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Membrane proteins are underrepresented during proteome characterizations, primarily owing to their lower solubility. Sodium dodecyl sulfate (SDS) is favored to enhance protein solubility but interferes with downstream analysis by mass spectrometry. Here, we present an improved workflow for SDS depletion using transmembrane electrophoresis (TME) while retaining a higher recovery of membrane proteins. Though higher levels of organic solvent lower proteome solubility, we found that the inclusion of 40% methanol provided optimal solubility of membrane proteins, with 86% recovery relative to extraction with SDS. Incorporating 40% methanol during the electrophoretic depletion of SDS by TME also maximized membrane protein recovery. We further report that methanol accelerates the rate of detergent removal, allowing TME to deplete SDS below 100 ppm in under 3 min. This is attributed to a three-fold elevation in the critical micelle concentration (CMC) of SDS in the presence of methanol, combined with a reduction in the SDS to protein binding ratio in methanol (0.3 g SDS/g protein). MS analysis of membrane proteins isolated from the methanol-assisted workflow revealed enhanced proteome detection, particularly for proteins whose pI contributed a minimal net charge and therefore possessed reduced solubility in a purely aqueous solvent. This protocol presents a robust approach for the preparation of membrane proteins by maximizing their solubility in MS-compatible solvents, offering a tool to advance membrane proteome characterization.

This work presents the results of research on submicro- and nanocomposites with gradient properties, produced in a planned electrophoretic process. Epoxy-resin-based samples were filled with TiO2 particles of three different sizes (13 nm, 38 nm, and Eav (0.0 Vmm−1, 125 Vmm−1, 250 Vmm−1, and 500 Vmm−1) for 1 h each. Changes in selected dielectric parameters (dielectric constant εr and dielectric loss factor tanδ) of the composites were analyzed using broadband dielectric spectroscopy (10−1 Hz to 105 Hz). The influence of the Eav and the current i(t) flowing through the sample material and the Joule heat generated in it on the resin curing process and the final gradient of dielectric parameters were investigated. The results show that the degree of modification of the εr gradient increases with increasing Eav and is more pronounced in the case of TiO2 nanoparticles. The largest modifications in the εr and tanδ were obtained for nanoparticles with a diameter of 13 nm at Eav = 500 Vmm−1, while the lowest for particles < 1 µm at Eav = 125 Vmm−1. The effect of electrophoresis on the dielectric parameters is significant, especially near the anode region. Increasing the concentration of TiO2 particles at the anode occurs at the expense of reducing their concentration in the remaining volume of the sample. The test results clearly demonstrate the importance of particle dimension and electric field strength for the gradient modification of the properties of the epoxy composite using electrophoresis. Numerical simulations of electric field stresses in the epoxy resin during the electrophoresis process, performed in the COMSOL program, revealed a significant increase in the E field strength in the areas close to the anode and cathode.

Bimodal optical-electrical data generated when a 20 nm diameter silica (SiO2) nanoparticle was trapped by a plasmonic nanopore sensor were simulated using Multiphysics COMSOL and compared with sensor measurements for closely matching experimental parameters. The nanosensor, employed self-induced back action (SIBA) to optically trap nanoparticles in the center of a double nanohole (DNH) structure on top a solid-state nanopores (ssNP). This SIBA actuated nanopore electrophoresis (SANE) sensor enables simultaneous capture of optical and electrical data generated by several underlying forces acting on the trapped SiO2 nanoparticle: plasmonic optical trapping, electroosmosis, electrophoresis, viscous drag, and heat conduction forces. The Multiphysics simulations enabled dissecting the relative contributions of those forces acting on the nanoparticle as a function of its location above and through the sensor's ssNP. Comparisons between simulations and experiments demonstrated qualitative similarities in the optical and electrical time-series data generated as the nanoparticle entered and exited from the SANE sensor. These experimental parameter-matched simulations indicated that the competition between optical and electrical forces shifted the trapping equilibrium position close to the top opening of the ssNP, relative to the optical trapping force maximum that was located several nm above. The experimentally estimated minimum for the optical force needed to trap a SiO2 nanoparticle was consistent with corresponding simulation predictions of optical-electrical force balance. The comparison of Multiphysics simulations with experiments improves our understanding of the interplay between optical and electrical forces as a function of nanoparticle position across this plasmonic nanopore sensor.

Abstractr: In order to improve the weak binding force and easy pull-out of the traditional carbon fiber (CF) and concrete interface, this paper prepared three-dimensional multi-scale carbon nanotube-fiber (MCNF) by grafting carbon nanotubes on the surface of carbon fibers by electrophoresis, and carried out computer simulations. The results showed that compared with ordinary carbon fibers, the attraction between the MCNF fibers surface and the concrete interface is stronger, and it is not easy to pull out. At the same time, Plain concrete (PC), CF concrete (CF content as 0.1% of concrete volume) and MCNF concrete (MCNF content as 0.1% of concrete volume) were prepared, and dynamic mechanical tests were carried out by using a Φ100 mm Hopkinson pressure bar test (SHPB) experimental device., It is found that compared with carbon fiber, the interface between MCNF and concrete is more tightly bonded and difficult to pull out, so it has better effect on improving the mechanical properties of concrete. Finally, MCNF enhencement effect was introduced to simplify and modify the Zhu-Wang-Tang (ZWT) constitutive model which is the damage model of traditional and the simplified model conforms to the actual situation of MCNF concrete. The theoretical curve based on the constitutive model is also similar to the actual test curve, proving the correctness of the constitutive model.The research results provide references for improving the impact load resistance of concrete in practical engineering.

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Background: Although the global thalassemia zone covers Bangladesh, there are very limited studies conducted in this region. Therefore, the focus of our study is to understand the prevalence and burden of thalassemia and hemoglobinopathy in Bangladesh. Methods: The analysis was based on a retrospective evaluation of laboratory diagnoses between 2007 January and 2021 October. A total of 8503 specimens were sampled and analyzed which were either referred by corresponding physicians or self-referred. This was neither any epidemiological nationwide survey nor was the study population chosen randomly. Hematological data were obtained through capillary zone electrophoresis and corresponding complete blood count. Results: 1971 samples (~23.18% of the total) were found with at least one inherited hemoglobin disorder. The most common hemoglobin disorder observed was the hemoglobin E (Hb E) trait (10.67%), followed by the β-thalassemia trait (8.4%), homozygotic Hb E (1.59%), and Hb E/β-thalassemia (1.58%). Other variants found in this study with minimal percentages were Hb N-Seattle, Hb S, Hb D-Punjab, Hb Lepore, Hb C, Hb Hope, Hb H, and hereditary persistence of fetal hemoglobin. Discussion: The pattern of thalassemia and hemoglobinopathy in our study is diverse and heterogeneous. A broad and detailed spectrum of such inherited hemoglobin disorders will ultimately be helpful in implementing nationwide thalassemia management and strategy policy in Bangladesh. [ABSTRACT FROM AUTHOR]

Based on the analysis of patients with Peutz-Jeghers syndrome (PJS), Serine threonine kinase11 (STK11) is known as a tumor suppressor gene, which is involved in cell polarization, regulation of apoptosis, and DNA damage response. In this case report study, we examined STK11 gene sequencing in a 42-year-old woman with mucocuta neous pigmentation and positive family history. Endoscopy and colonoscopy showed >1000 polyps throughout the stomach/colon (PJ-type hamartomas). The larger polyp in the stomach was resected and the small bowel imaging detected multiple jejunum/ileum small polyps. The data released from the sequencing results revealed five alterations in exons 1 to 5. The major mutation in stop codon was reported as converted to the amino acid tryptophan (TRP) to tyrosine (TER). The TGG codon was converted to TAG by mutation. Finally, another novel mutation in STK11 stop codon as a 'de novo' variant was seen. It is predicted that stop codon mutations make the affected person susceptible to developing colorectal cancer. [ABSTRACT FROM AUTHOR]

