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Aggregation of protein-based therapeutics can occur during development, production, or storage and can lead to loss of efficacy and potential toxicity. Native mass spectrometry of a covalently linked pentameric monoclonal antibody complex with a mass of ∼800 kDa reveals several distinct conformations, smaller complexes, and abundant higher-order aggregates of the pentameric species. Charge detection mass spectrometry (CDMS) reveals individual oligomers up to the pentamer mAb trimer (15 individual mAb molecules; ∼2.4 MDa) whereas intermediate aggregates composed of 6-9 mAb molecules and aggregates larger than the pentameric dimer (1.6 MDa) were not detected/resolved by standard mass spectrometry, size exclusion chromatography (SEC), capillary electrophoresis (CE-SDS), or by mass photometry. Conventional quadrupole time-of-flight mass spectrometry (QTOF MS), mass photometry, SEC, and CE-SDS did not resolve partially or more fully unfolded conformations of each oligomer that were readily identified using CDMS by their significantly higher extents of charging. Trends in the charge-state distributions of individual oligomers provides detailed insight into how the structures of compact and elongated mAb aggregates change as a function of aggregate size. These results demonstrate the advantages of CDMS for obtaining accurate masses and information about the conformations of large antibody aggregates despite extensive overlapping m/z values. These results open up the ability to investigate structural changes that occur in small, soluble oligomers during the earliest stages of aggregation for antibodies or other proteins.

Background: Plants that contain brewing microorganisms are used in traditional fermentation starters, which are an essential part of local diet, nutrition, life, and health. Regionally, the plant species used and the microorganisms included in traditional fermentation starters are diverse, endowing local fermented drinks with different flavors and health benefits. However, related traditional knowledge has been scarcely documented or revealed. Methods: An ethnobotanical survey was conducted in five towns of Nayong County in northwestern Guizhou, China. Snowball sampling, semi-structured interviews, free lists, and participatory observation were used to collect information on Jiuqu Plants (JPs) and jiuqu-making techniques. The PacBio platform was used to study the microbial community structure and diversity in the Chuanqing people's jiuqu. Results: In total, 225 informants were interviewed, including 116 who provided plants and technological processes for making Chinese baijiu jiuqu (CBJ) and 139 who provided information about making fermented glutinous rice jiuqu (FGRJ). This study found that older people have more abundant knowledge about CBJ plants. Poaceae was found to be the dominant family used in making CBJ and FGRJ (7 species each). Compared to individual plant parts, the whole plant is most commonly used in two kinds of jiuqu (19.5% in CBJ and 22.6% in FGRJ). The Chuanqing people's jiuqu is used to treat dietary stagnation and indigestion. The highest relative frequency of citation of the CBJ plant was Ficus tikoua Bureau, and the counterpart of the FGRJ plant was Buddleja macrostachya Benth. The dominant bacterial species in jiuqu were Gluconobacter japonicus (YQ1, YQ4) and Pediococcus pentosaceus (YQ2, YQ3), and the dominant fungal species was Rhizopus oryzae. Conclusion: For the first time, this study documents the unique traditional jiuqu knowledge and reveals the microbial mystery behind the FGRJ of the Chuanqing people. Therefore, this study encourages the use of online social media platforms in order to spread Jiuqu culture, the use of the new media wave in order to create multimedia databases, and also suggests that local communities should develop preservation intervention programs, in addition to nurturing the inheritors in order to prevent the disappearance of traditional Jiuqu knowledge. This research contributes to the conservation and demystification of the traditional jiuqu knowledge of the Chuanqing people and lays the foundation for further research on its microbiology, nutrition, and metabolomics. [ABSTRACT FROM AUTHOR]

This work was devoted to a study of the composition of protein concentrates from amaranth grain (Amaranthus hypochondriacus L.) of variety Voronezh. Amaranth protein concentrates were obtained by alkaline extraction of proteins and neutralization of the solution followed by ultrafiltration, by separating the starch fraction with amylolytic enzymes, and by alkaline extraction of proteins and their precipitation at pH 4.5. Conditions for the extraction of proteins followed by chromatography–mass spectrometric analysis and identification were selected. It was found that proteins from amaranth grain were more effectively extracted with a buffer with urea at protein concentrations of 1.7, 1.9, and 2.9 mg/cm3 in solution, respectively, while a buffer with detergents was more effective for the extraction of low-molecular-weight proteins at protein concentrations of 4.9, 2.9, and 9.0 mg/cm3 in solution, respectively. As a result of HPLC–MS/MS analysis followed by identification and search in the UNIPROT database, it was established that the main protein of amaranth grain is 11S-globulin, which acts as a reserve protein of amaranth seeds. In amaranth concentrates, 14 unique proteins belonging only to A. hipochondriacus L. were identified, and also proteins that did not belong to this species were reliably identified. Based on the results of semiquantitative analysis of the peptide profile of amaranth grain protein concentrates, a high frequency of occurrence of the main 11S-globulin proteins was established in all samples. The frequency of occurrence of other proteins in samples obtained by different methods differed significantly due to the peculiarities of protein isolation from amaranth grain. The results obtained can be used to prepare plant protein concentrates with a given protein composition. [ABSTRACT FROM AUTHOR]

Peripheral neuropathy is a prevalent complication in plasma cell disorders, posing significant diagnostic and therapeutic challenges. This study presents three cases initially diagnosed with chronic inflammatory demyelinating polyneuropathy (CIDP). Despite initial symptom regression post-immunomodulatory treatment, the patients exhibited progressive neurological deficits. Advanced laboratory evaluation confirmed monoclonal protein presence, yet traditional diagnostic methods, including bone marrow biopsy and flow cytometry, yielded normal results. Utilizing 18F-FDG PET/CT, we identified multiple hypermetabolic vertebral lesions, which upon biopsy, confirmed the diagnosis of plasmacytoma. Our findings underscore the utility of PET/CT as a reliable diagnostic tool for monoclonal gammopathy associated neuropathy, advocating for its consideration in cases with equivocal diagnosis. When the diagnosis is in doubt, biopsy of a lesion may facilitate early and accurate diagnosis, potentially influencing treatment strategies and patient outcomes. [ABSTRACT FROM AUTHOR]

The red V2O5 sol was prepared by facile alternating stirring and ultrasonic dispersion route. A V2O5 film fabricated on ITO-PET by electrophoresis method was used to demonstrate a flexible electrochromic device. The effects of applied voltage and electrophoresis time for V2O5films were investigated in detail. When the electrophoresis time was kept 30 s, the applied voltage was above 3 V to deposit uniform V2O5-PET film. The electrophoresis time could more significantly regulate the deposition of V2O5 on ITO-PET film. A flexible electrochromic device with "sandwich" structure was fabricated using V2O5 film, and its electrochromic performance of the device was evaluated. The flexible V2O5 electrochromic devices show the multicolor electrochromic performance of yellow, yellow-green, green and orange-red. The coloration efficiency values of V2O5 devices were 21.6 cm2C − 1 (from initial yellow to green) and 26.9 cm2C − 1 (from green to orange-red), respectively, and the highest transmittance modulation range was 51% at 741 nm. The work provided a facile and green method for fabrication of flexible V2O5 electrochromic device. [ABSTRACT FROM AUTHOR]

