivated species intended for grain production have interesting agronomic traits, relatively little is known about wild species, which can prosper in diverse environments and could be a rich genetic source for crop improvement. This work focuses on the proteomic comparison between the seeds of wild and cultivated amaranth species using polarity-based protein extraction and two-dimensional gel electrophoresis. Differentially accumulated proteins (DAPs) showed changes in granule-bound starch synthases and a wide range of 11S globulin isoforms. The electrophoretic profile of these proteins suggests that they may contain significant phosphorylation as post-translational modifications (PTMs), which were confirmed via immunodetection. These PTMs may impact the physicochemical functionality of storage proteins, with potential implications for seed agronomic traits and food system applications. Low-abundant DAPs with highly variable accumulation patterns are also discussed; these were involved in diverse molecular processes, such as genic regulation, lipid storage, and stress response. [ABSTRACT FROM AUTHOR]

Peroxidase is a biotechnologically important enzyme. The purification of peroxidase from the root of Citrullus colocynthis was carried out in a simple two-step process with maximum purity level. The sample was extracted in a high salt buffer, and the enzyme was partially purified with a Q-Sepharose anion exchange column. Final purification was carried out with HighLoad 16/600 Superdex G-75 column. The purified protein was analysed with SDS gel electrophoresis, which suggested a single band of approximately 35 kDa. Further, the enzyme was identified with the help of Mass spectrometric analysis using an ESI-QTOF Mass spectrometer. The study will be helpful for the isolation and its commercial uses in biotechnology. [ABSTRACT FROM AUTHOR]

Copyright of Food Research & Development is the property of Food Research & Development Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Food Research & Development is the property of Food Research & Development Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

In common wheat (Triticum aestivum L.), the protein content and glutenin protein composition are the key quality-determining parameters. Allelic variations, especially in high-molecular-weight glutenin subunits (HMW-GSs), affect bread quality significantly. The HMW-GS Glu-1 locus consists of two tightly linked genes encoding x- and y-type subunits that exhibit highly variable frequencies. In this study, we evaluated Glu-B1 alleles using allele-specific PCR markers in 44 domestic wheat cultivars. The composition of Glu-1Bx7+Glu-1By8 in the 24 cultivars was either Glu-1Bx7+Glu-1By8, Glu-1Bx7*+ Glu-1By8, or Glu-1Bx7*+Glu-1Bx8*. In addition, the two cultivars initially identified Glu1Bx7+Glu-1By8* were corrected to Glu-1Bx7*+Glu-1By8*. Seven cultivars previously classified as having Glu-1Bx7+Glu-1By9 composition contained Glu-1Bx7*+ Glu-1Bx9. The allele composition of the cultivar was identified as Glu-1Bx20+Glu-1By20 instead of Glu-1By20. The HMW-GSs of 21 wheat varieties were analyzed using ultra-performance liquid chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. These results will be helpful for evaluating the composition of Glu-B1 alleles in domestic wheat and accurately assessing the quality of domestic wheat flour. [ABSTRACT FROM AUTHOR]

In connection with the requirements of international and national organizations to comply with ethical principles for experiments using animals, alternative test systems are being sought and tested to replace animals in ecological, toxicological, and genotoxicological studies. One such alternative may be the nematode Caenorhabditis elegans, which has a biotransformation system of chemical compounds similar to the mammalian system. The genotoxicity of the pesticide paraquat and the antibacterial agent furacilin was studied on C. elegans by horizontal gel electrophoresis of total DNA in order to assess its integrity. It has been shown that paraquat in concentrations of 0.01 to 0.05 mol/L and furacilin in concentrations of 0.0001 and 0.00025 mol/L caused DNA breaks in nematode cells. The antioxidant N-acetylcysteine at a concentration of 0.01 mol/L reduced the genotoxicity of both compounds. [ABSTRACT FROM AUTHOR]