Cataphoresis varnishing enables an organic coating to form on an aluminum substrate, thus increasing its corrosion resistance and durability. Cataphoresis varnishing is known to ensure a high adhesion of the created cataphoresis layer and a good homogeneity of this layer, even on surfaces with complex geometry. This paper aimed to optimize the deposition process and to analyze and evaluate the thickness of a cataphoresis layer formed on an aluminum substrate from AW 1050—H24 material. In total, 30 separate samples were created in accordance with the Design of Experiments methodology, using a central composite plan. The independent input factors in the study were: the electrical voltage (U) and deposition time in the cataphoresis varnishing process (tKTL) at the polymerization times of 15 min, 20 min, and 25 min, respectively. The results of the statistical analysis showed that the voltage accounted for 33.82% of the change in the thickness of the created layer and the deposition time contributed 28.67% to thi change. At the same time, the interaction of the voltage and deposition time (p < 0.0001) accounted for 20.25% of the change in the thickness of the layer under formation. The regression model that was constructed showed a high degree of prediction accuracy (85.8775%) and its use as a function for nonlinear optimization provided a maximum layer thickness th of max = 26.114 µm, at U = 240 V and tKTL = 6.0 min, as was proven under experimental conditions. [ABSTRACT FROM AUTHOR]

A novel electrophoretic separation technique is presented, where continuous electrophoretic separation is demonstrated using free flowing liquid foams. Continuous foam electrophoresis combines the principle of capillary electrophoresis and interactions between analytes and the electrical double layer, with the ability of Free Flow Electrophoresis to continuously separate and recover analytes automatically. A liquid foam is used to provide a network of deformable micro and nano channels with a high surface area, presenting a novel platform for electrophoresis, where interfacial phenomena could be exploited to modify analyte migration. The main purpose of this paper is to present a proof-of-concept study and provide fundamental understanding of a complex foam system in continuous separation mode, i.e., flowing liquid foam under an external electric field with electrophoresis and chemical reactions at the electrodes continuously changing the system. Liquid foam is generated using a mixture of anionic and non-ionic surfactants and pumped through a microfluidic separation chamber between two electrodes. The effectiveness of the device is demonstrated using a dye mixture containing a neutral dye and an anionic dye. At the outlet, the foam is separated and collected into five fractions which are individually probed for the concentration of the two dyes used. The anionic dye was concentrated up to 1.75 (±0.05) times the initial concentration in a select outlet, while the neutral dye concentration remained unchanged in all outlets, demonstrating the potential for electrophoretic foam separations. [ABSTRACT FROM AUTHOR]

The goal of this paper is to assess the mutagenic and genotoxic potentials of avocado oil made from the fruit pulp of Persea Americana, a member of the Lauraceae family. Michigan Cancer Foundation-7 (MCF-7) cells are used in the 3-4,5 dimethylthiazol-2yl-2,5-diphenyl tetrazolium bromide (MTT) test to examine the possible antiproliferative and cytostatic qualities of different doses of avocado oil, and MCF-7 cells are used in the comet assay to examine the potential cytostatic effects of avocado oil extracted from the avocado fruit. DNA in human breast cancer cells is partially damaged by avocado oil. However, DNA damage at low, medium, and high levels was discovered in the positive control. Without positive control, the DNA damage level falls in the low, middle, and high ranges. The MTT assay shows that avocado oil exerts a dose-dependent cytostatic impact on human breast cancer MCF-7 cells with an IC50 value of 379.2 µg/mL, which is the IC50 value that causes genotoxicity in the comet assay. [ABSTRACT FROM AUTHOR]

The tailored physical properties of TiO2 are of significant importance in various fields and, as such, numerous methods for modifying these properties have been introduced. In this study, we present a novel method for doping Fe into TiO2 via the anodic dissolution of iron. The optimal conditions were determined to be an application of 200 V to acetylacetone (acac)/EtOH medium for 10 min, followed by the addition of TiO2 to the solution, sonication for 30 min, stirring at 80 °C, and drying. The resulting powder was calcined at 400 °C for 3 h, and characterization was conducted using XRD, FTIR, TEM, and UV-vis. The synthesized powder revealed the successful doping of Fe into the TiO2 structure, resulting in a decrease in the optical band gap from 3.22 to 2.92 eV. The Fe-TiO2 was then deposited on a metal substrate via the electrophoretic (EPD) technique, and the weight of the deposited layer was measured as a function of the applied voltage and exposure time. FESEM images and EDX analysis confirmed that the deposited layer was nanostructured, with Fe evenly distributed throughout the structure. [ABSTRACT FROM AUTHOR]

The texture of surimi-like gels made from the protein isolate extracted from catfish byproducts has been proven to be brittle and lack elasticity. To address this issue, varying levels of microbial transglutaminase (MTGase) from 0.1 to 0.6 units/g were applied. MTGase had little effect on the color profile of gels. When MTGase at 0.5 units/g was employed, hardness, cohesiveness, springiness, chewiness, resilience, fracturablity, and deformation were increased by 218, 55, 12, 451, 115, 446, and 71%, respectively. A further increase in added MTGase did not lead to any textural improvement. In comparison to the gels made from fillet mince, the gels made from protein isolate were still lower in cohesiveness. Due to the activated endogenous transglutaminase, a setting step enhanced the textural properties of gels made from fillet mince. However, because of the endogenous proteases-induced protein degradation, the setting step led to a texture deterioration of the gels made from protein isolate. Gels made from protein isolate showed 23–55% higher solubility in reducing solution than in non-reducing solution, suggesting the vital role of disulfide bonds in the gelation process. Due to the different protein composition and conformation, fillet mince and protein isolate exhibited distinct rheological properties. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed the highly denatured protein isolate was susceptible to proteolysis and prone to disulfide formation during the gelation process. It also revealed that MTGase had an inhibitory effect on the proteolysis induced by endogenous enzymes. In view of the susceptibility of the protein isolate to proteolysis during gelation, future research should consider including other enzyme inhibitory agents in the presence of MTGase to improve the gel texture. [ABSTRACT FROM AUTHOR]

The purpose of this study was to investigate the improvement in techno-functional properties of mung bean protein isolate (MBPI) treated with microbial transglutaminase (MTG), including water- and oil-holding capacity, gelling properties, and emulsifying capacity. MBPI dispersions were incubated with MTG (5 U/g of protein substrate) at 45 °C with constant stirring for 4 h (MTM4) or 8 h (MTM8). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that MTG treatment for different durations increased the amount of high-molecular-weight proteins in MBPI, and most of the cross-linking by MTG was terminated at 8 h. Improved water-holding capacity, gelling properties, emulsifying capacity, and stability were observed after MTG treatment, and decreased protein solubility and surface hydrophobicity were observed. Furthermore, the texture of the heat-induced gels made from MTG-treated MBPI was evaluated using a texture analyzer. MTG treatment increased the hardness, gumminess, chewiness, and adhesiveness of the heat-induced gels. Field-emission scanning electron microscopy demonstrated the enhanced hardness of the gels. This research reveals that MTG-catalyzed cross-linking may adjust the techno-functional properties of MBPI, allowing it to be used as a soy protein alternative in food products, such as plant-based and processed meats. [ABSTRACT FROM AUTHOR]

