Polyacrylamide Gel Research Articles

Concise Affinity‐Based Purification of Ligated mRNA for Structure‐Activity Relationship Studies of Nucleosugar Modification Patterns

Position‐specific nucleoside sugar modifications have been shown to improve the translational activity and stability of chemically synthesized mRNA. For pharmaceutical applications of chemically modified mRNA, a rapid purification methodology is imperative to identify the optimal modification pattern. However, while the chemical synthesis of mRNAs can be accomplished by splint ligation of oligonucleotide fragments, the current purification method for ligated mRNAs based on denaturing polyacrylamide gel electrophoresis tends to be time consuming. In this study, we developed a two‐step affinity purification method for rapid sample preparation. In this method, ligated mRNA is captured by oligo dT magnetic beads and streptavidin magnetic beads with 3′‐biotinylated oligo DNA, which are complementary to the 3′‐poly(A) and 5′ terminal sequences of the target mRNA, respectively. Therefore, the target mRNA can be isolated from a complex mixture of splint ligations. Using this method, six sugar‐modified mRNAs were simultaneously purified, and the translational activities of these mRNAs were evaluated immediately after purification. The results demonstrate that this methodology is suitable for the rapid preparation of various chemically synthesized mRNAs to identify their optimal modification patterns.

Whey proteins from camel's milk have higher in vitro wound‐healing effect than whey proteins from cow's milk

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

Genetic variations as predictors of dispositional and dyadic empathy—a couple study

Biological drivers of empathy have been explored in an interdisciplinary manner for decades. Research that merges the psychological and genetic perspectives of empathy has recently gained interest, and more complex designs and analyses are needed. Empathy is a multidimensional construct that might be regarded both dispositionally (as a personality trait) and contextually (experienced and/or expressed in a particular relationship/situation). This study analyzed genetic variations associated with genes encoding oxytocin, arginine vasopressin, and receptors that regulate their secretion as predictors of the empathic dimensions of emotional (empathic concern and personal distress) and cognitive (perspective taking) dyadic factors of partners in heterosexual intimate relationships. Machine learning methods to capture both linear and nonlinear relationships between SNPs, RS1 and RS2 repeat polymorphisms and dimensions of empathy in couples were employed. A total of 442 individuals (221 couples) participated in this study. Empathy was measured by the Polish version of the Interpersonal Reactivity Index and the Interpersonal Reactivity Index for Couples. The MassARRAY® 4 instrument, which combines mass spectrometry with endpoint PCR, was used for genotyping all 14 genetic variations. Microsatellite fragment analysis was performed by denaturing polyacrylamide gel electrophoresis. The results confirmed the significance of certain genetic alterations linked to oxytocin, vasopressin, serotonin and estrogen for dispositional and dyadic empathy (mainly rs1884051, rs6311, RS1, rs4686302, and rs1042778) in couples. The effects were stronger for the prediction of emotional and dyadic empathy than for perspective taking. Separate analyses for women and men indicated different predictive effects of genes for empathy (for example, effects of rs53576 were indicated only in women), which are also experienced and expressed in couples. Different dimensions of empathy should be included when the genetic predictors of empathy are examined.

Insights into larval development and protein biochemical alterations of Agrotis ipsilon (Hufnagel) (Lepidoptera: Noctuidae) following Beauveria bassiana and Solanum lycopersicum treatments