The ion steric repulsion and ion-solvent interactions, due to the finite size of ions, are analyzed for electrophoresis of a droplet in a non-dilute electrolyte in which the ionic volume fraction is O (0.01). In this study, the viscosity of the suspension medium is considered to be dependent on the local ionic volume fraction. The electrochemical potential of the finite-sized ions is modified to take into account the steric interactions. The impact of the Marangoni stress due to the non-uniform distribution of the non-ionized surfactant is taken into account. Governing equations are solved under a weak applied field consideration. At a lower range of surface charge density, the mobility is found to increase with the droplet viscosity, in which the Marangoni stress is found to enhance the mobility. The counterion saturation created by the ion steric interaction attenuates the screening of the surface charge and hence, enhances the mobility. However, the enhanced viscosity of the medium for the non-dilute electrolyte reduces the mobility and this reduction is higher in a monovalent electrolyte as compared to a multivalent electrolyte. The standard model shows a singularity in mobility when the surface potential is varied. However, the dependence of mobility on surface potential in the modified model is found to exhibit a smooth variation. • Modeling electrophoresis of a surfactant-laden highly charged viscous droplet. • Standard model is extended to include the finite ion size effects. • Ion steric interactions and dependence of viscosity on ionic concentration is included. • Marangoni stress due to surfactant concentration gradient is analyzed. • The limitation of the point-charge based standard model is highlighted. [ABSTRACT FROM AUTHOR]

1,2-propylene glycol and its 50-50 w/w mixture with water were used to prepare heat transfer fluids based on amino-functionalized silica. On top of pH-neutral dispersions (no reagents added except for the solvent and the particles), dispersions acidified with acetic acid and with HCl were used to enhance the positive electric charge of silica particles. The colloidal particles had a positive zeta potential >40 mV and showed apparent particle radii of 70 nm, and these properties remained unchanged on heating up to 80 °C for up to 28 days. [ABSTRACT FROM AUTHOR]

A general theory is developed for the time dependent transient electrophoretic mobility of spherical colloidal particles in a salt-free liquid medium containing only counterions when a step external electric field is suddenly applied to the colloidal suspension. It is found that as in the case of the steady electrophoretic mobility in a salt-free medium, there is a certain critical value of the particle surface charge separating two cases, that is, the low-surface-charge case and the high-surface-charge case. In the latter case the counterion condensation takes place near the particle surface. For the low-surface charge case, the transient electrophoretic mobility agrees with that of a sphere in an electrolyte solution in the limit of very low electrolyte concentrations. For the high-surface-charge case, however, the transient mobility becomes independent of the particle surface charge because of the counterion condensation effects. A simple expression is derived for the ratio of the transient electrophoretic mobility to the steady electrophoretic mobility, which is found to take the same form irrespective of the magnitude of the particle surface charge. Using this equation, it is now possible to predict how the system will approach its final steady state. [ABSTRACT FROM AUTHOR]

Electronic paper displays rely on electrokinetic effects in nonpolar solvents to drive the displacement of colloidal particles within a fluidic cell. While Electrophoresis (EP) is a well-established and frequently employed phenomenon, electro-osmosis (EO), which drives fluid flow along charged solid surfaces, has not been studied as extensively. We hypothesize that by exploiting the interplay between these effects, an enhanced particle transport can be achieved. In this study, we experimentally investigate the combined effects of EP and EO for colloidal particles in non-polar solvents, driven by an electric field. We use astigmatism micro-particle tracking velocimetry (A- μ PTV) to measure the motion of charged particles within model fluidic cells. Using a simple approach that relies on basic fluid flow properties we extract the contributions due to EP and EO, finding that EO contributes significantly to particle transport. The validity of our approach is confirmed by measurements on particles with different magnitudes of charge, and by comparison to numerical simulations. We find that EO flows can play a dominant role in the transport of particles in electrokinetic display devices. This can be exploited to speed up particle transport, potentially yielding displays with significantly faster switching times. Interplay of electrokinetic effects : Driven by the applied electric field, both electro-osmosis (EO, top left), which drives a flow along the surfaces of the fluidic cell, and electrophoresis (EP, top right), which drives a migration of particles relative to the background liquid, occur in our system. A typical example of the resultant particle velocity field (bottom) illustrates the important role of EO on particle transport in the fluidic cell. • Studied electrokinetics of particles in nonpolar solvent using astigmatism micro-PTV. • Both electrophoresis (EP) and electro-osmosis (EO) play important roles. • Combination of EO and EP could be exploited in faster switching e-paper displays. [ABSTRACT FROM AUTHOR]

Chirality is a fundamental and ubiquitous property of nature involved in multiple fields of science. In particular, the possible resolution of the enantiomeric forms of a molecule is crucial in the pharmaceutical, food, and agrochemical industries. The search for efficient, broad-spectrum, and yet simple methods for obtaining enantiomerically pure substances is a current challenge. Enantioselective resolution methods rely on an asymmetric environment that allows the two antipodes of a chiral molecule to be distinguished. In addition to enantiomeric separation techniques, such as chromatography and electrophoresis, new promising approaches involving out-of-the-scheme synergistic effects between chiral selectors (CS) and external stimuli are emerging. This Trends article discusses different enantioselective mechanisms triggered by unconventional physicochemical stimuli for the design of avant-garde approaches that could offer novel perspectives in the field of chiral resolution. [ABSTRACT FROM AUTHOR]

Introduction: Since its discovery in 1961, methicillin-resistant Staphylococcus aureus (MRSA) has been recognized as a significant healthcare-associated pathogen (HA-MRSA) and a notorious 'superbug'. Typing is crucial for surveillance, epidemiology analysis, infection control of MRSA and sequencing of the spa gene is one of the most common methods used for determining the origin of this bacterium in humans and animals. This research aimed to determine the antibiotic resistance and spa type of S. aureus strains collected from outpatients in two hospitals in the Wasit province of Iraq. Material & Methods: The study analyzed 200 outpatient MRSA isolates by collecting nasal and sputum samples from patients. Standard biochemical and molecular methods based on the nuc gene were used to identify S. aureus bacteria and amplify the mecA and spa genes. The Kirby-Bauer disc diffusion method was employed to determine the antibiotic sensitivity of the isolates using penicillin, cefoxitin, vancomycin, gentamicin, erythromycin, tetracycline, imipenem, clindamycin, chloramphenicol and rifampicin. Results: Methods. The prevalence of MRSA was more common in women than in men. Antibiogram results showed that most of the isolates were resistant to penicillin (94.2%) and sensitive to imipenem (100%), clindamycin (100%), and chloramphenicol (100%). Of these 35 isolates, 30 (87.5%) and 26 strains (74.3%) were positive for the mecA and spa genes. Typing based on spa gene sequencing revealed four different patterns: t386, t3579, U0002 and U0234. Conclusion: Variations in the spa gene among different S. aureus isolates may he of clinical importance when treating staphylococcal infections. In this study, spa typing revealed four different patterns in Iraq, representing diagnostic and therapeutic implications. [ABSTRACT FROM AUTHOR]

The objective of this research was to identify the morphology and molecular profile of crab caught from the coast of Tambala Village, Tombariri District, Minahasa Regency, North Sulawesi. Crabs were collected from the intertidal zone at night during the lowest tide. Crabs were morphological identified by description of shape, color and size of the carapace. Molecular identification was done through DNA barcoding including DNA extraction, amplification of the cytochrome c oxidase subunit I (COI) gene and electrophoresis. Morphological and genetic analysis identified the crab species as Metopograpsus oceanicus.