Background: The activity and purity of Fibrin Sealant Grifols are described in this article. This fibrin sealant contains two components (fibrinogen and thrombin) that mimic the final stages of clotting. Fibrin sealants are primarily used as an adjunct to hemostasis for mild to moderate surgical bleeding in adults when standard surgical techniques are ineffective or impractical. Objective: The aim of these studies was to assess the biochemical and functional characteristics of Fibrin Sealant Grifols. Methods: Total protein, plasma protein impurities, and molecular forms were analyzed for both components. Clottable fibrinogen and thrombin activity were also measured. Both components were analyzed by SDS-PAGE and Western blot. The fibrin sealant clot was functionally characterized. Results: The components had minimal contamination by other plasma proteins and no aggregates or degradation products. The fibrinogen component was almost completely clottable fibrinogen. The thrombin component contains albumin. The fibrinogen component formed a fibrin polymer and clots with tensile strength similar to literature values. The application device applied the components in a 1:1 volume ratio. Clotting time was ∼ 5 s when applied with the drip tip and nearly instantaneous when applied with the airless spray apparatus. Conclusion: This fibrin sealant was shown to contain highly purified fibrinogen and thrombin delivered in a 1:1 volume ratio. These properties contribute to the clinical safety and efficacy of this product for many applications. [ABSTRACT FROM AUTHOR]

Diffusiophoresis of a weakly charged liquid metal droplet (LMD) is investigated theoretically, motivated by its potential application in drug delivery. A general analytical formula valid for weakly charged condition is adopted to explore the droplet phoretic behavior. We determined that a liquid metal droplet, which is a special category of the conducting droplet in general, always moves up along the chemical gradient in sole chemiphoresis, contrary to a dielectric droplet where the droplet tends to move down the chemical gradient most of the time. This suggests a therapeutic nanomedicine such as a gallium LMD is inherently superior to a corresponding dielectric liposome droplet in drug delivery in terms of self-guiding to its desired destination. The droplet moving direction can still be manipulated via the polarity dependence; however, there should be an induced diffusion potential present in the electrolyte solution under consideration, which spontaneously generates an extra electrophoresis component. Moreover, the smaller the conducting liquid metal droplet is, the faster it moves in general, which means a smaller LMD nanomedicine is preferred. These findings demonstrate the superior features of an LMD nanomedicine in drug delivery. [ABSTRACT FROM AUTHOR]

Objective Isolated low hemoglobin A2 (HbA2) is rarely encountered in our clinical practice using capillary zone electrophoresis. The study goal was to characterize the work-up at our institution of patients with low HbA2. Methods Patients with low HbA2 and a control cohort with normal capillary zone electrophoresis were identified and relevant information extracted from the medical record. Results Of 44 patients with isolated decreased HbA2, 28 (64%) had corresponding complete blood count/ferritin values. Compared to control patients, patients with low HbA2 were more likely to have iron deficiency and demonstrated a more microcytic, hypochromic blood picture. However, 46% (13/28) of patients with low HbA2 and ferritin for evaluation did not have iron deficiency. Only 2 patients had genetic testing. Conclusion This study redemonstrates the association between low HbA2 and iron deficiency and reinforces the need for iron indices to interpret capillary zone electrophoresis results. Our study population showed incomplete or absent iron studies in most cases. [ABSTRACT FROM AUTHOR]

Objective Our aim was to establish a chemiluminescence method for detecting anti-transmembrane protein (p7) antibody in the serum of patients with hepatitis C virus (HCV) infection. Methods The p7 gene was amplified by polymerase chain reaction using the plasmid PUC-p7 containing the p7 nucleic acid sequence of the HCV 1b genotype as the template, and recombinant plasmid pGEX-KG-p7 was constructed. After p7 fusion, the protein was induced and expressed in the prokaryote, extracted, and purified; the anti-p7 antibody detection kit was prepared, and its efficacy was evaluated. Results The plasmid pGEX-KG-p7 was constructed correctly, and p7 fusion protein was obtained. The methodological indexes of the kit, the precision test, blank limit and detection limit, etc, met the requirements. The positive rate of serum anti-p7 antibody in 45 patients with HCV infection was 20%. Conclusions The kit can be used in screening diagnosis, condition monitoring, prognosis, and disease mechanism and epidemiological study of HCV infection. The p7 protein has immune response in HCV-infected patients. [ABSTRACT FROM AUTHOR]

Background and Aim: Oral infections are common among people of any age and can trigger systemic inflammation. The microbiota is a diverse group of microorganisms that play important roles in metabolism, immune function, and homeostasis. Oral microbiota in human atherosclerotic plaques has been identified using various techniques. Therefore, the focus of this study was to determine the correlation between metabolic parameters and oral microbiota composition in patients with atherosclerosis using Denaturing Gradient Gel Electrophoresis (DGGE) assays. Materials and Methods: In this case-control study, saliva samples were collected from 139 patients with atherosclerosis and healthy individuals from Imam Ali Cardiovascular Hospital, Kermanshah, Iran. After DNA extraction, PCR products were examined and evaluated using DGGE assays. Results: The study included 89 (36%) patients with a history of atherosclerosis and 50 (36%) healthy individuals. There was a significant relationship between the mean total cholesterol, Low-Density Lipoprotein (LDL), Fasting Blood Sugar (FBS), and Blood Urea Nitrogen (BUN) in the two groups. However, there was no significant difference in the mean high-density lipoprotein (HDL) and Triglyceride levels between the study groups. Conclusion: Our results showed a relationship between metabolic parameters and oral microbiota composition in patients with atherosclerosis. Additionally, our results indicated that the DGGE assay is a useful method for diagnosing and comparing the oral microbiota of people with atherosclerosis and healthy individuals. Therefore, further examination of the oral microbiota is necessary to determine its potential as a biomarker for atherosclerosis. [ABSTRACT FROM AUTHOR]

Background and Aim: Entamoeba histolytica and Giardia intestinalis are two common parasites associated with gastrointestinal infections and diarrheal diseases worldwide. This study investigated the prevalence, molecular characterization, and phylogenetic relationships of E. histolytica and G. intestinalis among patients with diarrhea and intestinal disorders in hospitals across Iraq. Materials and Methods: Stool samples were collected from individuals presenting with gastrointestinal symptoms, and a microscopic examination was performed to screen for the presence of E. histolytica and G. intestinalis. DNA was extracted from positive samples, and PCR amplification targeting the 16S rRNA gene for E. histolytica and the triose phosphate isomerase (TPI) gene for G. intestinalis was carried out. The resulting PCR products were subjected to agarose gel electrophoresis, purified, and sequenced. Phylogenetic trees were constructed using the Maximum Likelihood method based on the Tamura-Nei model, and bootstrap analysis was performed. Results: Among the studied samples, 27.8% (n=85) were positive for E. histolytica, while 19.6% (n=60) were positive for G. intestinalis. Sequence analysis confirmed the presence of E. histolytica and G. intestinalis by matching the obtained sequences with those available in the NCBI gene bank. The phylogenetic analysis revealed the genetic diversity and relatedness of the E. histolytica and G. intestinalis isolates. Clustering patterns specific to the Iraqi population were observed, suggesting potential regional transmission dynamics. Conclusion: This study provides valuable insights into the prevalence, molecular characterization, and phylogenetic relationships of E. histolytica and G. intestinalis in Iraqi patients with diarrhea and intestinal disorders. [ABSTRACT FROM AUTHOR]