BackgroundThe polyphagous notorious pest, black cutworm, Agrotis ipsilon (Hufnagel) (Lepidoptera: Noctuidae), cause significant production losses due to its distinctive feeding and hiding behavior, making it particularly challenging to control it with conventional methods. Therefore, sustainable agriculture demands more effective and environmentally safe pest control solutions. This study aimed to investigate the toxicity of two insecticide alternatives, the entomopathogenic fungus (EPF) Beauveria bassiana and Solanum lycopersicum extract (Tomato plant crude extract, TPCE), using two bioassay methods: the poisoned bait method and the leaf dipping method. In addition, the impact of these biological tools on larval development and protein profiles was evaluated.ResultsThe bait application of both tested materials exhibited higher toxicity than the leaf dipping method, as indicated by the toxicity index. The LC50 values for B. bassiana were 1.6 × 10⁸ and 1.8 × 10⁶ conidia ml−1 using the leaf dipping method and poisoned baits method, respectively. For TPCE, the LC50 values were 4.35 and 1.51 mg ml−1 for the same methods, respectively. In addition, sublethal concentrations of both materials altered the larval and pupal durations. B. bassiana significantly reduced the concentration of larval hemolymph protein. A maximum of 12 protein bands in the control sample, with molecular weights (Mw) ranging between 35 and 120 kDa, were detected by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE). In B. bassiana-treated larvae, ten bands were detected with Mw ranging from 35 to 120 kDa. At least seven bands were detected in TPCE-treated larvae, with Mw ranging from 35 to 97 kDa.ConclusionsThe findings of this study can be integrated into management programs for A. ipsilon. In addition, the availability of B. bassiana and TPCE in Egypt and their cost-effectiveness as insecticide alternatives support their use in the management programs of this critical pest. These methods are particularly effective when applied in bait form.Graphical

Influence of protein in low paste viscosities of Bambara groundnut flours from heat-treated Bambara groundnut seeds

Heat treatment of Bambara groundnut seeds has been reported to cause low paste viscosities in resulting flours. Structural changes in Bambara groundnut protein, due to heat treatment, causes the protein to encapsulate starch, making it unavailable to paste causing low paste viscosities. In this study, trypsin was used to hydrolyze proteins in the flour and the microstructure analysis confirmed the disappearance of aggregates. Flour microstructure analysis confirmed hydrolysis of protein from previously aggregated status and showed liberated individual starch granules. Following the treatment of flours by trypsin, Confocal laser scanning microscopy did not show a protein signal. Hydrolysing Bambara groundnut proteins significantly increased the flour paste viscosities (P < 0.05). The final viscosities for flours from 20 % moisture-conditioned and infrared heat-treated seeds for 5 min were 733.9 mPa s before protein hydrolysis and 2081.71 mPa s after protein hydrolysis. The gelatinization temperature (81 °C) did not show a significant change following protein hydrolysis. Sodium dodecyl-sulphate polyacrylamide gel electrophoresis band intensity increased indicative of disulfide bonding and protein polymerisation when microwave and infrared heat treatment were combined. There were changes in Bambara groundnut protein secondary structures such as an increase of 57 % in β-sheet along with a 60 % reduction in the α-helix as shown by the Fourier-transform infrared spectroscopy. The changes in secondary structure of Bambara groundnut protein were caused by microwave and infrared heating. Heat treatment of Bambara groundnut seeds is partly responsible for the reduction in paste viscosities of their flours.

Enhanced Stability of Hairpin-Functionalized DNA Tetrahedral Nanostructures for miRNA Detection in Plasma from Ischemic Stroke Patients

The enzyme-free amplification technique using the Hybridization Chain Reaction (HCR) is gaining traction for its efficiency in miRNA analysis. Conventional HCR (C-HCR) with hairpin probes faces challenges due to enzymatic degradation in body fluids, leading to potential false-positive results. This study addresses the critical need for a more reliable method that resists enzymatic breakdown and improves diagnostic accuracy for detecting miRNA related to ischemic stroke. We have developed a novel DNA tetrahedral nanostructures-mediated HCR (DTN-HCR) platform for the precise detection of microRNA-25 (miR-25), a biomarker for ischemic stroke. Incorporating two unique DNA tetrahedral nanostructures with embedded hairpin structures (DTN-HP1 and DTN-HP2), this platform activates upon miR-25 binding, initiating a robust DTN-HCR reaction. This reaction forms extensive DNA tetrahedron clusters that significantly boost the fluorescence signal, enabling detection thresholds as low as 5.4 pM. The method showcases exceptional specificity by distinguishing target miRNA from close analogues and maintains structural integrity against DNase I and fetal bovine serum (FBS), verified through polyacrylamide gel electrophoresis (PAGE). It successfully differentiates ischemic stroke patients from healthy controls by analyzing peripheral blood-derived miRNAs. This study concludes that the DTN-HCR platform substantially enhances the specificity and stability of miRNA detection, marking a significant advancement in non-enzymatic miRNA analysis techniques. With its capability to accurately identify ischemic stroke biomarkers at very low concentrations and its resistance to enzymatic degradation, the DTN-HCR method presents a valuable diagnostic tool for ischemic stroke, potentially improving early detection and monitoring in a clinical environment.