Background: Deoxyribonucleic acid (DNA) molecules can be effectively visualized when stained and observed under ultraviolet light during the electrophoresis process. The commonly used dye Ethidium bromide (EtBr) is considered hazardous due to its potential to cause mutations, cancer, and congenital disabilities. Various alternative dyes have been reported, one of which is hematoxylin. Hematoxylin compounds do not have mutagenic potential and are easier to apply than EtBr. However, there is no optimal variation in concentration and duration of staining for clear and effective visualization of DNA bands. Purpose: To find the concentration and staining time of harris hematoxylin staining for DNA bands from agarose gel electrophoresis. Method: An experimental method with a group comparison statistical design. The amplified DNA I6S rRNA gene from Escherichia coli (584 bp), which had undergone electrophoresis, was stained using harris hematoxylin dye at 0.01%; 0.02%; 0.03%; 0.04%; and 0.05% concentrations and immersion times soaking times of 10, 15, 20, and 30 minutes variations. The intensity of the DNA bands was analyzed using ImageJ. The staining power of the experimental groups was compared to the intensity of control dye and given a grading score of 1 - 4. The experiment was repeated twice, and the mean grading score was calculated. The highest mean value was considered the most optimal value. Result: A concentration of 0.02% showed relatively constant staining intensity for each soaking time. A mean value of 3.5 was obtained for a 0.01% concentration for 15 minutes. A 0.03% and 0.04% concentrations for 20 minutes. Conclusion: The highest mean value of 4 was obtained for Harris hematoxylin at 0.05% for 15 minutes.

The effect of four meat thawing techniques including chill temperature, room temperature, tap water and microwave on quality characteristics, lipid oxidation, proteins degradation, microbial spoilage and the microstructure of beef longissimus lumborum muscle were compared. A fresh portion of longissimus lumborum muscle was obtained from eight beef carcasses after 24 h postmortem. The findings indicated that all of the thawed samples, particularly the microwave beef samples, had a markedly worse quality than those of fresh meat. Among the thawing methods, changes in the beef quality of the chill temperature samples were significantly less. The increases in malondialdehyde value and decreases in free thiol content indicated that proteins and lipids in the beef samples subjected to room temperature, tap water, and microwave thawing methods were oxidized while thawed beef at a chill temperature did not significantly affect the lipid-protein oxidation compared to fresh beef. The greater myosin degradation was observed in room temperature, tap water and microwave thawing treatments, while actin degradation was not affected. Higher growth of total bacterial counts was presented by samples subjected to tap water and at room temperature thawing compared to microwave and chill temperature thawing. The sensory traits of the chiller thawing method were better than those of other methods. Images from scanning electron microscopy demonstrated that the myofibril configurations in the thawing samples were more loose than in the fresh beef samples, which had a compact and organized structure. This was demonstrated by the thawing samples' evidently higher myofibril gap value.

Abstract Introduction Patients with psoriatic arthritis have some lipid metabolism changes and higher risk of metabolic syndrome (MetS) and cardiovascular diseases, regardless of traditional risk factors, suggesting that chronic inflammation itself plays a central role concerning the atherosclerosis. However, there is a lack of information regarding atherogenic pattern and lipoprotein subfractions burden in these individuals. Aim To evaluate the HDL and LDL-cholesterol plasmatic levels and their subfractions after a nutritional intervention in patients with psoriatic arthritis (PsA). Methods This was a randomized, placebo-controlled clinical trial of a 12-week nutritional intervention. PsA patients were randomly assigned to 1-Placebo: 1 g of soybean oil daily, no dietetic intervention; 2-Diet + Supplementation: an individualized diet, supplemented with 604 mg of omega-3 fatty acids, three times a day; and 3-Diet + Placebo: individualized diet + 1 g of soybean oil. The LDL subfractions were classified as non-atherogenic (NAth), atherogenic (Ath) or highly atherogenic (HAth), whereas the HDL subfractions were classified as small, medium, or large particles, according to the current recommendation based on lipoproteins electrophoresis. Results A total of 91 patients were included in the study. About 62% of patients (n = 56) had an Ath or HAth profile and the main risk factors associated were male gender, longer skin disease duration and higher BMI. Thirty-two patients (35%) had a high-risk lipoprotein profile despite having LDL plasmatic levels below 100 mg/dL. The 12-week nutritional intervention did not alter the LDL subfractions. However, there were significant improvement of HDL subfractions. Conclusion Recognizing the pro-atherogenic subfractions LDL pattern could be a relevant strategy for identifying PsA patients with higher cardiovascular risk, regardless total LDL plasmatic levels and disease activity. In addition, a short-term nutritional intervention based on supervised and individualized diet added to omega-3 fatty acids changed positively the HDLLARGE subfractions, while LDLLARGE subfraction was improved in hypercholesterolemic individuals. ClinicalTrials.gov identifier: NCT03142503 ( http://www.clinicaltrials.gov/ ).

The study of genetic resources using prolamin polymorphism in wheat cultivars from countries with different climatic conditions makes it possible to identify and trace the preference for the selection of the alleles of gliadine-coding loci characteristic of specific conditions. The aim of the study was to determine the “gliadin profile” of the collection of common wheat (Triticum aestivum L.) from breeding centers in Russia and Kazakhstan by studying the genetic diversity of allelic variants of gliadin-coding loci. Intrapopulation (μ ± Sμ) and genetic (H) diversity, the proportion of rare alleles (h ± Sh), identity criterion (I) and genetic similarity (r) of common wheat from eight breeding centers in Russia and Kazakhstan have been calculated. It has been ascertained that the samples of common wheat bred in Kostanay region (Karabalyk Agricultural Experimental Station, Kazakhstan) and Chelyabinsk region (Chelyabinsk Research Institute of Agriculture, Russia) had the highest intrapopulation diversity of gliadin alleles. The proportion of rare alleles (h) at Gli-B1 and Gli-D1 loci was the highest in the wheat cultivars bred by the Federal Center of Agriculture Research of the South-East Region (Saratov region, Russia), which is explained by a high frequency of occurrence of Gli-B1e (86 %) and Gli-D1a (89.9 %) alleles. Based on identity criterion (I), the studied samples of common wheat from different regions of Kazakhstan and Russia have differences in gliadincoding loci. The highest value of I = 619.0 was found when comparing wheat samples originated from Kostanay and Saratov regions, and the lowest I = 114.4, for wheat cultivars from Tyumen and Chelyabinsk regions. Some region-specific gliadin alleles in wheat samples have been identified. A combination of Gli-A1f, Gli-B1e and Gli-Da alleles has been identified in the majority of wheat samples from Kazakhstan and Russia. Alleles (Gli-A1f, Gli-A1i, Gli-A1m, Gli-A1o, Gli-B1e, Gli-D1a, Gli-D1f, Gli-A2q, Gli-B2o, and Gli-D2a) turned out to be characteristic and were found with varying frequency in wheat cultivars in eight regions of Russia and Kazakhstan. The highest intravarietal polymorphism (51.1 %) was observed in wheat cultivars bred in Omsk region (Russia) and the lowest (16.6 %), in Pavlodar region (Kazakhstan). On the basis of the allele frequencies, a “gliadin profile” of wheat from various regions and breeding institutions of Russia and Kazakhstan was compiled, which can be used for the selection of parent pairs in the breeding process, the control of cultivars during reproduction, as well as for assessing varietal purity.