Simple Summary: Waldenström's macroglobulinemia is a lymphoproliferative disorder rarely described in veterinary medicine. According to the human classification of hematopoietic tumors, this disease is defined as a subtype of lymphoplasmacytic lymphoma with bone marrow involvement and monoclonal gammopathy of IgM type. Cytopathology is a convenient and accurate diagnostic procedure to diagnose lymphoma in dogs. However, ancillary techniques, such as PCR for antigen receptor rearrangement, flow cytometry, immunohistochemistry, and histology, are usually required to fully characterize the grade and phenotype, especially in ambiguous cases. Immunosecretory lymphoproliferative disorders result from malignant clonal expansion of B-cells and are usually characterized by hyperglobulinemia, although normoglobulinemia can also occur, hypoglobulinemia being rarer. Monoclonal gammopathy can be suspected on serum protein electrophoresis as it usually migrates as a narrow peak in the γ or β fractions, but immunofixation is required for confirmation and characterization. This case report describes clinicopathologic features, diagnosis, treatment, and the outcome of Waldenström's macroglobulinemia in a 2-year-old neutered female Small Munsterlander dog with atypical bimorphic plasmacytoid differentiation and unusual M-protein migration on serum protein electrophoresis. A 2-year-old neutered female Small Munsterlander dog was presented for an insect bite. Physical examination revealed a poor body condition, a peripheral lymphadenomegaly, and suspected splenomegaly. A complete blood count (Sysmex XN-V) revealed marked leukocytosis with lymphocytosis and abnormal dot plots. An abnormal monomorphic lymphoid population and marked rouleaux formation were noted on the blood smear. Lymph node aspirates contained an atypical bimorphic population of lymphocytes, either with a plasmacytoid or a blastic appearance. This double population was also found in the spleen, liver, bone marrow, tonsils, and other tissues. Peripheral blood and lymph node clonality assays revealed clonal BCR gene rearrangement. Flow cytometry revealed a mixed population of small-sized B-cells (CD79a+ CD21+ MHCII+) and medium-sized B-cells (CD79a+ CD21− MHCII−) in lymph nodes and a dominant population of small-sized mature B-cells (CD21+ MHCII+) in peripheral blood. Though normoproteinemic, serum protein electrophoresis revealed an increased α2-globulin fraction with an atypical restricted peak, identified as monoclonal IgM by immunofixation. Urine protein immunofixation revealed a Bence-Jones proteinuria. A diagnosis of Waldenström's macroglobulinemia was made. Chemotherapy was initiated, but the dog was euthanized 12 months after the initial presentation due to marked clinical degradation. [ABSTRACT FROM AUTHOR]

Background: Parallel to the growth of the oral healthcare market, there is a constantly increasing demand for natural products as well. Many customers prefer products that contain fewer toxic agents, therefore providing an environmentally friendly solution with the benefit of smaller risk to the user. Medieval and early modern medicinal knowledge might be useful when looking for natural, herbal-based components to develop modern products. Along with these considerations we created, tested, and compared an entirely natural mouthwash, named Herba Dei. Methods: The manufacturing procedure was standardized, and the created tincture was evaluated by GC/MS analysis for active compounds, experimentally tested in cell-based cytotoxicity, salivary protein integrity, cell-free antioxidant activity, anti-bacterial and anti-viral assays, and compared with three market-leading mouthwashes. Results: Our tincture did not show significant damage in the cytotoxicity assays to keratinocyte and Vero E6 cells and did not disrupt the low molecular weight salivary proteins. Its radical scavenging capacity surpassed that of two tested, partly natural, and synthetic mouthwashes, while its antibacterial activity was comparable to the tested products, or higher in the bacterial aerobic respiratory assay. The active compounds responsible for the effects include naturally occurring phenylpropanoids, terpenes, and terpenoids. Our mouthwash proved to be effective in vitro in lowering the copy number of SARS-CoV-2 in circumstances mimicking the salivary environment. Conclusions: The developed product might be a useful tool to impede the transmission and spread of SARS-CoV-2 in interpersonal contact and aerosol-generating conditions. Our mouthwash can help reduce the oral bacterial flora and has an antioxidant activity that facilitates wound healing and prevents adverse effects of smoke in the oral cavity. [ABSTRACT FROM AUTHOR]

The objective of this study was to compare the electrophoretic pattern of serum proteins in newborn calves, lambs, goat kids and piglets in precolostral blood samples. Blood samples were collected within 30 min of birth, prior to ingestion of first colostrum, and the concentrations of total proteins and protein fractions were analyzed using electrophoresis on agarose gel. The size and shape of the protein fractions on the electrophoretograms differed among the investigated animal species. Significant differences were found in the total protein values and all the separated protein fractions, as well as albumin to globulin ratio (p < 0.01 and p < 0.001). The mean total protein concentration in piglets was lower compared to ruminants. Albumin concentrations were markedly higher, and the concentrations of α1-globulins were lower, in neonatal ruminants compared to piglets. The values of α2-globulins were higher in lambs and goat kids, and lower in calves and piglets. An opposite tendency was found in the values of β-globulins: a detectable amount of γ-globulins was recorded in all evaluated animal species. Presented results suggest marked species related differences in the shape and size of protein fraction among neonates of farm animal species, and the importance of the evaluation of electrophoretograms with regard to these findings. [ABSTRACT FROM AUTHOR]

Mutation breeding is a significant means of increasing breeding efficiency and accelerating breeding process. In present study, we explored a new method for mutations inducing in rice (Oryza sativa L.) by using direct current electrophoresis bath (DCEB). The results showed that 20 mM NaCl solution is the optimal buffer, and the mortality of rice seeds followed an upward trend with increasing voltage and processing time of DCEB. By exploring the mutagenic effects of γ-irradiation and DCEB on seed vigor and physiological damages, we found that the physiological damages induced by DCEB on seed vigor were significant compared with that by γ-irradiation. We screened two mutants with low filled grain percentage and one mutant with abnormal hull from the M2 generations. These three mutants were confirmed to be authentic mutants based on 48 SSR markers followed by the protocol NY/T 1433–2014. Whole-genome resequencing detected a total of 503 and 537 polymorphisms in the two mutants, respectively, and the DCEB mutagenesis induced mainly InDel variants, while the exon region of mutant genes occupied a large proportion, especially the SNP variants, which occupied about 20% of the mutation sites in the exon region. [ABSTRACT FROM AUTHOR]

In the context of solving the problem of obtaining high quality dairy products from livestock, the issue of determining the type of beta-casein (A1 and A2) in the protein fraction of milk becomes essential. Purpose - to analyse the use of ACRS-PCR methods for differentiation of A¹ and A² alleles of bovine beta-casein locus. Genotyping features were analysed using the artificially created restriction site polymerase chain reaction method utilising TaqI and Ddel restriction endonucleases. The electrophoretic distribution of DNA fragments in agarose gels of various concentrations was used to analyse restriction patterns. Based on the results of bioinformatic analysis of the nucleotide reference sequences of the experimental fragment of the beta-casein gene, it was found that the primer system for the ACRS-PCR DdeI method is characterised by higher parameters of flanking efficiency of the target DNA site compared to the ACRS-PCR TaqI system due to significantly greater effectiveness of hybridisation of oligonucleotides on the target DNA. Based on the results of laboratory tests of both methods, it is proposed to use an additional procedure for analysing the fluorescence intensity of individual elements of restriction patterns, which allows reducing the number of false genotyping that occurs in both cases (based on the results of using both methods) due to the appearance of non-specific amplification/restriction fragments within the size of target restrictions. The application of the ACRS-PCR DdeI method provides more differentiated patterns of the corresponding genotypes in agarose gel compared to the ACRS-PCR TaqI method, but leads to higher material costs for conducting research. These disadvantages of using primer systems for ACRS-PCR of the beta-casein locus determine the relevance of developing alternative methods for typing A¹ and A² alleles which include allele-specific PCR. The use of results is promising for solving the problems of genotyping cattle individuals of different breeds by A ¹ and A² alleles of the beta-casein locus [ABSTRACT FROM AUTHOR]