Enhanced characterization and utilization of cowpea protein isolate: rheological and textural properties of heat-induced gels for plant-based egg omelet formulation

This study aimed to evaluate the potential of cowpea isolate (CPI) as an effective plant-based egg substitute by comparing its composition and physicochemical properties with those of commercial isolates, including mung bean, soy, and pea proteins. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis identified a distinctive vicilin-like globulin in CPI that is critical for the unique functional properties of CPI. CPI has a smaller particle size, resulting in superior gelation capacity and water retention, which are essential for egg substitute applications. Thermal analysis revealed the high stability of CPI, which is advantageous for food processing. Notably, the unique heat-induced gelation behavior of CPI, characterized by a smooth transition from solid- to liquid-like states, provided a strength not observed in its commercial counterparts. Microstructural analysis showed that the CPI gel closely resembled the egg gel, underscoring the potential of CPI to mimic the texture of eggs, particularly in omelets.

Biochar coupling induced charge transfer switching of a NiAl2O4/NiCr2O4 photocatalyst to enhance photocatalytic degradation of tetracycline hydrochloride

A series of novel (NiAl2O4 (NAO)/NiCr2O4 (NCO)) (S1)/GL heterojunction photocatalysts, denoting as x%GL/S1 (x = 5, 10, 15 and 20), were prepared by the polyacrylamide gel method combined with low temperature sintering technique. Various characterizations confirm that there is special interfacial contact and charge transfer between NAO, NCO and GL. The 10%GL/S1 heterojunction exhibited the best photocatalytic activity and cyclic stability for the degradation of tetracycline hydrochloride (TC). Compared to NAO, NCO and NAO/NCO, the degradation efficiency of 10%GL/S1 was significantly improved, and the degradation percentage reached 83.2% within 30 min. The activity of 10%GL/S1 was studied in relation to catalyst content, drug concentration, pH value, water quality, and co-existing anion. Trapping experiments and free radical verification experiments confirmed that the holes, hydroxyl radicals and superoxide radicals played crucial roles in the photocatalysis process. Possible pathways for TC degradation were studied by HPLC-MS, and the ecotoxicity of TC and its degradation intermediates was evaluated by T.E.S.T. software. The charge transfer and photocatalytic mechanisms confirm that the introduction of GL enhances the interfacial interaction between the components, promotes the carrier migration efficiency, provides a large number of active sites for 10%GL/S1 heterojunction, and produces active free radicals after redox reaction with photoinduced electrons, effectively improving the degradation efficiency of TC.

A new Rose Bengal glycopolymer: Photosensitization in two stages

Antimicrobial photodynamic therapy is a promising alternative to deal with antimicrobial resistance. However, both the low specificity and low local oxygen molecular concentrations decrease the antimicrobial efficiency limiting its use. An interesting approach to the problem is the use of molecules that can react reversibly with singlet oxygen by the formation of reversible endoperoxides, such as naphthalene, anthracene and pyridone derivatives. Particularly, the use of these molecules with mannosyl derivatives allow the interaction with adhesins presented on pili and fimbriae improving the localization near to bacteria. In this work, we synthesized polymeric nanoparticles able to generate singlet oxygen (under both irradiation and dark conditions) in the vicinity of a center capable of recognizing mannose and oxidize nearby biomolecules. Rose Bengal was used as photosensitizer due to its attractive photophysical properties (vis absorption, high singlet oxygen generation) and biocompatibility. The polymeric nanoparticles were obtained by radical polymerization using polyvinyl alcohol as a template, showing sizes around 300 nm with negative zeta potential by dynamic light scattering. The singlet oxygen generation was monitored following DPBF consumption and showed to be dependent on the amount of pyridone in the feed of polymers. In addition, the release of singlet oxygen was also dependent on pyridone concentration showing a slower rate constant at 40 % pyridone, while for contents of 10 % and 60 % higher rate constants were observed. The specific interaction of glycopolymers with Concanavalin A was demonstrated by successful agglutination assays, but also a low participation of unspecific interactions for polymers without mannosyl derivatives was observed. On the other hand, the oxidation of amino acids of Concanavalin A was monitored by acrylamide gel electrophoresis. Type I and Type II photosensitization were observed with the formation of dimers and fragments with lower molecular weight, while in dark conditions only products with lower molecular weight were observed, result consistent with singlet oxygen released by pyridone endoperoxides.