Gel electrophoresis and size exclusion chromatography (SEC) are vital techniques in biochemical research, employing gel matrix structures made of polysaccharides or synthetic polymers like polyacrylamide for the analysis and separation of macromolecules. Polysaccharides, such as agarose, offer safer alternatives to acrylamide. Polysaccharide gels, notably agarose, facilitate the analysis and purification of proteins and nucleic acids through a molecular sieving mechanism. Gel electrophoresis for proteins is mainly divided into denaturing and native methods. Denaturing electrophoresis with sodium dodecyl sulfate (SDS) simplifies protein migration but disrupts molecular interactions. Conversely, native gel electrophoresis, without SDS, allows proteins to migrate based on the running pH and the isoelectric point of the proteins, while nucleic acids consistently migrate toward the anode. The electrophoresis of proteins with variable charges presents complexes. This review focuses on the use of polysaccharides, particularly agarose, for native gel electrophoresis, highlighting their applications in separating macromolecules. It also discusses the applications and limitations of agarose gels when used as a matrix for electrophoresis. Such information should help in designing electrophoresis experiments using polysaccharides. [ABSTRACT FROM AUTHOR]

Background: In vitro-transcribed (IVT) mRNA has been established as a promising platform for therapeutics and vaccine development. Double-stranded RNA (dsRNA) is a major impurity of IVT mRNA and can trigger unfavored immune responses, potentially causing adverse events in patients. Existing dsRNA detection and quantitation methods, such as gel electrophoresis, ELISA, or homogeneous time-resolved fluorescence (HTRF), have low sensitivity or are time-consuming. A recently published lateral flow immunoassay (LFSA) was shown to be fast, but it lacks the sensitivity for dsRNA with uridine modifications. Methods: In this study, we provided a possible explanation for the reduced sensitivity of existing quantitation methods for dsRNA with modified uridines by characterizing the binding affinities of commonly used anti-dsRNA antibodies. Then, a rapid and sensitive biolayer interferometry (BLI) dsRNA detection assay utilizing Flock House Virus (FHV) B2 protein was developed to overcome the challenges in dsRNA detection and the reduced sensitivity. Results: This assay allows the detection of dsRNA with different uridine modifications (ψ, m1ψ, 5 moU) with similar sensitivity as dsRNA without modification. Furthermore, we demonstrated this method can be used to quantify both short and long dsRNA, as well as hairpin-structured dsRNA, providing a more comprehensive detection for dsRNA impurities. Moreover, we applied this assay to monitor dsRNA removal through a purification process. Conclusions: Taken together, this BLI method could enable real-time monitoring of impurities in IVT mRNA production to prevent immunogenicity stemming from dsRNA. [ABSTRACT FROM AUTHOR]

R-loops, structures that play a crucial role in various biological processes, are integral to gene expression, the maintenance of genome stability, and the formation of epigenomic signatures. When these R-loops are deregulated, they can contribute to the development of serious health conditions, including cancer and neurodegenerative diseases. The detection of R-loops is a complex process that involves several approaches. These include S9.6 antibody- or RNAse H-based immunoprecipitation, non-denaturing bisulfite footprinting, gel electrophoresis, and electron microscopy. Each of these methods offers unique insights into the nature and behavior of R-loops. In our study, we introduce a novel protocol that has been developed based on a single-molecule DNA combing assay. This innovative approach allows for the direct and simultaneous visualization of RNA:DNA hybrids and replication forks, providing a more comprehensive understanding of these structures. Our findings confirm the transcriptional origin of the hybrids, adding to the body of knowledge about their formation. Furthermore, we demonstrate that these hybrids have an inhibitory effect on the progression of replication forks, highlighting their potential impact on DNA replication and cellular function. [ABSTRACT FROM AUTHOR]

