Soluble Protein Research Articles

Characterization of a missense variant in COG5 in a Tunisian patient with COG5-CDG syndrome and insights into the effect of non-synonymous variants on COG5 protein.

The clinical diagnosis of patients with multisystem involvement including a pronounced neurologic damage is challenging. High-throughput sequencing methods remains crucial to provide an accurate diagnosis. In this study, we reported a Tunisian patient manifesting hypotonia and global developmental delay with visual and skin abnormalities. Exome sequencing was conducted followed by segregation analysis and, subsequently additional investigations. In silico analysis of non-synonymous variants (nsSNPs) described in COG5 in conserved positions was made. Results revealed a homozygous missense variant c.298 C > T (p.Leu100Phe) in the COG5 inherited from both parents. This variant altered both protein solubility and stability, in addition to a putative disruption of the COG5-COG7 interaction. This disruption has been confirmed using patient-derived cells in vitro in a COG5 co-immuno-precipitation, where interaction with binding partner COG7 was abrogated. Hence, we established the COG5-CDG diagnosis. Clinically, the patient shared common features with the already described cases with the report of the ichtyosis as a new manifestation. Conversely, the CADD scoring revealed 19 putatively pathogenic nsSNPs (Minor Allele Frequency MAF < 0.001, CADD > 30), 11 of which had a significant impact on the solubility and/or stability of COG5. These properties seem to be disrupted by six of the seven missense COG5-CDG variants. In conclusion, our study expands the genetic and phenotypic spectrum of COG5-CDG disease and highlight the utility of the next generation sequencing as a powerful tool in accurate diagnosis. Our results shed light on a likely molecular mechanism underlying the pathogenic effect of missense COG5 variants, which is the alteration of COG5 stability and solubility.

Diffusion Tensor Imaging-Along the Perivascular-Space Index Is Associated with Disease Progression in Parkinson's Disease.

The glymphatic clearance pathway is a waste clearance system that allows for removal of soluble proteins such as amyloid β (Aβ) from the brain. Higher Aβ levels are associated with cognitive dysfunction in Parkinson's disease (PD). Diffusion tensor imaging-along the perivascular space (DTI-ALPS) is an imaging measure proposed to indirectly measure glymphatic function. Evaluate differences in DTI-ALPS-index between PD and healthy controls (HC) and characterize relationships between this proposed measure of glymphatic clearance, cognition, and disease severity in PD. PD (n = 32) and HC (n = 23) participants underwent brain imaging to assess DTI-ALPS. PD participants were classified as PD-normal cognition (PD-NC; n = 20) or PD-mild cognitive impairment (PD-MCI; n = 12) based on a Level II comprehensive cognitive assessment. A subgroup of PD participants (n = 21) returned for annual assessments for up to 4 years after baseline. Longitudinal outcomes included changes in performance on the comprehensive cognitive assessment and changes in the Movement Disorders Society Unified Parkinson's Disease Rating Scale (MDS-UPDRS). PD participants had lower DTI-ALPS-index compared to HC. PD participants classified as PD-MCI had significantly lower DTI-ALPS-index compared to PD-NC. Lower DTI-ALPS-index at baseline was associated with longitudinal cognitive decline and worse longitudinal disease severity. Glymphatic clearance, as measured with DTI-ALPS, has potential to serve as a marker of longitudinal disease progression. Interventions targeting glymphatic function should be explored for potential to slow cognitive decline in PD. © 2024 International Parkinson and Movement Disorder Society.

Humic acid and grafting as sustainable agronomic practices for increased growth and secondary metabolism in cucumber subjected to salt stress