PRP5 Induction by Infection of Pythium Sp. in Tomato

The Pseudomonas fluorescens isolate FP 7 was found to protect the tomato plants against soil borne fungus, Pythium aphanidermatum. The ability to induce defense proteins viz. -1,3 glucanase, peroxidase (PO), polyphenol oxidase (PPO) and phenylalanine ammonia lyase (PAL) by this P. fluorescens isolate FP 7 against P. aphanidermatum fungus was further studied. Increased activity of PO, PPO and -1,3 glucanase were observed in plants pretreated with FP 7 isolate. Native PAGE (polyacrylamide gel electrophoresis) of PO showed the single isoform in all the treatment including the control and difference is by the increased intensity of the band in the inoculated control and FP 7 treatment in the tomato plant. Isoform analysis of the PPO showed the induction of the PPO in the P. fluorescens treated plants challenged with P. aphanidermatum. -1,3 glucanase in tomato cultivar, Co 3 with and without challenge inoculation of P. aphanidermatum, revealed changes in the isoform pattern after staining the gel with 2,3,5-triphenyl tetrazolium chloride. Moreover, higher accumulation of phenolics was noticed in plants pretreated with P. fluorescens isolate FP 7 challenged with P. aphanidermatum. The relevant function of Thaumatin like defense proteins and secondary metabolites involved in the phenylpropanoid pathway collectively contributing to enhanced resistance is discussed.

The Structural Role of RPN10 in the 26S Proteasome and an RPN2-Binding Residue on RPN13 Are Functionally Important in Arabidopsis

The ubiquitin receptors RPN10 and RPN13 harbor multiple activities including ubiquitin binding; however, solid evidence connecting a particular activity to specific in vivo functions is scarce. Through complementation, the ubiquitin-binding site-truncated Arabidopsis RPN10 (N215) rescued the growth defects of rpn10-2, supporting the idea that the ubiquitin-binding ability of RPN10 is dispensable and N215, which harbors a vWA domain, is fully functional. Instead, a structural role played by RPN10 in the 26S proteasomes is likely vital in vivo. A site-specific variant, RPN10-11A, that likely has a destabilized vWA domain could partially rescue the rpn10-2 growth defects and is not integrated into 26S proteasomes. Native polyacrylamide gel electrophoresis and mass spectrometry with rpn10-2 26S proteasomes showed that the loss of RPN10 reduced the abundance of double-capped proteasomes, induced the integration of specific subunit paralogues, and increased the association of ECM29, a well-known factor critical for quality checkpoints by binding and inhibiting aberrant proteasomes. Extensive Y2H and GST-pulldown analyses identified RPN2-binding residues on RPN13 that overlapped with ubiquitin-binding and UCH2-binding sites in the RPN13 C-terminus (246–254). Interestingly, an analysis of homozygous rpn10-2 segregation in a rpn13-1 background harboring RPN13 variants defective for ubiquitin binding and/or RPN2 binding supports the criticality of the RPN13–RPN2 association in vivo.

Optimization of extraction for efficient recovery of kenaf seed protein isolates: evaluation of physicochemical and techno-functional characteristics.

Kenaf seeds are a rich source of protein; however, finding the best extraction method is crucial to obtaining high-quality protein from these underutilized seeds. This research devised an optimized extraction process for best recovery of kenaf seeds protein using response surface methodology. The key parameters affecting the yield and protein content were optimized, including extraction pH (2-11), seed:water ratio (5:1-50:1), temperature (30-90 °C), and duration (20-360 min). The physicochemical and techno-functional properties of kenaf seed protein isolates (KSPIs) were examined. A maximum protein yield of 12.05 g/100 g with purity level 91.94 g/100 g was obtained using an optimized extraction with pH 11.0, seed:water ratio 50:1, 360 min duration, and temperature 50 °C. The oil and water retention capacities of KSPI were 1.14 mL g-1 and 1.37 mL g-1 respectively. After 30 min at pH 7, KSPIs demonstrated remarkable emulsion capacity (83.12%) and stability (75.63%), along with high foaming capacity (106%) and stability (18.3%). As per high-performance liquid chromatography analysis, arginine, glutamic acid, leucine, phenylalanine, and lysine were the most abundant amino acids detected in KPSIs. The KSPIs' globular protein structure was successfully verified using analytical approaches, including Fourier transform infrared spectroscopy, protein fraction ratios, and differential scanning calorimetry. Sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis revealed that KPSI has a molecular weight distribution ranging from 10 kDa to 50 kDa. The results of this study support the application of the proposed response-surface-methodology-optimized extraction method for efficient recovery of high-quality kenaf seed proteins that meet the necessary physicochemical and techno-functional requirements. © 2024 Society of Chemical Industry.