The Ikaros zinc finger factor 1 is a transcription factor with a well-known role in B- and T-cell development. The deletions of IKZF1 have an established significance in acute lymphoblastic leukemia, while reports on its prevalence and prognostic significance among ALL subtypes and regions vary. Breakpoint-specific qPCR is a practical method for testing of the most frequent types of IKZF1 deletions, considering there is clustering of the deletion events. The most commonly reported deletions are Δ4–7, Δ4–8, Δ2–7, and Δ2–8, with deletion Δ4–7 being the most common one. We retrospectively administered a breakpoint-specific qPCR design for screening for the most frequent types of IKZF1 deletions to 78 ALL patients that were diagnosed and treated between 2010 and 2022. We observed the products through gel electrophoresis, and we conducted descriptive statistics, EFS, and OS analyses. Our study found 19 patients with IKZF1 deletions, with two subjects manifesting more than one deletion. The prevalence in the different subgroups was as follows: Ph/+/ B-ALL 46%, Ph/−/ B-ALL 30%, T-ALL/LBL 4%. There was a statistically significant difference in EFS of 39 vs. 0% in favor of patients without deletions (p = 0.000), which translated to a difference in OS of 49 vs. 0% (p = 0.001). This difference was preserved in the subgroup of Ph/−/ B-ALL, while there was no significant difference in the Ph/+/ B-ALL. The most frequently observed type of deletion (15 out of 19) was the Δ4–7. There is a strong negative prognostic impact of the IKZF1 deletions at diagnosis in the observed population. IKZF1 deletion testing through breakpoint-specific qPCR is a practical approach in diagnostic testing for this risk factor. IKZF1 deletions may warrant treatment decisions and intensified treatment strategies to overcome the negative prognostic impact. [ABSTRACT FROM AUTHOR]

GelBox is open-source software that was developed with the goal of enhancing rigor, reproducibility, and transparency when analyzing gels and immunoblots. It combines image adjustments (cropping, rotation, brightness, and contrast), background correction, and band-fitting in a single application. Users can also associate each lane in an image with metadata (for example, sample type). GelBox data files integrate the raw data, supplied metadata, image adjustments, and band-level analyses in a single file to improve traceability. GelBox has a user-friendly interface and was developed using MATLAB. The software, installation instructions, and tutorials, are available at https://campbell-muscle-lab.github.io/GelBox/. NEW & NOTEWORTHY: GelBox is open-source software that was developed to enhance rigor, reproducibility, and transparency when analyzing gels and immunoblots. It combines image adjustments (cropping, rotation, brightness, and contrast), background correction, and band-fitting in a single application. Users can also associate each lane in an image with metadata (for example, sample type). [ABSTRACT FROM AUTHOR]

Banana is one of the primary fruits cultivated in Malaysia and currently decimated by the emergence of a disease, known as banana blood disease (BBD) which caused by Ralstonia syzygii subsp. celebesensis (Rsc). The BBD has significantly affected the yield and profits of the worldwide banana industry. To date, various approaches including chemical and biological controls have been attempted to manage this disease but none of them succeed in controlling the disease. The uses of lactic acid bacteria (LAB) in managing plant diseases have been reported earlier but little information is available. Therefore, this project is designed to identify and investigate potential plant-associated LAB as biological control agent (BCA) against Rsc by using agar well diffusion method. The inhibition zones of each well were observed after 72h and the isolated LAB that showed inhibition zones were proceed for molecular characterization using PCR amplification followed by gel electrophoresis. The sequences were used for phylogenetic analysis. In addition, each of the potential LAB were used to identify their morphological characterizations and biochemical testing. Throughout the study, the highest inhibition zones of LAB from kimchi and fermented milk achieved a diameter of 21.30 mm and 28.70 mm, respectively. Kimchi isolates showed the highest similarity which is 97% as Lactiplantibacillus plantarum species. Among the fermented milk isolates, the highest similarity which is 98% identified as Lacticaseibacillus paracasei. [ABSTRACT FROM AUTHOR]

Therapeutic messenger RNAs (mRNAs) have emerged as powerful tools in the treatment of complex diseases, especially for conditions that lack efficacious treatment. The successful application of this modality can be attributed to its ability to encode entire proteins. While the large nature of these molecules has supported their success as therapeutics, its extended size creates several analytical challenges. To further support therapeutic mRNA development and its deployment in clinical trials, appropriate methods to support their characterization must be developed. In this review, we describe current analytical methods that have been used in the characterization of RNA quality, identity, and integrity. Advantages and limitations from several analytical techniques ranging from gel electrophoresis to liquid chromatography–mass spectrometry and from shotgun sequencing to intact mass measurements are discussed. We comprehensively describe the application of analytical methods in the measurements of capping efficiency, poly A tail analysis, as well as their applicability in stability studies. [ABSTRACT FROM AUTHOR]