Abstract Background Electrotransfection is based on application of high-voltage pulses that transiently increase membrane permeability, which enables delivery of DNA and RNA in vitro and in vivo. Its advantage in applications such as gene therapy and vaccination is that it does not use viral vectors. Skeletal muscles are among the most commonly used target tissues. While siRNA delivery into undifferentiated myoblasts is very efficient, electrotransfection of siRNA into differentiated myotubes presents a challenge. Our aim was to develop efficient protocol for electroporation-based siRNA delivery in cultured primary human myotubes and to identify crucial mechanisms and parameters that would enable faster optimization of electrotransfection in various cell lines. Results We established optimal electroporation parameters for efficient siRNA delivery in cultured myotubes and achieved efficient knock-down of HIF-1α while preserving cells viability. The results show that electropermeabilization is a crucial step for siRNA electrotransfection in myotubes. Decrease in viability was observed for higher electric energy of the pulses, conversely lower pulse energy enabled higher electrotransfection silencing yield. Experimental data together with the theoretical analysis demonstrate that siRNA electrotransfer is a complex process where electropermeabilization, electrophoresis, siRNA translocation, and viability are all functions of pulsing parameters. However, despite this complexity, we demonstrated that pulse parameters for efficient delivery of small molecule such as PI, can be used as a starting point for optimization of electroporation parameters for siRNA delivery into cells in vitro if viability is preserved. Conclusions The optimized experimental protocol provides the basis for application of electrotransfer for silencing of various target genes in cultured human myotubes and more broadly for electrotransfection of various primary cell and cell lines. Together with the theoretical analysis our data offer new insights into mechanisms that underlie electroporation-based delivery of short RNA molecules, which can aid to faster optimisation of the pulse parameters in vitro and in vivo.

Abstract Copper-doped ZnO nanoparticles with the formula Zn1−x(Cu)O, where x = 0.0, 0.03, 0.05, and 0.07 were produced using the co-precipitation process. Physical, chemical, and structural properties were properly examined. Powdered X-ray diffraction (P-XRD) patterns revealed the formation of hexagonal wurtzite crystal structure in all samples, through atomic substitutional incorporation in the Cu-doped ZnO lattice. The presence of Cu ions and their dissolution in the host ZnO crystal structure was supported by FT-IR spectra. HR-TEM images were used to assess the average size, morphology, and shape regularity of the synthesized samples. The form and homogeneity of the ZnO changed when Cu ions were substituted, as evidenced by FE-SEM/EDX analysis. The presence of copper signals in the Cu-doped samples indicates that the doping was successful. The decrease in zeta potential with an increased copper doping percentage designates that the nanoparticles (NPs) are more stable, which could be attributed to an increase in the ionic strength of the aqueous solution. The synthesized NPs were evaluated for their substantial in vitro antioxidant properties. In addition, the antimicrobial efficacy of the materials was tested against pathogenic microorganisms. Regarding the anti-diabetic activity, the 7Cu ZnO sample showed the highest inhibitory effect on the α-amylase enzyme. No variations were observed in the activities of the acetylcholinesterase enzyme (AChE) and proteinase enzymes with ZnO and samples doped with different concentrations of Cu. Therefore, further studies are recommended to reveal the in-vitro anti-diabetic activity of the studied doped samples. Finally, molecular docking provided valuable insights into the potential binding interactions of Cu-doped ZnO with α-amylase, FabH of E. coli, and Penicillin-binding proteins of S. aureus. These outcomes suggest that the prepared materials may have an inhibitory effect on enzymes and hold promise in the battle against microbial infections and diabetes.

Dermatophytes are the keratinophilic fungi which infect humans and is the most recurring type of disease. The high level of transmissibility creates an epidemiological risk and emphasises the significance of these illnesses. However, a growing number of reports describing dermatophytes can cause deep infections in diabetic and immunocompromised patients, by invading deep layers like the dermis and hypodermis. Despite the prevalence and significance of dermatophytes in clinical mycology, it is not always possible to accurately diagnose this specific infection due to its overlapping structures among species of dermatophytes. Since it is difficult to identify species that exhibit weak characteristics in the morphological highlights, identification of the dermatophyte is often relied on its morphological analysis, which is a laborious process and demands skill. The massive shift in genetic variation, the source of infection, and epidemiological research can be discovered using molecular approaches. Therefore, the development of an accurate laboratory test for dermatophyte species identification is essential for the prevention and efficient management of dermatophytoses. One such methodology allows use of PCR technology which has many methods for molecular level characterization which is rapid, efficient, and capable of producing DNA polymorphisms specific to various dermatophyte species based on distinctive band patterns seen by agarose gel electrophoresis. The RAPD-PCR approach will be used in this study protocol to molecularly characterize the dermatophytes for precise speciation of the sample. In addition to improving knowledge of fungal biology and pathology with a focus on adaptive mechanisms to combat difficult conditions from host counteractions, there is a need to improve awareness of the importance of these diseases through accurate epidemiological data. The advantages of molecular approaches for characterizing objects over traditional methods are their sensitivity and specificity.

The analysis of dietary supplements is far less regulated than pharmaceuticals, leading to potential quality issues. Considering their positive effect, many athletes consume supplements containing l-histidine and β-alanine. A new microfluidic method for the determination of l-histidine and β-alanine in dietary supplement formulations has been developed. For the first time, capacitively coupled contactless conductivity detection was employed for the microchip electrophoresis of amino acids in real samples. A linear relationship between detector response and concentration was observed in the range of 10–100 µmol L–1 for l-histidine (R2 = 0.9968) and β-alanine (R2 = 0.9954), while achieved limits of detection (3 × S/N ratio) were 4.2 µmol L–1 and 5.2 µmol L–1, respectively. The accuracy of the method was confirmed using recovery experiments as well as CE-UV-VIS and HPLC-UV-VIS techniques. The developed method allows unambiguous identification of amino acids in native form without chemical derivatization and with the possibility of simultaneous analysis of amino acids with metal cations. [ABSTRACT FROM AUTHOR]

Background: Cervical cancer is one of the most common gynecological malignancies. Previous studies have shown that the ethanol extract of Sophora moorcroftiana seeds (EESMS) possesses an antiproliferative effect on several tumors in vitro. Therefore, in this study, we assessed the impact of EESMS on human cervical carcinoma (HeLa) cell proliferation. Methods: The proliferation and apoptotic effects of HeLa cells treated with EESMS were evaluated using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide assay, dual acridine orange/ethidium bromide double staining, flow cytometry, and western blotting. Single-cell level atomic force microscopy (AFM) was conducted to detect the mechanical properties of HeLa cells, and proteomics and bioinformatics methods were used to elucidate the molecular mechanisms of EESMS. Results: EESMS treatment inhibited HeLa cell proliferation by blocking the G0/G1 phase, increasing the expression of Caspase-3 and affecting its mechanical properties, and the EESMS indicated no significant inhibitory effect on mouse fibroblasts L929 cell line. In total, 218 differentially expressed proteins were identified using two-dimensional electrophoresis, and eight differentially expressed proteins were successfully identified using matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry. The differentially expressed proteins were involved in various cellular and biological processes. Conclusion: This study provides a perspective on how cells change through biomechanics and a further theoretical foundation for the future application of Sophora moorcroftiana as a novel low-toxicity chemotherapy medication for treating human cervical cancer. [ABSTRACT FROM AUTHOR]

Copyright of Anbar Journal of Agricultural Sciences is the property of Republic of Iraq Ministry of Higher Education & Scientific Research (MOHESR) 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.)