Salinity stress poses a significant treat to crop yields and product quality worldwide. Application of a humic acid bio stimulant and grafting onto tolerant rootstocks can both be considered sustainable agronomic practices that can effectively ameliorate the negative effects of salinity stress. This study aimed to assess the above mentioned ameliorative effects of both practices on cucumber plants subjected to saline environments. To attain this goal a factorial experiment was carried out in the form of a completely randomized design with three replications. The three factors considered were (a) three different salinity levels (0, 5, and 10 dS m−1 of NaCl), (b) foliar application of humic acid at three levels (0, 100, and 200 mg L−1), and (c) both grafted and ungrafted plants. Vegetative traits including plant height, fresh and dry weight and number of leaf exhibited a significant decrease under increasing salinity stress. However, the application of humic acid at both levels mitigated these effects compared to control plants. The reduction in relative water content (RWC) of the leaf caused by salinity, was compensated by the application of humic acid and grafting. Thus, the highest RWC (86.65%) was observed in grafting plants with 0 dS m−1 of NaCl and 20 mg L−1 of humic acid. Electrolyte leakage (EL) increased under salinity stress, but the application of humic acid and grafting improved this trait and the lowest amount of EL (26.95%) was in grafting plants with 0 dS m−1 of NaCl and 20 mg L−1 of humic acid. The highest amount of catalase (0.53 mmol H2O2 g−1 fw min−1) and peroxidase (12.290 mmol H2O2 g−1 fw min−1) enzymes were observed in the treatment of 10 dS m−1 of NaCl and 200 mg L−1 humic acid. The highest amount of total phenol (1.99 mg g−1 FW), total flavonoid (0.486 mg g−1 FW), total soluble carbohydrate (30.80 mg g−1 FW), soluble protein (34.56 mg g−1 FW), proline (3.86 µg g−1 FW) was in grafting plants with 0 dS m−1 of NaCl and 200 mg L−1 of humic acid. Phenolic acids and phenylalanine ammonia lyase (PAL) and polyphenol oxidase (PPO) enzymes increased with increasing salinity and humic acid levels. Contrary to humic acid, salt stress increased the sodium (Na+) and chlorine (Cl−) and decreased the amount of potassium (K+) and calcium (Ca2+) in the root and leaf of ungrafted cucumber. However, the application 200 mg L−1 humic acid appeared to mitigate these effects, thereby suggesting a potential role in moderating physiological processes and improving growth of cucumber plants subjected to salinity stress. According to the obtained results, spraying of humic acid (200 mg L−1) and the use of salt resistant rootstocks are recommended to increase tolerance to salt stress in cucumber. These results, for the first time, clearly demonstrated that fig leaf gourd a new highly salt-tolerant rootstock, enhances salt tolerance and improves yield and quality of grafted cucumber plants by reducing sodium transport to the shoot and increasing the amount of compatible osmolytes.

Evaluation of Antimicrobial and Antioxidant Potential of Oxalis corymbosa Extracts.

This work aimed to assess the antioxidant and antimicrobial properties of Oxalis corymbosa extracts. Biochemical analyses were conducted on various plant parts, utilizing enzymatic and non-enzymatic assays. Parameters such as total soluble protein, chlorophyll, and carotenoid contents were also evaluated to elucidate the role of bioactive chemical compounds. The antimicrobial screening of extracts was performed against the bacterial and fungal strains Escherichia coli, Staphylococcus aureus, and Candida albicans, respectively. Results indicated that chlorophyll a, chlorophyll b, total chlorophyll, carotenoid content, anthocyanin content, catalase, peroxidase, and superoxide dismutase were most abundant in the O. corymbosa leaves. Moreover, total ascorbate peroxidase content, total phenolic content, and total flavonoid content were found to be higher in the roots compared to other parts. High-performance liquid chromatography analysis identified chlorogenic acid as the major component, followed by gallic acid, caffeic acid, quercetin, and salicylic acid. Regarding antibacterial potential, each extract exhibited significant activity, with methanolic and ethyl acetate extracts demonstrating the maximum inhibition zone against S. aureus and E. coli, respectively. These findings highlight the substantial antioxidant and antibacterial potential of different parts of O. corymbosa, suggesting their promising applications as ingredients in various nutraceutical products.

Effects of tea infusion on selenium uptake in grapevine

Increased selenium (Se) content in fruits can supply Se in human body, but the effects of teas on the Se uptake in fruit trees are unknown. The effects of infusions of four teas (green, black, dark, and white) on the Se uptake of grapevine were studied to promote the Se uptake in fruit trees in this study. However, only black tea infusion increased the biomass, photosynthetic pigment content, superoxide dismutase (SOD) activity, peroxidase (POD) activity, and soluble protein content of grapevine. Except for white tea infusion, other tea infusions also increased the catalase (CAT) activity of grapevine. Furthermore, the tea infusions increased the activities of adenosine triphosphate sulfurase (ATPS) and adenosine 5’-phosphosulfate reductase (APR), and decreased the activities of serine acetyltransferase (SAT) and selenocysteine methyltransferase (SMT). Only the dark and white tea infusions increased the shoot total Se content by 86.53% and 23.32%, respectively (compared with the control), and also increased the shoot inorganic Se content and shoot organic Se content. Notably, four tea infusions decreased the organic Se proportion and increased the inorganic Se proportion in grapevine. Correlation and grey relational analyses showed that the root total Se content, ATPS activity, and ARP activity were closely associated with the shoot total Se content. The principal component and cluster analyses also showed that the ATPS activity, APR activity, root total Se content, and shoot total Se content were classified into one category. These findings show that black tea infusion can promote grapevine growth, while dark and white tea infusions can promote the Se uptake in grapevine.