A theoretical framework is established for the gel diffusiophoresis of a spherical colloidal particle moving through an uncharged dilute porous polymer gel medium when an electrolyte concentration gradient field is applied. The network of cross-linked polymer segments is treated as a porous skeleton containing an electrolyte solution using the Brinkman–Debye–Bueche model. We derive a general expression for the gel-diffusiophoretic mobility of a charged spherical colloidal particle. Based on this general mobility expression, we farther derive a closed-form approximate expression for the gel-diffusiophoretic mobility of a weakly charged spherical particle correct to the second order of the particle's zeta potential. The obtained mobility expression depends on the Debye–Hückel parameter and the Brinkmann parameter. [ABSTRACT FROM AUTHOR]

Objective: We sought to evaluate accurate and reproducible detection of Myxobolus cerebralis (Mc), the causative agent of whirling disease, by using nested polymerase chain reaction (nPCR) and three previously established real‐time quantitative PCR (qPCR) assays: K18S (Kelley 18S), C18S (Cavender 18S), and Hsp70 (heat shock protein 70). We used a "fit for purpose" approach combined with intra‐ and interlaboratory testing to identify a molecular testing method that would be equivalent to the currently accepted nPCR procedure for Mc. Methods: Assay performance was compared using a combination of intra‐ and interlaboratory testing that used synthetic gBlocks along with naturally and experimentally infected fish tissue. North American isolates representing geographically distinct locations were also tested using all three assays. Result: The K18S and C18S assays exhibited high assay sensitivity, intra‐ and interlaboratory repeatability of sample replicates, and reproducible identification of all test samples across multiple laboratories. In contrast, the Hsp70 assay failed to detect several positive samples at low DNA concentrations during intra‐ and interlaboratory testing. The K18S assay was the only procedure that demonstrated perfect detection accuracy when testing geographically distinct Mc isolates. Results demonstrated the K18S assay is robust under variable test conditions, is more accurate than the C18S and Hsp70 assays, and provides detection capabilities equivalent to those of the currently accepted nPCR confirmation assay "gold standard" that is described in the American Fisheries Society–Fish Health Section (AFS–FHS) Blue Book. Conclusion: The "fit for purpose" approach and preliminary completion of the World Organization for Animal Health validation pathway demonstrate that the K18S assay provides an alternate method for Mc testing. This work provides the foundation for acceptance of the K18S assay into the AFS–FHS Blue Book as a standardized test procedure for Mc. Impact statementThe myxozoan parasite Myxobolus cerebralis (Mc) can cause whirling disease in salmonids, which can negatively impact hatchery and wild populations. Rapid, sensitive detection of developmental stages is important to identify Mc through monitoring and testing of presumptive findings. The current "gold standard" for molecular testing is nested PCR, which requires two consecutive reactions and visualization of amplicons using gel electrophoresis. Alternatively, three previously published real‐time quantitative PCR (qPCR) assays offer a faster, more sensitive approach for Mc testing. During intra‐ and interlaboratory comparisons, we evaluated these qPCR assays using a range of fish tissue and gBlocks. The "fit for purpose" approach and preliminary completion of the World Organization for Animal Health validation pathway demonstrate that the K18S assay provides an alternate molecular testing procedure for Mc. [ABSTRACT FROM AUTHOR]