DNA knots are deleterious for living cells if not removed. Several theoretical and simulation approaches address the question of how topoisomerases select the intermolecular passages that preferentially lead to unknotting rather than to the knotting of randomly fluctuating DNA molecules, but the formation of knots in vivo remains poorly understood. DNA knots form in vivo in non-replicating and replicating molecules, and supercoiling as well as intertwining are thought to play a crucial role in both the formation and resolution of DNA knots by topoisomerase IV. To confirm this idea, we used two-dimensional agarose gel electrophoresis run with different concentrations of chloroquine to demonstrate that non-replicating pBR322 plasmids grown in a topoisomerase I-defective E. coli strain (RS2λ) were more negatively supercoiled than in a wild-type strain (W3110) and, concurrently, showed significantly fewer knots. In this way, using wild-type and E. coli mutant strains, we confirmed that one of the biological functions of DNA supercoiling is to reduce the formation of DNA knots. [ABSTRACT FROM AUTHOR]

Dental implants offer a reliable solution for replacing damaged tooth roots. This research investigates the comparative performance of acetone and methanol as suspension media in the fabrication of stainless steel type 316L-based dental implants using the Electrophoretic Deposition (EPD) method, a technique known for its simplicity and cost-effectiveness. Voltage variations of 20V, 30V, and 40V were applied to both acetone and methanol suspensions for a duration of 20 minutes. The morphology of the Hydroxyapatite/Multiwalled Carbon Nanotube (HA/MWCNT) coatings was meticulously characterized using Scanning Electron Microscopy (SEM). Corrosion resistance was evaluated through Potentiodynamic Polarization (PDP) and Electrochemical Impedance Spectroscopy (EIS) techniques. Remarkably, at 30V, a homogeneous and crack-free coating was achieved, demonstrating superior corrosion resistance. This was further corroborated by the resistance values of 23.891 Ω and 114.990 Ω for the acetone and methanol samples, respectively. Additionally, the corrosion rates of 0.075 (mmpy) and 0.0004 (mmpy) for the acetone and methanol samples further emphasized the superiority of methanol as a suspension medium. These findings unequivocally establish methanol as the optimal choice for achieving superior deposition quality and corrosion resistance in the context of the EPD method for stainless steel type 316L-based dental implants. [ABSTRACT FROM AUTHOR]

Copyright of Turkish Journal of Obstetrics & Gynecology is the property of Galenos Yayinevi Tic. LTD. STI 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.)

Background: Recent studies suggest that bacteria may play a role in triggering rheumatoid arthritis. Considering the recent reports on viable but non-culturable cells (VBNC) in the synovial fluid and blood of patients, the role of bacteria has become more prominent. Objective:The present study aimed to use general and specific primers to detect microbiome in the blood of patients with rheumatoid arthritis. Methods: A general primer, called 16S rRNA was used in the present study to detect a wide range of bacteria in the blood of patients with rheumatoid arthritis. Specific primers were designed and used such as nuc and rfbE to trace Staphylococcus aureus and Escherichia coli in patients' blood. Examining 102 blood samples and performing the genomic extraction separately for each sample, PCR was then performed using general and specific primers, and the results were sequenced. The findings were analyzed using descriptive statistics. Results: According to the results, in 102 blood samples of rheumatoid arthritis patients, who were negative for bacteriological culture, there were 74 cases (72.54%) of 16S rRNA gene, and 54 cases (52.9%) of Staphylococcus aureus nuc gene, respectively. Moreover, a specific gene, rfbE, was traced in 12 samples. Conclusion: The findings of the present study indicated the presence of microbiome in the blood of patients with rheumatoid arthritis. Because the blast sequencing results of the PCR product showed a wide range of bacterial genomes. These findings may potentially improve the management of rheumatoid arthritis diagnosis and treatment. [ABSTRACT FROM AUTHOR]

Background: Electrotransfection is based on application of high-voltage pulses that transiently increase membrane permeability, which enables delivery of DNA and RNA in vitro and in vivo. Its advantage in applications such as gene therapy and vaccination is that it does not use viral vectors. Skeletal muscles are among the most commonly used target tissues. While siRNA delivery into undifferentiated myoblasts is very efficient, electrotransfection of siRNA into differentiated myotubes presents a challenge. Our aim was to develop efficient protocol for electroporation-based siRNA delivery in cultured primary human myotubes and to identify crucial mechanisms and parameters that would enable faster optimization of electrotransfection in various cell lines. Results: We established optimal electroporation parameters for efficient siRNA delivery in cultured myotubes and achieved efficient knock-down of HIF-1α while preserving cells viability. The results show that electropermeabilization is a crucial step for siRNA electrotransfection in myotubes. Decrease in viability was observed for higher electric energy of the pulses, conversely lower pulse energy enabled higher electrotransfection silencing yield. Experimental data together with the theoretical analysis demonstrate that siRNA electrotransfer is a complex process where electropermeabilization, electrophoresis, siRNA translocation, and viability are all functions of pulsing parameters. However, despite this complexity, we demonstrated that pulse parameters for efficient delivery of small molecule such as PI, can be used as a starting point for optimization of electroporation parameters for siRNA delivery into cells in vitro if viability is preserved. Conclusions: The optimized experimental protocol provides the basis for application of electrotransfer for silencing of various target genes in cultured human myotubes and more broadly for electrotransfection of various primary cell and cell lines. Together with the theoretical analysis our data offer new insights into mechanisms that underlie electroporation-based delivery of short RNA molecules, which can aid to faster optimisation of the pulse parameters in vitro and in vivo. [ABSTRACT FROM AUTHOR]

Polysaccharide-based systems have very good emulsifying and stabilizing properties, and starch plays a leading role. Their modifications should add new quality features to the product to such an extent that preserves the structure-forming properties of native starch. The aim of this manuscript was to examine the physicochemical characteristics of the combinations of starch with phospholipids or lysozymes and determine the effect of starch modification (surface hydrophobization or biological additives) and preparation temperature (before and after gelatinization). Changes in electrokinetic potential (zeta), effective diameter, and size distribution as a function of time were analyzed using the dynamic light scattering and microelectrophoresis techniques. The wettability of starch-coated glass plates before and after modification was checked by the advancing and receding contact angle measurements, as well as the angle hysteresis, using the settle drop method as a complement to profilometry and FTIR. It can be generalized that starch dispersions are more stable than analogous n-alkane/starch emulsions at room and physiological temperatures. On the other hand, the contact angle hysteresis values usually decrease with temperature increase, pointing to a more homogeneous surface, and the hydrophobization effect decreases vs. the thickness of the substrate. Surface hydrophobization of starch carried out using an n-alkane film does not change its bulk properties and leads to improvement of its mechanical and functional properties. The obtained specific starch-based hybrid systems, characterized in detail by switchable wettability, give the possibility to determine the energetic state of the starch surface and understand the strength and specificity of interactions with substances of different polarities in biological processes and their applicability for multidirectional use. [ABSTRACT FROM AUTHOR]