Introduction. In the Ukrainian Carpathians, a special Carpathian liquid milk (CLM) coagulant is used for the production of traditional soft cheeses such as brynza and budz. Since CLM coagulant has not been studied before, the aim of the work was to characterize its milk coagulant and proteolytic properties. Materials and methods. Milk coagulants were obtained from farms in Vorokhta, Carpathians. Milk-clotting activity was examined by the Soxhlet method. The content of proteins was determined using the Kjeldahl method. Homogeneous αS1-, β- and ϰ-casein substrates were separated by preparative electrophoresis. The concentration of the products of casein substrates proteolysis was determined spectrophotometrically. The specificity of casein fractions proteolysis was established by analytical electrophoresis. Results and discussion. CLM coagulant has high milk coagulant activity, which is slightly lower than of the standard rennet enzyme and is close to the solid milk coagulant preparation Glek. It was found that the activity of CLM coagulant increases during its storage, which is apparently achieved through the proteolytic enzyme extraction from calf's stomach cells. The milk clotting time for fresh CLM coagulant is 159 min, and after 18 months of storage it is reduced to 49 min. Milk-clotting activity of the fresh CLM coagulant is 2515 Soxhlet Units (SU), whereas after storage during 18 months the milk-clotting activity increased to 8100 SU. In the production of Carpathian brynza and budz, it is recommended to use the amount of CLM coagulant, which provides the beginning of milk coagulation at 33-35 °C after 20-25 min. The analysis of the casein substrates proteolysis products by electrophoretic and spectrophotometric methods shows the high specificity of CLM coagulant to ϰ-casein. Such specificity is characteristic for the purified natural milk coagulant enzyme - chymosin. Along with that, CLM coagulant, unlike pepsin and, to a lesser extent, standard rennet, practically does not cleave αS1- and β-casein substrates. Conclusions. Milk-clotting activity of CLM coagulant during its storage for 18 months increases 3 times. According to the specificity of αS1-, β- and ϰ-casein substrates proteolysis, CLM coagulant is close to chymosin. [ABSTRACT FROM AUTHOR]

We describe a workflow for efficient, environmentally attentive, and sustainable practices related to routine agarose gel electrophoresis. The methods reduce plastic waste and improve efficiency, especially for the exhaustive screening of difficult-to-obtain plasmids. Sustainability is increased when agarose is used ten times over by virtue of a thorough recycling regimen. The workflow optimizes workspaces and standardizes lab practices for handling potentially hazardous waste, minimizing environmental harm. Safety, efficiency, and sustainability improve laboratory productivity, help minimize environmental contamination, and increase cost-effectiveness. [ABSTRACT FROM AUTHOR]

Introduction: Despite careful control of environmental factors, there is a wide variety of interpersonal muscle-induced muscle damage. Genetic changes can also be important in determining susceptibility to injury. Weightlifting is one of the most successful Iranian sports teams in Olympics; so, the aim of this study was the investigation of the relationship between ACE I/D gene polymorphism rs4646994 and incidence and severity of sport injuries in Iranian male elite weightlifters. Materials and Methods: The study was cross-sectional research that conducted in 2022. The statistical population consisted of male elite weightlifters from Isfahan consisting of 31 athletes who all were investigated as sample. A questionnaire was used to collect the injuries data. Saliva samples were taken from all subjects and DNA was extracted and genotyped by using real-time polymerase chain reaction (real time-PCR). Chi-square and regression tests were used to analyze the data. Significance level was also considered at P <0.05. Results: There was a significant difference in the incidence of injuries in different genotypes. ID genotype (59.4%) had the highest incidence of injury in compare with DD (23.2%) and II (17.4%) genotypes (χ2 = 21.47, P = 0.001). Also, there was a significant difference in the severity of injuries in different genotypes (χ2 =15.59, P = 0.049); but there was no relationship between genotype and incidence and the severity of sports injuries in selected Iranian elite male weightlifters (P= 0.715). Conclusion: The results showed differences in the incidence and severity of sport injuries in different genotypes, hence it would be important in the prediction injury risk in weightlifters. [ABSTRACT FROM AUTHOR]

Prefractionation of complex mixtures of proteins derived from biological samples is indispensable for proteome analysis via top-down mass spectrometry (MS). Polyacrylamide gel electrophoresis (PAGE), which enables high-resolution protein separation based on molecular size, is a widely used technique in biochemical experiments and has the potential to be useful in sample fractionation for top-down MS analysis. However, the lack of a means to efficiently recover the separated proteins in-gel has always been a barrier to its use in sample prefractionation. In this study, we present a novel experimental workflow, called Passively Eluting Proteins from Polyacrylamide gels as Intact species for MS ("PEPPI-MS"), which allows top-down MS of PAGE-separated proteins. The optimization of Coomassie brilliant blue staining followed by the passive extraction step in the PEPPI-MS workflow enabled the efficient recovery of proteins, separated on commercial precast gels, from a wide range of molecular weight regions in under 10 min. Two-dimensional separation combining offline PEPPI-MS with online reversed-phase liquid chromatographic separation resulted in identification of over 1000 proteoforms recovered from the target region of the gel (≤50 kDa). Given the widespread availability and relatively low cost of traditional sodium dodecyl sulfate (SDS)-PAGE equipment, the PEPPI-MS workflow will be a powerful prefractionation strategy for top-down proteomics.

Colonies of the red harvester ant, Pogonomyrmex barbatus, regulate foraging activity based on food availability and local conditions. Colony variation in foraging behavior is thought to be linked to biogenic amine signaling and metabolism. Measurements of differences in neurotransmitters have not been made among ant colonies in a natural environment. Here, for the first time, we quantified tissue content of 4 biogenic amines (dopamine, serotonin, octopamine, and tyramine) in single forager brains from 9 red harvester ant colonies collected in the field. Capillary electrophoresis coupled with fast-scan cyclic voltammetry (CE-FSCV) was used to separate and detect the amines in individual ant brains. Low levels of biogenic amines were detected using field-amplified sample stacking by preparing a single brain tissue sample in acetonitrile and perchloric acid. The method provides low detection limits: 1 nM for dopamine, 2 nM for serotonin, 5 nM for octopamine, and 4 nM for tyramine. Overall, the content of dopamine (47 ± 9 pg/brain) was highest, followed by octopamine (36 ± 10 pg/brain), serotonin (20 ± 4 pg/brain), and tyramine (14 ± 3 pg/brain). Relative standard deviations were high, but there was less variation within a colony than among colonies, so the neurotransmitter content of each colony might change with environmental conditions. This study demonstrates that CE-FSCV is a useful method for investigating natural variation in neurotransmitter content in single ant brains and could be useful for future studies correlating tissue content with colony behavior such as foraging. Graphical abstract.

In an open microfluidic device, we investigate protein polyacrylamide gel electrophoresis (PAGE) separation performance on single-cell lysate. Single-cell protein electrophoresis is performed in a thin layer of polyacrylamide (PA) gel into which microwells are molded. Individual cells are isolated in these open microwells, then lysed on-chip with a dual lysis and electrophoresis sodium dodecyl sulfate (SDS) buffer. We scrutinize the effect of sieving gel composition on electromigration of protein targets, using a wide range of cellular protein standards (36 kDa to 289 kDa). We find that as PA concentration increases, protein electromigration deviates from the empirical log-linear relationship predicted between migration distance and molecular mass. We perform Ferguson analysis to calculate retardation coefficients and free solution mobilities of nine cellular protein standards and observe that the largest-molecular-mass protein, mTOR (289 kDa), does not behave as predicted by established linear-fit models for SDS-denatured proteins, indicating that mTOR is beyond the linear range of this assay. Lastly, we performed in-gel immunoprobing on the single-cell electrophoretic separations and observed that smaller pore-size gels (higher gel concentration) reduce protein diffusion out of the gel, which does not notably impact the measured immunoprobed protein expression. Compared to larger pore-size gels, the smaller pore-size gels lead to higher local concentrations of the target protein in each protein band, resulting in an increase in the signal-to-noise ratio (SNR) for each protein. Understanding the separation and immunoprobing performance at different gel concentrations improves assay design and optimization for target proteins.