An accurate and standardized diagnosis is needed to detect microbial resistance and changes in an organism's genome. The main problems faced in detecting Mycobacterium tuberculosis from clinical samples are the quality of the primers and the specificity of a gene target. This study aims to determine the main characteristics when evaluating primers' s fitness generated from in silico experiments using Thermo Fisher Scientific® software and optimization efforts to increase sensitivity to clinical samples, this study also objectively evaluates the potential of atpE as a detection target for MTB using sputum specimens from a clinical setting. A total of 25 clinical samples from pulmonary tuberculosis patients were extracted using the Zymo DNA Extraction® kit and 11 of them were also extracted using the boiling method. DNA isolates were tested for concentration and purity with Nanodrops®. The primer was designed using the open-access software Thermo Fisher Scientific Oligo Primer design tool ®. Reference primer of atpE was obtained from previous research in vitro. Primary validation was carried out using gel electrophoresis. The ROC curve analysis was carried out with GraphPad Prism version 8.4. The atpE primers designed with Thermo Fisher Scientific® showed a 100% detection rate against the positive control bacterial DNA of Mycobacterium tuberculosis H37Rv. The overall atpE primers from Thermo Fisher Scientific® had a sensitivity of 61.54% and a specificity of 100% compared to the reference primers against the clinical sample. In summary, the atpE gene is a good candidate for both species and drug-resistance genes targeted for PCR and genotype profiling in M. tuberculosis. [ABSTRACT FROM AUTHOR]

N‐Methylbenzylamine and ethylenediamine were utilized as primary and auxiliary ligands, respectively, alongside metal salts to synthesize ternary metal complexes: [FeCl3(Nmba)(en)], [CoCl2(Nmba)(en)(H2O)] and [CuCl2(Nmba)(en)(H2O)]. These complexes were thoroughly characterized using spectral and analytical techniques, revealing an octahedral geometry for all. Coats–Redfern calculations indicated their non‐spontaneous nature yet highlighted their thermal stability. DNA binding studies unveiled a groove binding mode for the complexes, with intrinsic binding constants (Kb) and Stern–Volmer quenching constant (Ksq) supporting their strong binding capabilities. Nuclease activity against pBR322 was assessed through gel electrophoresis. Additionally, docking studies using AutoDock 4.2 software provided insights into their binding affinities. In terms of biological activities, the Cu complex demonstrated superior cytotoxicity and antibacterial and antifungal properties compared to the other ternary metal complexes. This suggests its potential for further exploration in biomedical applications. [ABSTRACT FROM AUTHOR]

In this work, dendritic, water‐soluble, and nonaggregated axially substituted silicon phthalocyanines (EA‐TD‐Si and EA‐TD‐SiQ) were synthesized and characterized. Then, the supercoiled pBR322 plasmid DNA damage properties of EA‐TD‐Si and EA‐TD‐SiQ were examined using agarose gel electrophoresis without light irradiation. In addition, the cytotoxicity and phototoxicity of EA‐TD‐Si and EA‐TD‐SiQ on human endometrium adenocarcinoma (HEC‐1B) cells were investigated using an MTT assay. The results indicated that EA‐TD‐SiQ had high photodamage properties via singlet oxygen depending on the light dose, while EA‐TD‐Si did not show any DNA damage without light. In addition, EA‐TD‐SiQ had higher phototoxic effects than EA‐TD‐Si on HEC‐1B cells according to the MTT assay. These results revealed the potential of EA‐TD‐SiQ to be used as a PDT agent in endometrial cancer. [ABSTRACT FROM AUTHOR]

BACKGROUND: The first evidence of blood biochemical and proteomic changes in slow‐growing broiler chicken subjected to ritual slaughter like halal (HS) and jhatka (JS) without stunning and commercial slaughter with electrical stunning (ES) was decoded. RESULTS: Significant stress indicators like cortisol and triiodothyronine were markedly elevated in JS birds, whereas increased (P < 0.05) levels of lactate dehydrogenase and creatine kinase were observed in JS and ES birds. Two‐dimensional gel electrophoresis coupled to MALDI‐TOF MS elucidated the overabundance of glyceraldehyde‐3‐phosphate dehydrogenase and l‐lactate dehydrogenase that are positively correlated with stress in JS broilers. Bioinformatic analysis explored the multifaceted landscape of molecular functions. CONCLUSION: The study has uncovered that ritual slaughter performed without stunning against commercial slaughter with ES practices elicit varying levels of stress as evident from blood biochemistry and novel protein markers. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