A comprehensive study of AFM stiffness measurements on inclined surfaces: theoretical, numerical, and experimental evaluation using a Hertz approach

Atomic Force Microscopy (AFM) is a leading nanoscale technique known for its significant advantages in the analysis of soft materials and biological samples. Traditional AFM data analysis is often based on the Hertz model, which assumes perpendicular indentation of a planar sample. However, this assumption is not always valid due to the varying geometries of soft materials, whether natural, synthetic or biological. In this study, we present a new theoretical model that incorporates correction coefficients into Hertz’s model to account for cone-like and spherical probes, and to consider local tilt at the probe-sample interface. We validate our model using finite element analysis (FEA) simulations and experimental AFM measurements on tilted polyacrylamide gels. Our results highlight the need to include local tilt at the probe-sample contact to ensure accurate AFM measurements. This represents a step forward in our understanding of the elastic properties at the surface of soft materials in the broadest sense.

A novel m.5906G > a variant in MT-CO1 causes MELAS/Leigh overlap syndrome.

The MELAS/Leigh overlap syndrome manifests with a blend of clinical and radiographic traits from both MELAS and LS. However, the association of MELAS/Leigh overlap syndrome with MT-CO1 gene variants has not been previously reported. In this study, we report a patient diagnosed with MELAS/Leigh overlap syndrome harboring the m.5906G > A variant in MT-CO1, with biochemical evidence supporting the pathogenicity of the variant. The variant m.5906G > A that led to a synonymous variant in the start codon of MT-CO1 was filtered as the candidate disease-causing variant of the patient. Patient-derived fibroblasts were used to generate a series of monoclonal cells carrying different m.5906G > A variant loads for further functional assays. The oxygen consumption rate, ATP production, mitochondrial membrane potential and lactate assay indicated an impairment of cellular bioenergetics due to the m.5906G > A variant. Blue native PAGE analysis revealed that the m.5906G > A variant caused a deficiency in the content of mitochondrial oxidative phosphorylation complexes. Furthermore, molecular biology assays performed for the pathogenesis, mtDNA copy number, mtDNA-encoded subunits, and recovery capacity of mtDNA were all deficient due to the m.5906G > A variant, which might be caused by mtDNA replication deficiency. Overall, our findings demonstrated the pathogenicity of m.5906G > A variant and proposed a potential pathogenic mechanism, thereby expanding the genetic spectrum of MELAS/Leigh overlap syndrome.

Purification and characterization of α-galactosidases from Penicillium griseoroseum for efficient soymilk hydrolysis

Soybean utilization is limited by the presence of raffinose oligosaccharides (RFO), which are not digested by humans and cause gastrointestinal discomfort. This study explores the potential of α-galactosidases from Penicillium griseoroseum for RFO hydrolysis in soymilk. Two distinct α-galactosidase enzymes, designated α-Gal1 and α-Gal2, were purified using a combination of ion-exchange chromatography and native polyacrylamide gel electrophoresis. Both enzymes exhibited characteristics of multimeric proteins and displayed similar biochemical properties. Optimal activity was observed at a pH range of 4.5–5.0 and a temperature range of 40–45 °C. Notably, α-Gal1 demonstrated high thermostability with a half-life of 16 h at 40 °C. The α-galactosidases displayed different substrate affinitiesfor the substrates ρ-NP-αGal, o-NP-αGal, rD-raffinose, d-stachyose, and mD-melibiose. The Michaelis-Menten constant (Km) values for α-Gal1 were 1.06, 1.31, 28.74, 19.88, and 4.77 mmol/L, respectively, while those for α-Gal2 were 0.8, 1.26, 30.46, 21.74 and 5.01 mmol/L, respectively. Both α-Gal1 and α-Gal2 were strongly inhibited by metal ions (Ag⁺, Cu2⁺, Fe2⁺, and Hg2⁺) and moderately inhibited by d-melibiose. Importantly, both enzymes efficiently hydrolyzed RFOs, achieving complete d-stachyose elimination from soymilk after a 6-h incubation. These findings propose the promising application of these α-galactosidases in industrial soymilk production, potentially enhancing its nutritional value and alleviating gastrointestinal issues in consumers.