For decades, there has been immense progress in miniaturizing analytical methods based on electrophoresis to improve sensitivity and to reduce sample volumes, separation times, and/or equipment cost and space requirements, in applications ranging from analysis of biological samples to environmental analysis to forensics. In the field of radiochemistry, where radiation-shielded laboratory space is limited, there has been great interest in harnessing the compactness, high efficiency, and speed of microfluidics to synthesize short-lived radiolabeled compounds. We recently proposed that analysis of these compounds could also benefit from miniaturization and have been investigating capillary electrophoresis (CE) and hybrid microchip electrophoresis (hybrid-MCE) as alternatives to the typically used high-performance liquid chromatography (HPLC). We previously showed separation of the positron-emission tomography (PET) imaging tracer 3'-deoxy-3'-fluorothymidine (FLT) from its impurities in a hybrid-MCE device with UV detection, with similar separation performance to HPLC, but with improved speed and lower sample volumes. In this paper, we have developed an integrated radiation detector to enable measurement of the emitted radiation from radiolabeled compounds. Though conventional radiation detectors have been incorporated into CE systems in the past, these approaches cannot be readily integrated into a compact hybrid-MCE device. We instead employed a solid-state avalanche photodiode (APD)-based detector for real-time, high-sensitivity β particle detection. The integrated system can reliably separate [18F]FLT from its impurities and perform chemical identification via coinjection with nonradioactive reference standard. This system can quantitate samples with radioactivity concentrations as low as 114 MBq/mL (3.1 mCi/mL), which is sufficient for analysis of radiochemical purity of radiopharmaceuticals.

Immunoassays and mass spectrometry are powerful single-cell protein analysis tools; however, interfacing and throughput bottlenecks remain. Here, we introduce three-dimensional single-cell immunoblots to detect both cytosolic and nuclear proteins. The 3D microfluidic device is a photoactive polyacrylamide gel with a microwell array-patterned face (xy) for cell isolation and lysis. Single-cell lysate in each microwell is "electrophoretically projected" into the 3rd dimension (z-axis), separated by size, and photo-captured in the gel for immunoprobing and confocal/light-sheet imaging. Design and analysis are informed by the physics of 3D diffusion. Electrophoresis throughput is > 2.5 cells/s (70× faster than published serial sampling), with 25 immunoblots/mm2 device area (>10× increase over previous immunoblots). The 3D microdevice design synchronizes analyses of hundreds of cells, compared to status quo serial analyses that impart hours-long delay between the first and last cells. Here, we introduce projection electrophoresis to augment the heavily genomic and transcriptomic single-cell atlases with protein-level profiling.

Doxorubicin (DOX) is one of the most widely used anticancer agents. DOX is known for inducing cardiotoxicity, resulting in the long-term development of heart failure. Intravascular delivery of DOX may benefit from the carriage by red blood cells (RBCs), as they can limit the systemic toxicity while delivering the DOX to the tumor. This study proposes a methodology for the synthesis of electrophoretically DOX-loaded red blood cells (RBC-DOX), as well as the assessment of its antitumorigenic effects in human colon cancer cells (HT-29), and in colon cancer xenograft models. In addition, healthy mice without tumors were dosed with RBC-DOX to assess cardiotoxicity via assessment of indexes of cardiac function after multiple doses of RBC-DOX. The HT-29 IC50 was found to be lower for RBC-DOX compared to free DOX. Tumor volume for the RBC-DOX group was smaller than the free DOX groups in HT-29 xenografts models. Statistically higher concentrations of DOX were found in the liver, spleen, and lungs for the RBC-DOX group compared to the free DOX group. However, the heart and the skin had statistically lower DOX concentrations for the RBC-DOX group compared to the free DOX group, with no significant differences in tumor biodistribution. All hemodynamic and cardiac function parameters were closer to control parameters for the RBC-DOX treated compared to for the free DOX-treated mice. These results suggest that RBC-DOX can be an alternative to prolong treatments with DOX, with superior antitumorigenic effects, decreased myelosuppression, and limited cardiac toxicity compared to equivalent doses of free DOX.

Introduction: Triticale (X. Triticosecale Wittmack) was created by doubling the number of F1 hybrid chromosomes between wheat (Triticum spp.) as the female parent and rye (Secale spp.) as the male parent. Investigation of the plants reaction to water shortage is systematically considered by identifying features that are related to drought tolerance subjected to the physiological, cellular, biochemical, and molecular bases. One of the main subjects regarding plants damage under drought stress is the accumulation of reactive oxygen species. Plants possess enzymatic and non-enzymatic systems to prevent oxygen free radical damage which are using them in an adaptive way under drought stress condition. Changes in the expressional level of antioxidant enzymes and the concentration of these enzymes, as well as increasing their activity, are among the very important changes occurring in the cellular level as the result of facing drought stress. Considering the role of the superoxide dismutase enzyme in regulating the amount of oxygen free radicals in plant cells, the aim of the present study was to investigate the gene expression of the superoxide dismutase enzyme with copper/zinc coenzyme under drought stress conditions.Materials and methods: Two genotypes of triticale (sensitive and tolerant) were subjected to drought stress conditions after establishment, and their aerial parts and roots were sampled at 0, 12, 36, and 72 hours after applying drought stress. Drought treatment was applied by weight based on the soil capacity of the pots and by daily weighing of the pots. Each time point repeated three times where the plant shoot was samples prior to plant root. This experiment was carried out in a factorial manner and with a completely randomized design in the greenhouse. The expression level of the superoxide dismutase gene with copper/zinc coenzyme was measured in the samples using the quantitative PCR method.Results: The results of the present study showed that the expression of the Cu/Zn-SOD gene is affected and changed under stress conditions. In general,, with the increase in the amount of stress severity and time of stress, the amount of immediate gene expression increased. The changes in the expression of this gene were greater in the shoots than in the roots, and the shoots responded faster to drought stress. Also, between the two sensitive and tolerant genotypes used in this experiment, the level of Cu/Zn-SOD gene expression in both roots and shoots was higher in the tolerant genotype than in the sensitive triticale genotype.Conclusion: According to the changes in expression and amount in two sensitive and tolerant triticale genotypes, it was generally determined that the superoxide dismutase enzyme is an essential enzyme in the plant to respond to drought stress and also to create tolerance in the triticale plant. Also, according to the changes in gene expression related to the Cu/Zn-SOD isozyme in triticale, it was determined that this isozyme is probably one of the active isozymes of the superoxide dismutase enzyme, whose expression is increased under drought stress conditions.

We attempt to study the evolution of the sickle cell anaemia in the region of Kinshasa by proposing three models based on Markov processes. The first two models namely, the idealistic and the quasi- idealistic models fail to completely describe the data sampled. The third model, the realistic one, leads to results that match the data collected and meets the predictions of serious institutions such as the World Health Organization (about 36 percent of the population belongs to the electrophoresis categories AS or SS).