In this research, the differences in protein and fatty acid composition among camel, cow and goat milks were analyzed by gel electrophoresis (GE) and gas chromatography (GC). The GE results showed that there were differences in the proportion of proteins in camel milk from different regions, and the production process had a significant effect on both camel milk protein and milk fat globule membrane protein composition. There were significant differences in protein composition among camel, cow and goat milks, and camel milk did not contain β-lacglobulin or αs1-casein, while cow and goat milks did not contain immunoglobulin G2 (IgG2) or IgG3. Therefore, GE could be used for the identification of camel milk mixed with cow or goat milk. The GC results showed that 31 fatty acids were detected in both camel and goat milks, and 29 fatty acids in cow milk. Camel milk contained fewer short-chain fatty acids than cow and goat milks, while the contents of most long-chain fatty acids were significantly higher than those in cow and goat milks. Principal component analysis (PCA) of the fatty acid composition data showed a clear separation of milks from different species and a clear discrimination between pure and adulterated camel milk. PCA can be used for the identification and analysis of fatty acids in camel milk mixed with cow milk and goat milk.

During the selection of crops aimed at creating high-fiber varieties, it is important to determine the patterns of fiber formation at the molecular level and the genotype specificity of cellulose synthesis processes, to study the efficiency of the genes responsible for cellulose synthesis at various stages of plant ontogenesis. The aim of the work was to study the expression of flax cellulose synthase genes. The objects of the study were 8 samples of flax (genus Linum), differing in fiber content: 7 cultivarеs of Linum usitatissimum L., of which 5 cultivarеs of fiber flax, 2 cultivarеs of oilseed flax, as well as one sample of large-flowered flax (L. grandiflorum Desf.). The studied cultivar were grown in a growth chamber. Samples of plant material were taken on the 10th, 20th, 30th, 40th and 50th day after germination. RNA extraction and cDNA synthesis were carried out. RNA sequences of CesA1, CesA4, CesA6, CesA7 were obtained and gene-specific primers for PCR were developed. Gene expression was determined by semi-quantitative RT-PCR, and reaction products were separated by size using a 1.2% agarose gel. Based on the data obtained, the presence of phase- and genotype-specificity in the process of expression of cellulose synthases of the CesA family in the flax stem was established. Cellulose synthases of the primary cell wall of flax (CesA1, CesA6) show a dynamic increase in expression up to the «herringbone» stage, and then the expression gradually decreases towards the onset of rapid growth phase, while cellulose synthases of the secondary cell wall (CESA4, CESA7) are characterized by a constant increase in expression from the first days of the development of seedlings to the phase of rapid growth.

Background: Colon cancer is one of the most prevalent tumors in the digestive tract, and its stemness feature significantly contribute to chemoresistance, promote the epithelial-mesenchymal transition (EMT) process, and ultimately lead to tumor metastasis. Therefore, it is imperative for researchers to elucidate the molecular mechanisms underlying the enhancement of stemness feature, chemoresistance, and EMT in colon cancer. Methods: Sphere-formation and western blotting assays were conducted to assess the stemness feature. Edu, flow cytometry, and cell viability assays were employed to evaluate the chemoresistance. Immunofluorescence and western blotting assays were utilized to detect EMT. Immunoprecipitation, ubiquitination, agarose gel electrophoresis, chromatin immunoprecipitation followed by quantitative PCR (chip-qPCR), and dual luciferase reporter gene assays were employed for mechanistic investigations. Results: We demonstrated a markedly higher expression level of OTUB2 in colon cancer tissues compared to adjacent tissues. Furthermore, elevated OTUB2 expression was closely associated with poor prognosis and distant tumor metastasis. Functional experiments revealed that knockdown of OTUB2 attenuated stemness feature of colon cancer, enhanced its sensitivity to oxaliplatin, inhibited its EMT process, ultimately reduced the ability of tumor metastasis. Conversely, overexpression of OTUB2 exerted opposite effects. Mechanistically, we identified OTUB2 as a deubiquitinase for SP1 protein which bound specifically to SP1 protein, thereby inhibiting K48 ubiquitination of SP1 protein. The SP1 protein functioned as a transcription factor for the GINS1, exerting its regulatory effect by binding to the 1822–1830 region of the GINS1 promoter and enhancing its transcriptional activity. Ultimately, alterations in GINS1 expression directly regulated stemness feature, chemosensitivity, and EMT progression in colon cancer. Conclusion: Collectively, the OTUB2/SP1/GINS1 axis played a pivotal role in driving stemness feature, chemoresistance, and EMT in colon cancer. These results shed new light on understanding chemoresistance and metastasis mechanisms involved in colon cancer. [ABSTRACT FROM AUTHOR]