Evaluation of protective efficacy of recombinant Toxoplasma gondii DDX39 protein vaccine against acute and chronic T. gondii infection in mice

Toxoplasma gondii, a pervasive parasite responsible for toxoplasmosis, poses significant health risks to humans and animals. In this study, we investigated the immunogenicity and protective efficacy of the recombinant T. gondii DDX39 protein formulated with ISA201 adjuvant (rTgDDX39) as a candidate vaccine against toxoplasmosis. The full-length of TgDDX39 gene was successfully amplified, cloned into the pET-30a vector, and expressed in BL21 (DE3) competent cells, which was purified and identified as a 57.1 kDa protein via sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Western blot analysis confirmed that rTgDDX39 was specifically recognized by serum from T. gondii-infected mice. Furthermore, immunization of rats with rTgDDX39 generated antiserum that could specifically recognize the native TgDDX39 protein in T. gondii tachyzoite lysates. Immunofluorescence assay revealed that TgDDX39 was primarily located in the nucleus and perinuclear region of tachyzoites. Our vaccination strategy significantly increased T cell proliferation, with CD4+T cells rising by 21.9% and CD8+T cells by 57.8% by the sixth week compared to the adjuvant control group. Additionally, high titers of anti-rTgDDX39 IgG antibodies were detected in vaccinated mice, with a notable induction of IgG1 and IgG2a isotypes, and IgG1/IgG2a > 1 suggests a Th2-biased immune response.Furthermore, in vitro and in vivo assays demonstrated that polyclonal antibodies raised against rTgDDX39 could inhibit the proliferation of T. gondii RH tachyzoites, highlighting the potential of these antibodies to neutralize this parasite effectively. This study provides compelling evidence of the immunogenicity and protective efficacy of rTgDDX39, supporting its potential as a potential candidate vaccine against toxoplasmosis. The protective efficacy of the vaccine was evaluated in mice challenged with acute (RH) and chronic (PRU) strains of T. gondii, showing a survival time extended to 17 days in the acute model, compared to 13.5 and 14 days in the control groups, and a significant 34% reduction in cyst burden in the chronic model. Additionally, the survival rate in the PRU-infected mice increased from 15 to 20% in the control groups to 45% in the vaccinated group. In vitro and in vivo assays demonstrated that polyclonal antibodies raised against rTgDDX39 could inhibit the proliferation of T. gondii RH tachyzoites, highlighting the potential of these antibodies to neutralize the parasite effectively. This study provides compelling evidence of the immunogenicity and protective efficacy of rTgDDX39, supporting its potential as a candidate vaccine against toxoplasmosis.

A novel γ-chain mutation p.Asp318His in a Chinese family with dysfibrinogenemia.

Congenital dysfibrinogenemia is characterized by reduced fibrinogen activity, but normal immunoreactive fibrinogen levels. Here, we present a novel case with an elevated risk of thrombosis. Coagulation assays, gene analysis, in silico tools, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), fibrin polymerization, thrombin generation assay, and electron microscopy scanning were utilized to elucidate the pathogenic mechanism. The proband manifested with a normal immunologic fibrinogen (2.13 g/l) but reduced functional fibrinogen (0.39 g/l). Subsequent genetic analysis unveiled a novel heterozygous mutation, c.1030G>C (p.Asp318His), in the γ-chain D domain of fibrinogen, which was highly conserved in homologous species and led to enhanced thrombin generation capability. The ability of the proband's fibrinogen to polymerize was significantly impaired, with decreased final turbidity. Scanning electron microscopy indicated that the fibers of the proband were thinner than normal, with smaller pores. Thromboelastography (TEG) results demonstrated prolonged K time, decreased angle value, and a normal confidence interval value in the proband. We present a novel case displaying the γAsp318His mutation, which resulted in dysfibrinogenemia.