Herbert Gritsch,1 Gerald Schrenk,1 Nina Weinhappl,1 Björn Mellgård,2 Bruce Ewenstein,2 Peter L Turecek3,4 1Pharmaceutical Sciences, Baxalta Innovations GmbH, a Takeda Company, Vienna, Austria; 2Rare Genetics and Hematology, Research & Development, Takeda Development Center Americas, Inc, Cambridge, MA, USA; 3Department of Pharmaceutical Sciences, Division of Pharmacology, University of Vienna, Vienna, Austria; 4Plasma Derived Therapies, Research & Development, Baxalta Innovations GmbH, a Takeda Company, Vienna, AustriaCorrespondence: Peter L Turecek, Baxalta Innovations GmbH, a Takeda company, DC-Tower, Donau-City-Straße 7, Vienna, A-1220, Austria, Tel +43 1 20 100 247 2520, Fax +43 1 20 100 247 5733, Email peter.turecek@takeda.comAbstract: Background: Recombinant von Willebrand factor (rVWF, vonicog alfa) is a purified VWF concentrate produced from Chinese hamster ovary cells. rVWF is not exposed to the VWF-cleaving protease ADAMTS13 and so is not subject to proteolytic degradation of large (L) and ultra-large (UL) VWF multimers by that enzyme.Purpose: To compare the structure and function of rVWF with the human plasma-derived VWF [pdVWF] concentrates Haemate P®/Humate-P®, Voncento®, Wilate®/Eqwilate®, and Wilfactin®/Willfact®; to investigate the relationship between VWF multimeric pattern and VWF:ristocetin cofactor (VWF:RCo) activity through population pharmacokinetic (PK) modeling in patients with severe von Willebrand disease (VWD) treated with rVWF.Methods: Analyses included VWF:RCo activity, VWF:collagen-binding activity, VWF:platelet glycoprotein Ib receptor binding, factor VIII (FVIII) binding capacity, and VWF-mediated platelet adhesion under flow conditions. VWF multimeric structure was determined by agarose gel electrophoresis. Population PK models describing the activity-time profile of small, medium, and L/UL multimers following intravenous administration of rVWF in patients with severe VWD were developed.Results: Findings demonstrate that rVWF contains a non-degraded VWF multimer pattern including the UL multimers not present in pdVWF concentrates. rVWF displayed higher specific platelet-binding activity, and faster mediation of platelet adhesion to collagen under shear stress versus pdVWF concentrates. rVWF also demonstrated higher FVIII binding capacity than Haemate P®, Voncento® and Wilate®. Modeling provided evidence that VWF:RCo activity in patients with severe VWD treated with rVWF is associated with L/UL VWF multimers in the circulation.Conclusions: Findings suggest that the L and UL multimers preserved in rVWF contribute to high biological activity and might be important for providing hemostatic efficacy.Keywords: agar, electrophoresis, factor VIII, plasma, von Willebrand diseases, von Willebrand factor

While conducting recombinant DNA technology procedures, such as DNA purification, agarose gel electrophoresis is often used for identification, characterization and quantification of DNA. The collection of data for experiments involving such techniques frequently involves capturing images using systems that are expensive and/or proprietary, such that they are not user-serviceable when they malfunction or become antiquated. In response to these limitations, work was done to replace the authors' existing aging Mac OS-based modular system with open-source software and generic hardware. Several versions of a modular imaging system that can be adjusted to fit nearly all use cases are described. The systems developed can accommodate diverse uses from research laboratories to educational environments where commercial systems could be unaffordable.

Background. In order to fully reveal the advantages of a cultivar and use it effectively in production, it is necessary to ensure high sowing and varietal qualities of seeds, which requires improvement in breeding and seed production. Biotechnological methods are increasingly used in initial seed production. High efficiency was shown by the method of electrophoresis of seed storage proteins – prolamins. The aim of this study was to assess the biotype composition and allelic state of the prolamin-coding loci in the new spring oat cultivar ‘Tobolyak’ for its further use in initial seed production.Materials and methods. In 2021, 1519 families of cv. ‘Tobolyak’ were studied using the method of native electrophoresis. Three grains from each family were selected for analyses by random sampling. Electrophoresis was carried out in vertical plates of 13.2% polyacrylamide gel at a constant voltage of 500 V for 4.5–5.0 hours.Results and conclusion. The study of cv. ‘Tobolyak’ families resulted in identifying 13 types of avenin patterns with a frequency of occurrence from 96.36 to 0.07%. A new block of avenin components controlled by the allele of the Avn B locus was described. Number 8 was assigned to this block. It was established that cv. ‘Tobolyak’ had been developed by crossing a minor biotype of cv. ‘Taezhnik’ (Avn 4.4.2) with the first biotype of cv. ‘Orion’ (Avn 2.8.2). The basis of cv. ‘Tobolyak’ was one genotype with the Avn 4.8.2 avenin formula. The remaining biotypes arose as a result of crossing the biotypes of cvs. ‘Taezhnik’ and ‘Orion’ as well as through mechanical contamination. To maintain the genetic stability of cv. ‘Tobolyak’, it is recommended to use only families of the first biotype in the process of original seed production.

LAMP assay is widely used for detecting pathogens. We observed that the conventional and gradient polymerase chain reaction (PCR) could not detect the extracted Escherichia coli DNA; real-time PCR was able to detect up to a certain limit (10−8 bacterial dilution). At the same time, the LAMP assay could detect the bacteria at a much lower concentration (10−14 dilution). The results of the LAMP assay were evaluated using agarose gel electrophoresis and DNA binding dye (PicoGreen), but only gel electrophoresis gave reliable results. Therefore, we propose using electrophoresis-based amplicon detection to overcome the limitations of dye-based detection. We believe that this amplicon detection will go a long way in the screening of potable drinking water.

Over the years, extrusion has been a multi-step thermal technique that has proven to be the most effective process to stabilize rice bran (RB). This study aimed to investigate the effects of extrusion treatment and temperature (15, 25, and 40 °C) on the storage stability, lipid oxidation, peroxidase, and peroxide values, free fatty acids, fatty acid composition, and protein variations of RB over 60 days. The study offers novel insights into the changes in RB's protein and amino acid compositions during extrusion and storage, which has not been extensively explored in prior research. After extrusion processing, peroxidase activity (POD) and lipase activity (LPS) were significantly reduced. However, peroxide value (PV), free fatty acids (FFA), and malondialdehyde content (MDA) observed a significantly increased by 0.64 mEqO2/kg, 8.3 mg/100 g, and 0.0005 μmol/L respectively. The storage stability of RB after extrusion shows that the POD, LPS, FFA, PV, and MDA were positively correlated with storage duration and temperature. The oleic acid/linoleic acid ratio in processed RB by extrusion had no significant changes during storage. The total and essential/non-essential amino acid ratios showed a downward trend of 5.26% and 8.76%, respectively. The first-order kinetics was the best-fitting model to describe the enzymatic inactivation and degradation of extruded RB during storage. The extrusion treatment did not affect the crude protein and the essential subunits of protein. Overall, the optimized extrusion procedure exhibited promising results in stabilizing the RB. [ABSTRACT FROM AUTHOR]

Background: Aging is associated with impaired renal function and structural alterations. Oxidative stress plays a vital role in renal senescence and damage. Sirtuin 1 (SIRT1) is thought to protect cells from oxidative stress through nuclear factor erythroid 2-related factor 2 (NRF2). Ellagic acid (EA), a natural antioxidant, has been demonstrated to have renoprotective roles in vitro and in vivo. This study investigated if SIRT1 and NRF2 mediate the protective effects of EA in aged kidneys. Methods: Male Wistar rats were divided into three groups including young (4 months), old, and old + EA (25 months). Young and old groups received EA solvent, while the old + EA group was treated with EA (30 mg/kg) by gavage for 30 days. Then, the level of renal oxidative stress, SIRT1 and NRF2 expression, kidney function parameters, and histopathological indices were measured. Results: Treatment with EA significantly increased the level of antioxidant enzymes and reduced malondialdehyde concentration (P < 0.01). Moreover, EA administration remarkably upregulated mRNA and protein levels of SIRT1 and NRF2 as well as deacetylated NRF2 protein (P < 0.05). Additionally, EA treated rats improved kidney function and histopathological scores (P < 0.05). Conclusions: These findings suggest that ellagic acid exerts protective effects on aged kidneys by activating SIRT1 and NRF2 signaling. [ABSTRACT FROM AUTHOR]