Organic molecule‐mediated noncanonical DNA self‐assembly expands the standard DNA base‐pairing alphabets. However, only a very limited number of small molecules have been recognized as mediators because of the tedious and complicated experiments like crystallization and microscopy imaging. Here we present an integrative screening protocol incorporating molecular dynamics (MD) for fast theoretical simulation and native polyacrylamide gel electrophoresis for convenient experimental validation. Melamine, the molecule that was confirmed mediating noncanonical DNA base‐pairing, and 38 other candidate molecules were applied to demonstrate the feasibility of this protocol. We successfully identified seven stable noncanonical DNA duplex structures, and another eight novel structures with sub‐stability. In addition, we discovered that hairpins at both ends can significantly stabilize the noncanonical DNA structures, providing a guideline to design small organic molecule‐incorporated DNA structures. Such an efficient screening protocol will accelerate the design of alternative DNA‐molecule architectures beyond Watson–Crick pairs. Considering the wide range of potential mediators, it will also facilitate applications such as noncovalent, highly dense loading of drug molecules in DNA‐based delivery system and probe design for sensitive detection of certain molecules. [ABSTRACT FROM AUTHOR]

Copyright of Shipin Kexue/ Food Science is the property of Food Science Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

We investigated suitable culture conditions for the production of the blue pigment phycocyanin (PC) from the unique filamentous cyanobacteria Pseudanabaena sp. ABRG5-3 and Limnothrix sp. SK1-2-1. White, green, or red LED irradiation at 30 μmol photons/m2/s was effective for phycocyanin production when compared with Arthrospira platensis (Spirulina) sp. NIES-39, which is generally grown under high light irradiation. To investigate the safety of the cyanobacteria, ABRG5-3 cells were subjected to Ames (reverse mutation) tests and single oral-dose rat studies, which revealed non-mutagenic and non-toxic properties. When three purified phycocyanins (abPC, skPC, and spPC) were subjected to agarose gel electrophoresis, they showed different mobility, indicating that each phycocyanin has unique properties. abPC exhibited strong antiglycation activities as novel function. • We assessed cultivation conditions and phycocyanin production of 3 cyanobacteria. • We tested various light irradiation conditions in terms of quality and intensity. • We found that ABRG5-3 cells are non-mutagenetic and non-toxic. • Each phycocyanin has unique properties depending on bacterial species. • The phycocyanins showed strong antiglycation activity. [ABSTRACT FROM AUTHOR]

Bird sex determination is fundamental in various ecological and biological studies, although many avian species cannot be sexed visually due to their monomorphic and/or monochromatic appearance. Thus, reliable laboratory methods for sexing are a prerequisite. Most avian nestlings lack sex-related signs, including the Eurasian pygmy owl (Glaucidium passerinum). We performed laboratory sex determination analysis of this species using blood samples of 242 juveniles and nine adults. It relied on the qPCR of the specific intron from the chromo-helicase DNA-binding protein 1 gene. We tested three primer sets, the P2/P8, 2550F/2718R, and CHD1F/CHD1R, commonly used for bird laboratory sexing. The outcomes were displayed on an agarose gel electrophoresis and a plot from melt curve analysis, which had not been previously conducted in Eurasian pygmy owls. We found that only primer set CHD1F/CHD1R proved reliable, as the only one determined sex with one and two band/s and peak/s on the electrophoresis and the melt curve plot for males and females, respectively. The other two primer pairs failed and depicted one band/peak in all specimens regardless of their sex. Therefore, we recommend performing Eurasian pygmy owls' laboratory sexing by qPCR with CHD1F/CHD1R primers only. [ABSTRACT FROM AUTHOR]