The formation of a rigid, tough interface based on a nacre-like structure in carbon fiber (CF) composites is a promising way to eliminate low delamination resistance. An effective method of coating CFs is electrophoretic deposition (EPD), which, in the case of dissimilar components like graphene oxide (GO) and polymeric glue, usually requires chemical bonding/strong interactions. In this work, we focus on chitin nanocrystals (ChNCs), leading to an excellent mechanical performance of artificial nacre, where favorable interactions and bonding with GO are controlled by degrees of deacetylation (5, 15, and 30%). We prepared coatings based on GO/ChNC adducts with 95/5, 90/10, 50/50, and 25/75 ratios using optimized EPD conditions (pH, concentration, voltage, and time). The prepared materials were characterized using FTIR, TEM, XPS, SEM, DLS, and XRD. SEM evaluation indicates the formation of a homogeneous interlayer, which has a fair potential for chemical bonding with the epoxy matrix. Short-beam testing of epoxy matrix composites indicates that the coating does not decrease stiffness and has a relatively low dependence on composition. Therefore, all coatings are promising for a detailed study of delamination resistance using laminate samples. Moreover, facile EPD from the water solution/suspension has a fair potential for industrial applications. [ABSTRACT FROM AUTHOR]

Copyright of Baghdad Science Journal is the property of Republic of Iraq Ministry of Higher Education & Scientific Research (MOHESR) 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.)

Copper-doped ZnO nanoparticles with the formula Zn1−x(Cu)O, where x = 0.0, 0.03, 0.05, and 0.07 were produced using the co-precipitation process. Physical, chemical, and structural properties were properly examined. Powdered X-ray diffraction (P-XRD) patterns revealed the formation of hexagonal wurtzite crystal structure in all samples, through atomic substitutional incorporation in the Cu-doped ZnO lattice. The presence of Cu ions and their dissolution in the host ZnO crystal structure was supported by FT-IR spectra. HR-TEM images were used to assess the average size, morphology, and shape regularity of the synthesized samples. The form and homogeneity of the ZnO changed when Cu ions were substituted, as evidenced by FE-SEM/EDX analysis. The presence of copper signals in the Cu-doped samples indicates that the doping was successful. The decrease in zeta potential with an increased copper doping percentage designates that the nanoparticles (NPs) are more stable, which could be attributed to an increase in the ionic strength of the aqueous solution. The synthesized NPs were evaluated for their substantial in vitro antioxidant properties. In addition, the antimicrobial efficacy of the materials was tested against pathogenic microorganisms. Regarding the anti-diabetic activity, the 7Cu ZnO sample showed the highest inhibitory effect on the α-amylase enzyme. No variations were observed in the activities of the acetylcholinesterase enzyme (AChE) and proteinase enzymes with ZnO and samples doped with different concentrations of Cu. Therefore, further studies are recommended to reveal the in-vitro anti-diabetic activity of the studied doped samples. Finally, molecular docking provided valuable insights into the potential binding interactions of Cu-doped ZnO with α-amylase, FabH of E. coli, and Penicillin-binding proteins of S. aureus. These outcomes suggest that the prepared materials may have an inhibitory effect on enzymes and hold promise in the battle against microbial infections and diabetes. [ABSTRACT FROM AUTHOR]

Background: Familial chylomicronemia syndrome (FCS) is a rare monogenic form of severe hypertriglyceridemia, caused by mutations in genes involved in triglyceride metabolism. Herein, we report the case of a Korean family with familial chylomicronemia syndrome caused by compound heterozygous deletions of glycosylphosphatidylinositol-anchored high-density lipoprotein-binding protein 1 (GPIHBP1). Case presentation: A 4-year-old boy was referred for the evaluation of severe hypertriglyceridemia (3734 mg/dL) that was incidentally detected 4 months prior. His elder brother also demonstrated an elevated triglyceride level of 2133 mg/dL at the age of 9. Lipoprotein electrophoresis revealed the presence of chylomicrons, an increase in the proportion of pre-beta lipoproteins, and low serum lipoprotein lipase levels. The patient's parents and first elder brother had stable lipid profiles. For suspected FCS, genetic testing was performed using the next-generation sequencing-based analysis of 31 lipid metabolism-associated genes, which revealed no pathogenic variants. However, copy number variant screening using sequencing depth information suggested large heterozygous deletion encompassing all the coding exons of GPIHBP1. A real-time quantitative polymerase chain reaction was performed to validate the deletion site. The results showed that the siblings had two heterozygous copy number variants consisting of the whole gene and an exon 4 deletion, each inherited from their parents. During the follow-up period of 17 months, the patient did not develop pancreatitis, following dietary intervention. Conclusion: These siblings' case of familial chylomicronemia syndrome caused by rare GPIHBP1 deletions highlight the implementation of copy number variants—beyond next-generation sequencing—as an important consideration in diagnosis. Accurate genetic diagnosis is necessary to establish the etiology of severe hypertriglyceridemia, which increases the risk of pancreatitis. [ABSTRACT FROM AUTHOR]

Background: Glioblastoma (GBM) stands out as the most prevalent primary brain tumor characterized by its high aggressiveness. Numerous therapeutic approaches have been employed, and the utility of combination therapies has been substantiated, particularly in GBM treatment. Cisplatin, an anticancer chemotherapeutic agent, is employed for the management of various malignancies, including GBM; however, it is associated with significant systemic toxicity. In the realm of combination therapy, metformin, a biguanide drug conventionally used as a first-line treatment for type 2 diabetes, has recently emerged as a valuable adjunct in the treatment of a diverse spectrum of tumors. This study aimed to elucidate the impact of metformin on sensitizing the human cerebral GBM cancer cell line (AMGM) to cisplatin chemotherapy by employing the comet assay as a means to assess DNA damage, thereby advocating the potential of metformin as an adjuvant for cisplatin-based therapy. Method: In this experimental study, the AMGM cell line was cultured and subsequently treated with either single-agent cisplatin, metformin, or a combination of both drugs. Cell viability was assessed through growth inhibition calculations. The Chou--Talalay analysis was used to assess the cooperative effect of this drug combination. Furthermore, DNA fragmentation was quantified using the alkaline comet assay technique. Results: The findings demonstrate that metformin significantly potentiates the therapeutic efficacy of cisplatin by synergistically inhibiting the growth of AMGM cells and reducing DNA damage. Conclusion: These results underscore the potential utility of metformin as a valuable adjunct in enhancing the clinical effectiveness of chemotherapy regimens. [ABSTRACT FROM AUTHOR]

The specific interaction between cell surface receptors and corresponding antibodies has driven opportunities for developing targeted cancer therapies using nanoparticle systems. It is challenging to design and develop such targeted nanomedicines using antibody ligands, as the final nanoconjugate's specificity hinges on the cohesive functioning of its components. The multicomponent nature of antibody-conjugated nanoparticles also complicates the characterization process. Regardless of the type of nanoparticle, it is essential to perform physicochemical characterization to establish a solid foundation of knowledge and develop suitable preclinical studies. A meaningful physicochemical evaluation of antibody-conjugated nanoparticles should include determining the quantity and orientation of the antibodies, confirming the antibodies' integrity following attachment, and assessing the immunoreactivity of the obtained nanoconjugates. In this review, the authors describe the various techniques (electrophoresis, spectroscopy, colorimetric assays, immunoassays, etc.) used to analyze the physicochemical properties of nanoparticles functionalized with antibodies and discuss the main results. [ABSTRACT FROM AUTHOR]