Suppression of Zinc Dendrite Growth by Enhanced Polyacrylamide Gel Electrolytes in Zinc-Ion Battery.

Aqueous zinc-ion batteries have become a promising energy storage battery due to high theoretical specific capacity, abundant zinc resources and low cost. However, zinc dendrite growth and hydrogen evolution reaction limit their application. This study aims to improve the cycling performance and stability of aqueous zinc-ion batteries by improving the gel electrolyte. Polyacrylamide (PAM) is selected as the base material of the gel electrolyte, which has good stability and safety, but the water retention capacity, Zn2+ migration number, and ionic conductivity of PAM are low, which affects the long-term stability of the battery. In response to these problems, we optimized PAM by chemical cross-linking method, and formed an enhanced PAM gel by adding disodium citrate dihydrate (SC). Experimental results show that the introduction of an appropriate amount of SC in the enhanced PAM gel electrolyte can significantly improve its electrochemical performance. The zinc-ion symmetric battery achieved a stable cycle of more than 2100 hours at a current density of 0.5 mA cm-2, which is mainly attributed to the inhibitory effect of the enhanced PAM gel on zinc dendrite growth and hydrogen evolution reaction. This study provides a new direction for the development and application of flexible zinc-ion batteries.

Effect of green tea or black tea extract on lipid and protein oxidation in Cantonese sausage.

Natural polyphenols offer a safer alternative to synthetic antioxidants in meat products. This study investigated the efficacy of green tea and black tea extracts as natural antioxidants in Cantonese sausages to inhibit lipid and protein oxidation. Sausages were prepared with the addition of different concentrations - 100, 300, and 600 mg kg-1 total polyphenols (TP) - of green tea or black tea extract. Oxidation of the sausages was assessed through thiobarbituric acid reactants, carbonyl content, and thiol content, whereas consumer acceptability was evaluated based on texture, color, and sensory analysis. The tea extracts inhibited malondialdehyde production and reduced the thiobarbituric acid reactive substance value from 23.72 mmol MDA g-1 to less than 1.94 mmol MDA g-1. However, the addition of tea extracts decreased the thiol content and caused the loss of myosin heavy chain and actin bonds in sodium dodecyl sulfate polyacrylamide gel electrophoresis. Although the addition of tea extracts increased the redness and hardness of the sausage, no significant difference in consumer acceptance between the control and treatment groups was observed in the sensory analysis. The tea extract inhibited the oxidation of lipids in Cantonese sausage. There was no negative effect on the sensory characteristics of sausages. The use of tea extracts as natural antioxidants in Cantonese sausage is therefore feasible. © 2024 Society of Chemical Industry.

Producción de esterasa por Fusarium culmorum crecido en presencia de diferentes concentraciones di(2-etilhexil) ftalato y de ion calcio en fermentación sumergida

Phthalates are plastic additives used as plasticizers in the manufacture of flexible plastic materials. These compounds, such as di(2-ethylhexyl) phthalate (DEHP) are toxic substances to the environment and human health, and have been detected as environmental pollutants. In this study, the activity of esterase produced by Fusarium culmorum grown in media supplemented with different concentrations of DEHP (1.5, 3 and 4 g/L) and calcium ion (0.1 and 0.5 g/L) in submerged fermentation was characterized by biochemical tests and polyacrylamide gel electrophoresis. The greatest esterase activity was observed in cultures grown in media added with 3 g of DEHP/L and 0.5 g of calcium ion/L, followed by those cultures grown in media containing 1.5 g of DEHP/L. Whereas cultures grown on 4 g of DEHP/L had the lowest esterase activity. In general, cultures added with 0.5 g of calcium ion/L had the maximum enzymatic activity and the highest enzymatic productivity in the media tested. High concentration of DEHP were used as the sole source of carbon and energy by F. culmorum. It is suggested that the calcium ion favored the esterase production during the DEHP biodegradation. However, further studies are needed to understand the relationship between calcium ion, fungal growth, and enzyme production to provide the appropriate concentration of this ion in the culture medium.