Zinc nanoparticles have a positive effect in enhancing growth and improving immunity, so this work aimed to investigate the role of Nano-zinc oxide as an immune stimulant and treatment of zinc deficient lambs in comparison with zinc in ordinary size. Thirty lambs were used and divided into three groups, the 1st group (N=10) control healthy group, the 2nd group (N=10) lambs suffered from zinc deficiency and treated with Nano-zinc oxide (nZnO), the 3rd group (N=10) lambs suffered from zinc deficiency treated with zinc oxide (ZnO) ordinary size. Serum samples were collected at zero-day (0-day), 7th, and 15th days of experiment. The serum analysis results of the zinc-deficient groups at 0-days revealed hypozincaemia, hypovitaminosis A and E, hypoproteinemia, hypoalbuminemia, hypoglobulinemia, decreased antioxidant (SOD, TAC, CAT, GSH) and immunoglobulin (γ2, γ1, β2, β1), while increased MDA, α2, and α1 compared to control healthy group. Most of parameters revealed rapid recovery from the 7th day of experiment in the nZnO group rather than the ZnO group. Some parameters of SOD, total protein, globulin, γ2, γ1, α2, and α1 appeared on the 7th day of experiment of the nZnO group significantly increased compared to the control healthy group, which revealed an improvement in the immune response. In conclusion; nZnO induced rapid recovery, and improved both immunity and antioxidants than zinc oxide ordinary sized. [ABSTRACT FROM AUTHOR]

Although breast cancer screening assays exist, many are inaccessible and have high turnaround times, leaving a significant need for better alternatives. Hypermethylation of tumor suppressor genes is a common epigenetic marker of breast cancer. Methylation tends to occur most frequently in the promoter and first exon regions of genes. Preliminary screening tests are crucial for informing patients whether they should pursue more involved testing. We selected RASSF1, previously demonstrated to be aberrantly methylated in liquid biopsies from breast cancer patients, as our gene of interest. Using CoBRA as our method for methylation quantification, we designed unique primer sets that amplify a portion of the CpG island spanning the 5′ end of the RASSF1 first exon. We integrated the CoBRA approach with a microfluidics-based electrophoresis quantification system (LabChip) and optimized the assay such that insightful results could be obtained without post-PCR purification or concentration, two steps traditionally included in CoBRA assays. Circumventing these steps resulted in a decreased turnaround time and mitigated the laboratory machinery and reagent requirements. Our streamlined technique has an estimated limit of detection of 9.1 ng/μL of input DNA and was able to quantify methylation with an average error of 4.3%. [ABSTRACT FROM AUTHOR]

Simple Summary: Natural products derived from plants have numerous clinical applications, including anti-cancer activity. In the present study, we identified three different plant extracts as strong inducers of cell death that were not reported previously. We focused on the most potent of these plants, Vernonanthura nudiflora (Vern). We demonstrated that the plant extracts obtained by treatment with a water and ethanol mixture killed tumor cells via multiple routes. These include impairing cell energy and metabolism, generating reactive oxygen species, increasing intracellular Ca2+, and inducing mitochondria-mediated apoptosis. We connected these activities to increased levels of the mitochondrial gatekeeper protein, VDAC1, which is associated with metabolism and apoptosis regulation. In a glioblastoma mouse model, Vern extract strongly inhibited tumor growth and induced massive tumor cell death, including cancer stem cells, by inhibiting blood supply and modulating the tumor microenvironment. The multipronged effects of hydroethanolic Vern extract make it a promising candidate for treating cancer. The mitochondrial voltage-dependent anion channel 1 (VDAC1) protein is involved in several essential cancer hallmarks, including energy and metabolism reprogramming and apoptotic cell death evasion. In this study, we demonstrated the ability of hydroethanolic extracts from three different plants, Vernonanthura nudiflora (Vern), Baccharis trimera (Bac), and Plantago major (Pla), to induce cell death. We focused on the most active Vern extract. We demonstrated that it activates multiple pathways that lead to impaired cell energy and metabolism homeostasis, elevated ROS production, increased intracellular Ca2+, and mitochondria-mediated apoptosis. The massive cell death generated by this plant extract's active compounds involves the induction of VDAC1 overexpression and oligomerization and, thereby, apoptosis. Gas chromatography of the hydroethanolic plant extract identified dozens of compounds, including phytol and ethyl linoleate, with the former producing similar effects as the Vern hydroethanolic extract but at 10-fold higher concentrations than those found in the extract. In a xenograft glioblastoma mouse model, both the Vern extract and phytol strongly inhibited tumor growth and cell proliferation and induced massive tumor cell death, including of cancer stem cells, inhibiting angiogenesis and modulating the tumor microenvironment. Taken together, the multiple effects of Vern extract make it a promising potential cancer therapeutic. [ABSTRACT FROM AUTHOR]

Objectives: In this study, we aimed to investigate the therapeutic effect of anti-receptor activator of nuclear factor kappa-kB ligand (RANKL) monoclonal antibodies R748-1-1-1, R748-1-1-2 and R748-1-1-3 on rheumatoid arthritis (RA) in a rat model. Materials and methods: Gene cloning, hybridoma technology, affinity purification, enzyme-linked immunosorbent assay, general observation, hematoxylin-eosin staining, X-ray, and many other experimental techniques were used in this study. Results: Improved collagen-induced arthritis (CIA) modeling was successfully constructed. The RANKL gene was cloned and the anti-RANKL monoclonal antibody was prepared. Following treatment with the anti-RANKL monoclonal antibody, the soft tissue swelling of the hind paws, the joint thickening, the narrowed joint gap, and the blurred edge of the bone joint were improved. The pathological changes such as synovial hyperplasia of fibrous tissue, cartilage and bone destruction were significantly decreased in the anti-RANKL monoclonal antibody-treated CIA group. Compared to the normal control group and phosphate buffer saline (PBS)-treated CIA group, the expression of tumor necrosis factor-alpha (TNF-a) and interleukin-1 (IL-1) in antibody-treated CIA group, positive drug-treated CIA group, and IgG-treated CIA group were decreased (p<0.05). Conclusion: The anti-RANKL monoclonal antibody can promote the therapeutic effect of RA rats, indicating that the anti-RANKL monoclonal antibody has a certain potential value and may be beneficial to the further study of the mechanism of RA treatment. [ABSTRACT FROM AUTHOR]

Protein analysis is an important approach for disease diagnosis, in which sample pretreatment is an essential step since protein samples are often complex and many protein biomarkers are of low abundance. Here, given the good openness and light transmission of liquid plasticine (LP), which is a liquid entity formed by SiO2 nanoparticles and encapsulated aqueous solution, we developed a LP‐based field‐amplified sample stacking (FASS) system for protein enrichment. The system was composed of a LP container, a sample solution and a Tris‐HCl solution containing hydroxyethyl cellulose (HEC). The system design, mechanism investigation, optimization of experimental parameters and characterization of LP‐FASS performance for protein enrichment were well studied. Under the optimized experimental conditions of 1 % HEC, 100 mm Tris‐HCl and 100 V in the LP‐FASS system, a 40–80 times enrichment of proteins was obtained in 40 min using bovine hemoglobin (BHb) as the model protein using the constructed LP‐FASS system. The simultaneous enrichment of multiple proteins (phycocyanin, BHb and cytochrome C) was also realized using the system. The LP‐FASS system can serve as a new platform for protein enrichment which is easy to be combined with online and offline detections. [ABSTRACT FROM AUTHOR]