Enterocin P has been characterized in class IIa bacteriocins and showed a broad spectrum of antimicrobial activity. In this study, the nucleotide sequence of enterocin P transformed and expressed in Pichia pastoris genomically, and the antimicrobial activity of the extracted protein was evaluated on pathogenic bacteria. The synthetic sequence of Enterocin P was ligated into pPICZαA, an integrated vector, in the EcoRI - XbaI site. The recombinant plasmid was amplified in Escherichia coli strain DH5α and proved for ligation and probably mutation by sequencing. The linearized plasmid was electroporated into Pichia pastoris. The presence of recombinant protein in yeast cell extraction was verified by SDS-PAGE and the antimicrobial activity of the purified protein. Recombinant Enterocin P tested for the inhibition of 8 avian pathogen bacteria via MIC test. Electrophoresis of purified protein and cell culture supernatant showed a protein band of about 5.4 kDa. The mass spectrometry of the sample exhibits an Amino acid sequence that corresponds to enterocin P in Enterococcus faecium with an accession number of OTN91526.1. Recombinant bacteriocin MIC test shows the most inhibition on Salmonella typhimurium and E. coli O157. Recombinant Pichia pastoris exhibited a potential for producing enterocin P in high concentrations; therefore, this antimicrobial peptide can be studied for pharmacological use or industrial applications. [ABSTRACT FROM AUTHOR]

An analytical method for studying DNA degradation by electrophoresis after cell lysis and visualization of DNA fragments with fluorescent dye, comet assay, was used to evaluate the viability of the endothelial layer of human arterial grafts with the aim of identifying the procedure that will least damage the tissue before cryopreservation. Four groups of samples were studied: cryopreserved arterial grafts that were thawed in two different ways, slowly lasting 2 hours or rapidly for approx. 7 minutes. Arterial grafts that were collected as part of multiorgan procurement with minimal warm ischemia time. Cadaveric grafts were taken as part of the autopsy, so they have a more extended period of warm ischemia. The HeadDNA (%) parameter and others commonly used parameters like TailDNA (%). TailMoment, TailLength, OliveMoment, TailMoment to characterize the comet were used to assess viability in this study. The ratio of nondecayed to decayed nuclei was determined from the values found. This ratio for cadaveric grafts was 0.63, for slowly thawed cryopreserved grafts 2.9, for rapidly thawed cryopreserved grafts 1.9, and for multi-organ procurement grafts 0.68. The results of the study confirmed the assumption that the allografts obtained from cadaveric donors are the least suitable. On the other hand, grafts obtained from multiorgan donors are better in terms of viability monitored by comet assay. [ABSTRACT FROM AUTHOR]

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

Sensitive on-site virus detection is a requirement for timely action against the spread of airborne infection since ultra-low in-air viral concentrations can readily infect individuals when inhaled. Here, we consider a fieldable biosensing process that incorporates a fast RNA enrichment step in order to concentrate viral RNA in a small volume prior to RT-qPCR. The enrichment approach uses electrophoresis in an RT-qPCR-compatible buffer, and allows for concentration of RNA by nearly 5-fold within only 10 min. In order to place this performance into context, we analyzed the minimum detectable concentration of a low-cost, fieldable, biosensing process that uses electrostatic precipitation for air sampling, heating for viral RNA extraction, and then RNA enrichment, followed by RT-qPCR. With enrichment, we estimated an in-air concentration of 5,654 genome copies (gc)/m3 with a 100% detection rate and an in-air concentration of 4,221 gc/m3 with a 78.6% detection rate. Given that the concentrations of common viruses, such as influenza and SARS-CoV-2, in several indoor spaces are between 5,800 and 37,000 gc/m3, we conclude that enrichment allows a detection that is sufficiently sensitive for practical applications.

Wool fiber consists of a complex of protein-keratins, which differ in both their physical and chemical properties and functional characteristics. Despite the progress achieved in the study of the structural organization of fibers, the issue of the influence of environmental factors, the nature and level of nutrition, as well as breed characteristics, remains poorly understood. With this in mind, we investigated the ratio of structural proteins in the wool of sheep of different breeds, which are bred in separate zones of Ukraine, in particular, fine-wool, semi-fine-wool, and coarse-wool. Therefore, during the study of the structural proteins of the wool of sheep of different breeds, which differ among themselves in the thinness of the fibers, it was established that by the method of electrophoretic separation, these proteins are clearly divided into three different groups, in particular, these are proteins of intermediate filaments (IF) of I and II types with a molecular weight of 55–65 and 45–55 kDa, respectively, as well as keratin-associated proteins (KAP) with a molecular weight of 25–35 kDa. However, in some breeds, in particular, the Askanian meat-wool of the black-headed type and the Ukrainian Mountain Carpathian type, two more fractions of proteins were found among IF type I, which almost do not differ from each other in terms of percentage. It was established that within the studied groups of sheep, certain interbreed features are observed, so in the group of thin-wooled breeds, the largest percentage falls on IF II (35.9%). The other two factions account for almost the same percentage — 32.1% (IF I) and 31.8% (KAP). An almost similar picture of the ratio of protein fractions was recorded in the group of animals with coarse wool. At the same time, in the group of sheep with semi-fine wool, in particular, the fibers of sheep of the Ascanian meat-wool breed have the highest percentage of KAP and, accordingly, less IF II, and in the fibers of the Ascanian meat-wool black-headed type, respectively, less proteins of IF II and KAP. Therefore, the obtained data clearly indicate breed characteristics of the content and ratios of certain groups of proteins, which can be genetically determined, and therefore serve as appropriate markers both within individual breeds and within breeds with different fiber categories.

Karin Kaymagi is a type of Tulum cheese traditionally produced from sheep milk and ripened in tripe following the addition of cream to the curd. In this study, it was traditionally produced by mixing certain proportions of raw sheep and cow milk (100:0, 75:25, 50:50, 25:75, and 0:100). The samples were allowed to ripen, and analyses were made for chemical, biochemical, electrophoretic, sensory, and textural properties at the 3rd, 30th, 60th, and 90th days of ripening. According to the results obtained, both cheese type and ripening time had significant effects on dry matter, fat, protein, salt, salt in dry matter, % lactic acid, pH, ripening rate, NPN ratio, and PPN ratio (P < 0.05). Only cheese type had a remarkable effect on salt in dry matter and fat in dry matter (P < 0.05). The highest lipolysis value was determined in the samples made of sheep milk only (KK2) (P < 0.05). Electrophoretic analysis showed that αS1-casein and β-casein concentrations decreased until the end of ripening. Sensory analysis denoted that the KK2 sample was appreciated the most based on colour, appearance, structure, taste, and odor scores (P < 0.05).

Extracellular vesicles (EVs) play important roles in cell-to-cell communications and carry high potential as markers targeted in disease diagnosis, prognosis, and therapeutic development. The main obstacles to EV study are their high heterogeneity; low amounts present in samples; and physical similarity to the abundant, interfering matrix components. Multiple rounds of separation and purification are often needed prior to EV characterization and function assessment. Herein, we report the offline coupling of asymmetrical flow field-flow fractionation (AF4) and capillary electrophoresis (CE) for EV analysis. While AF4 provides gentle and fast EV separation by size, CE resolves EVs from contaminants with similar sizes but different surface charges. Employing Western Blotting, ELISA, and SEM, we confirmed that intact EVs were eluted within a stable time window under the optimal AF4 and CE conditions. We also proved that EVs could be resolved from free proteins and high-density lipoproteins by AF4 and be further separated from the low-density lipoproteins co-eluted in AF4. The effectiveness of the coupled AF4-CE system in EV analysis was demonstrated by monitoring the changes in EV secretion from cells and by direct injection of human serum and detection of serum EVs. We believe that coupling AF4 and CE can provide rapid EV quantification in biological samples with much reduced matrix interference and be valuable for the study of total EVs and EV subpopulations produced by cells or present in clinical samples.