Background: Colorectal cancer (CRC) remains a global health concern with persistently high incidence and mortality rates. However, the specific pathogenesis of CRC remains poorly understood. This study aims to investigate the role and pathogenesis of serine and arginine rich splicing factor 10 (SRSF10) in colorectal cancer. Methods: Bioinformatics analysis was employed to predict SRSF10 gene expression in CRC patients. Functional experiments involving SRSF10 knockdown and overexpression were conducted using CCK8, transwell, scratch assay, and flow cytometry. Additionally, the PRIdictor website was utilized to predict the SRSF10 interaction site with RFC5. The identification of different transcripts of SRSF10-acting RFC5 pre-mRNA was achieved through agarose gel electrophoresis. Result: The knockdown of SRSF10 inhibited the proliferation and migration ability of CRC cells, while promoting apoptosis and altering the DNA replication of CRC cells. Conversely, when SRSF10 was highly expressed, it enhanced the proliferation and migration ability of CRC cells and caused changes in the cell cycle of colorectal cancer cells. This study revealed a change in the replicating factor C subunit 5 (RFC5) gene in colorectal cancer cells following SRSF10 knockdown. Furthermore, it was confirmed that SRSF10 increased RFC5 exon2-AS1(S) transcription variants, thereby promoting the development of colorectal cancer through AS1 exclusion to exon 2 of RFC5. Conclusion: In summary, this study demonstrates that SRSF10 promotes the progression of colorectal cancer by generating an aberrantly spliced exclusion isoform of AS1 within RFC5 exon 2. These findings suggest that SRSF10 could serve as a crucial target for the clinical diagnosis and treatment of CRC. [ABSTRACT FROM AUTHOR]

The ability to recognize and differentiate between conspecifics and heterospecifics as well as their signals is critical for the coexistence of closely related species. In the genus Rattus , species are morphologically similar and multiple species often coexist. Here, we investigated the interspecific recognition and signal differentiation of two sympatric rat species, the brown rat (Rattus norvegicus , RN) and the Asian house rat (Rattus tanezumi , RT). In a two-way choice test, both RN and RT females showed a preference for conspecific male rats to heterospecific ones. RT females showed a significant preference for accessible urine of males of same species to those of other species, but not for the inaccessible urine. On the other hand, there were significant differences in the structural characteristics of the ultrasonic vocalization emitted by males of these two rat species. Sodium dodecyl sulphate‒polyacrylamide gel electrophoresis (SDS‒PAGE) and isoelectric focusing electrophoresis unveiled that major urinary proteins (MUPs) in voided urine were more highly expressed in RN males versus RT males. The interspecific differences of urinary volatile compounds were also discussed. In conclusion, female rats had the ability to distinguish between males of either species. [ABSTRACT FROM AUTHOR]

1. Epidemiological surveillance of Salmonella spp. serves as a primary tool for maintaining the health of poultry flocks. Characterising circulating serotypes is crucial for implementing control and prevention measures. This study conducted phenotypic and molecular characterisation of S. enterica Pullorum, S. enterica Heidelberg, and S. enterica Corvalis isolated from broiler chickens during slaughtering. 2. All strains were susceptible to gentamicin, neomycin and norfloxacin. However, resistance rates exceeded 50% for ciprofloxacin and tiamulin, irrespective of the serotype. Approximately 64% of strains were classified as multidrug-resistant, with S. enterica Heidelberg strains exhibiting significantly higher overall resistance. The isolates demonstrated the ability to adhere and produce biofilm at a minimum of three temperatures, with S. enterica Pullorum capable of biofilm production at all temperatures encountered during poultry rearing. 3. Each strain possessed between two and seven different virulence-associated genes. Genetic similarity, as indicated by pulsed field gel electrophoresis, exceeded 90% for all three serotypes and strains were classified in the R5 ribotype by PCR, regardless of serotype. Sequencing revealed high similarity among all strains, with homology ranging from 99.61 to 100% and all were classified to a single cluster. 4. The results suggested a clonal relationship among the strains, indicating the possible circulation of a unique clonal group of S. enterica Pullorum in the southern region of Brazil. [ABSTRACT FROM AUTHOR]