Capillary electrophoresis (CE) is a powerful and sensitive tool for speciation analysis when combined with inductively coupled plasma mass spectrometry (ICP-MS); however, the performance of this technique can be limited by the nature of pneumatic nebulizers. This study compares two commercially available pneumatic nebulizers to a newly introduced vibrating capillary nebulizer (VCN) for on-line coupling of CE with ICP-MS. The VCN is a low-cost, non-pneumatic nebulizer that is based on the design of capillary vibrating sharp-edge spray ionization. As a piezoelectrically driven nebulization source, the VCN creates an aerosol independent of gas flows and does not produce a low-pressure region at the nebulizer orifice. To compare the systems, we performed replicate analyses of sulfate in river water with each nebulizer and the same CE and ICP-MS instruments and determined the figures of merit of each setup. With the CE-VCN-ICP-MS setup, we achieved around 2–4 times lower sensitivity compared to the commercial setups. However, the VCN-based setup provided lower noise levels and better linear correlation from the analysis of calibration standards, which resulted in indistinguishable LOD and LOQ values from the in-house-built VCN-based and commercial setups for CE-ICP-MS analysis. The VCN is found to have the highest baseline stability with a standard deviation of 3500 cts s−1, corresponding to an RSD of 2.7%. High reproducibility is found with the VCN with a peak area RSD of 4.1% between 3 replicate measurements. [ABSTRACT FROM AUTHOR]

Artemisinin is primarily synthesized and stored in the subepidermal space of the glandular trichomes of Artemisia annua. The augmentation of trichome density has been demonstrated to enhance artemisinin yield. However, existing literature lacks insights into the correlation between the stratum corneum and trichomes. This study aims to unravel the involvement of TrichomeLess Regulator 3 (TLR3), which encodes the transcription factor, in artemisinin biosynthesis and its potential association with the stratum corneum. TLR3 was identified as a candidate gene through transcriptome analysis. The role of TLR3 in trichome development and morphology was investigated using yeast two-hybrid, pull-down analysis, and RNA electrophoresis mobility assay. Our research revealed that TLR3 negatively regulates trichome development. It modulates the morphology of Arabidopsis thaliana trichomes by inhibiting branching and inducing the formation of abnormal trichomes in Artemisia annua. Overexpression of the TLR3 gene disrupts the arrangement of the stratum corneum and reduces artemisinin content. Simultaneously, TLR3 possesses the capacity to regulate stratum corneum development and trichome follicle morphology by interacting with TRICHOME AND ARTEMISININ REGULATOR 1, and CycTL. Consequently, our findings underscore the pivotal role of TLR3 in the development of glandular trichomes and stratum corneum biosynthesis, thereby influencing the morphology of Artemisia annua trichomes. [ABSTRACT FROM AUTHOR]

Diffusiophoresis is the isothermal migration of a colloidal particle through a liquid caused by a cosolute concentration gradient. Although diffusiophoresis was originally introduced using hydrodynamics, it can also be described by employing the framework of multicomponent diffusion. This not only enables the extraction of diffusiophoresis coefficients from measured multicomponent-diffusion coefficients but also their theoretical interpretation using fundamental thermodynamic and transport parameters. This review discusses the connection of diffusiophoresis with the 2 × 2 diffusion-coefficient matrix of ternary liquid mixtures. Specifically, diffusiophoresis is linked to the cross-term diffusion coefficient characterizing diffusion of colloidal particles due to cosolute concentration gradient. The other cross-term, which describes cosolute diffusion due to the concentration gradient of colloidal particles, is denoted as osmotic diffusion. Representative experimental results on diffusiophoresis and osmotic diffusion for polyethylene glycol and lysozyme in the presence of aqueous salts and osmolytes are described. These data were extracted from ternary diffusion coefficients measured using precision Rayleigh interferometry at 25 °C. The preferential-hydration and electrophoretic mechanisms responsible for diffusiophoresis are examined. The connection of diffusiophoresis and osmotic diffusion to preferential-interaction coefficients, Onsager reciprocal relations, Donnan equilibrium and Nernst–Planck equations are also discussed. [ABSTRACT FROM AUTHOR]

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 <1 µm) at four different values of average electric field 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. [ABSTRACT FROM AUTHOR]

Background: M’ proteins or paraproteins refer to immunoglobulins that are produced by clonal plasma cells and are a characteristic feature of monoclonal gammopathies. Routine electrophoresis on agarose gel and immunofixation can be used to detect immunoglobulin paraprotein (M-protein). We aimed to evaluate the performance of agarose gel electrophoresis alone and in combination with immunofixation for detecting serum M-proteins. Methods: One hundred and twenty-three patients suspected of paraproteinemia were evaluated. Routine serum protein electrophoresis (SPE) and immunofixation electrophoresis (IFE) protocols were performed. Data from SPE, and SPE-IFE (gel images and electrophoretograms) were collected and reviewed. Results: 21% cases were confirmed using the SPE-IFE combination, and among them, 33% had positive light chain (λ) only on IFE. Similarly, nine cases with biclonal gammopathy on SPE were characterized by IFE. Conclusion: IFE can be a confirmatory test in cases where SPE results are not reliable and it can be a complementary test when characterization of the M protein detected on SPE is required. [ABSTRACT FROM AUTHOR]

Statement of the Problem: Endodontic efforts are focused on eliminating intracanal pathogens. Applying intracanal medicament for infected teeth is beneficial for achieving better antibacterial effects in endodontic treatments. Different intracanal medicaments should be assessed and compared for this purpose. Purpose: The aim of this study was to assess the antibacterial efficacy of Citrullus colocynthis seed extract comparing to Ca(OH)2 on teeth contaminated with Enterococcus faecalis. Materials and Method: In this in vitro study, a novel strain of Enterococcus faecalis (Enterococcus spp. ATCC 19433) yielded from the root canal treated tooth with persistent apical periodontitis. The canals of 78 human single-rooted extracted teeth were contaminated with mentioned strain and treated with Citrullus colocynthis essential oil and Ca(OH)2 for 1, 7, and 14 days. To determine the chemical composition of the oils, gas chromatographymass spectrometry (GC–MS) was applied. The percentage reduction from baseline c.f.u./mL values was estimated. Results: Oleic acid, benzoic acid, and gallic acid were the major contents of Citrullus colocynthis essential oil. The c.f.u./mL count decreased considerably as contact duration rose for both medicaments. After 7 days, a statistically significant difference was identified between the medicaments. Citrullus colocynthis showed higher antimicrobial efficacy. However, after 14 days, no substantial difference was found. Conclusion: Citrullus colocynthis essential oil, displayed great antimicrobial efficacy against Enterococcus faecalis higher than Ca(OH)2 over the first week contact period. [ABSTRACT FROM AUTHOR]