Coupled Efficacy of Magneto-Electric Water Irrigation with Foliar Iron Fertilization for Spinach Growth

The lack of trace element iron has become a key factor restricting vegetable yield and quality improvement. To address the low production efficiency of water–fertilizer coupling in agricultural production, we used a combination of experimental research, theoretical analysis, and mathematical modeling to systematically determine the effects of magneto-electric water irrigation with iron application on spinach growth and physiology, its yield and quality, and soil water transport characteristics. Compared with conventional water irrigation, under magneto-electric water irrigation, the water volume and applied iron concentration average increased the total accumulation of nitrogen, carbon, and iron in the aboveground part of spinach by 3.71%, 10.16%, and 14.14%, respectively, and the aboveground part of spinach had the highest total accumulation of nitrogen and carbon when irrigation water was 3300 m3 ha−1 and iron fertilizer application at 0.15%. Additionally, magneto-electric water irrigation increased spinach aboveground fresh weight and soluble sugar and protein content by an average of 13.34, 18.26, and 11.61%, respectively. Based on a comprehensive quantitative evaluation and analysis of aboveground nutrient accumulation in spinach, aboveground fresh weight, water use efficiency, and soluble sugar and protein content, we determined the optimal irrigation water and iron fertilizer application for spinach growth.

Short-term high-light intensity and low temperature improve the quality and flavor of lettuce grown in plant factory.

Lettuce holds a prominent position in the year-round supply of vegetables, offering a rich array of health-beneficial substances, such as dietary fiber, phenolic compounds, lactucopicrin and lactucin. As such, its flavor has garnered increasing attention. Balancing the enhancement of beneficial compounds with the reduction of undesirable taste is a key focus of scientific research. To investigate short-term management to improve the nutritional quality and flavor of lettuce, combinations of different light intensities (200, 500 and 800 μm ol m-2 s-1) and temperatures (10 and 22 °C) were applied separately to 'Lollo Rosso' and 'Little Butter Lettuce' for 7 days before harvest. The results obtained showed that increasing light intensity at low temperatures decreased nitrate content and increased soluble sugar, soluble protein, anthocyanin and phenolic compound content. In the case of lettuce flavor, the bitterness-related metabolites such as lactucin and lactucopicrin were reduced with high light intensity at a low temperature of 10 °C. With this combination, the fructose and glucose contents increased, significantly improving lettuce flavor. Higher light intensity combined with low temperature for 7 days before harvest effectively improved the nutritional quality and flavor of lettuce, suggesting its great potential for use in horticultural practices. © 2024 Society of Chemical Industry.

Peptide production through enhanced solid-state fermentation of soybean meal with Bacillus subtilis SBM_1: Fermentation process and product characteristics

This study aimed to assess the impact of additional inoculation of Bacillus subtilis SBM_1 on solid-state fermentation (SSF) of soybean meal (SBM). The peptide yield of fermented SBM increased to 220.93 mg/g under the optimal SSF conditions, marking a 47.74 % rise compared to natural SSF. Throughout enhanced SSF, the physicochemical properties of metabolites were analyzed. The results showed that the pH and total bacterial counts increased in the initial 8 h, along with elevated levels of peptide, soluble protein, and free amino acid (FAA). Protease activities is maintained at a relatively high level (103.37–241.93 U/g), leading to the substantial degradation of trypsin inhibitor (TI) and macromolecular protein. Following enhanced SSF, the TI activity of the fermented SBM decreased by 76.11 %, and small peptides with molecular weight (MW) < 3000 Da dominated the fermented products (71.64 %). Additionally, the enhanced SSF process demonstrated an improvement in nutrient content in the fermented products. Structural analysis of the fermented SBM showed that macromolecular soybean globulin and allergenic proteins were degraded, with the unfolded and loosened protein spatial structures and the loosely packed and dispersed surface microstructure. Biosafety analysis revealed no detectable microbial contaminants, including mold, salmonella, and coliforms, in compliance with national standards.

Analysis of lettuce transcriptome reveals the mechanism of different light/dark cycle in promoting the growth and quality

Light/dark (L/D) cycle plays a crucial role in controlling the production and quality of vegetables. However, the mechanism of L/D cycle on vegetable growth and quality is scarce studied. To investigate the impact of L/D cycle on lettuce growth and quality, we designed three diel scenarios, including 16 hours of light and 8 hours of darkness (L16/D8), 12 hours of light and 6 hours of darkness (L12/D6), and 8 hours of light and 4 hours of darkness (L8/D4). By phenotypic analysis, we found that lettuce grew taller under the L8/D4 scenario than under L16/D8 light cycle scenarios. The physiological indexes showed that the lettuce leaves grown in the L8/D4 scenario exhibited greater enhancements in the levels of soluble protein, soluble sugar, and carotenoid content compared to the other scenarios. By comparing the expression levels under different diel scenarios (L16/D8 vs L12/D6, L16/D8 vs L8/D4, and L12/D6 vs L8/D4), we identified 7,209 differentially expressed genes (DEGs). Additionally, 3 gene modules that were closely related to L/D cycle of lettuce were selected by WGCNA analysis. The eigengenes of three gene modules were enriched in plant hormone signal transduction, sphingolipid metabolism, and nucleocytoplasmic transport pathways. Through network analysis, we identified six hub genes (CIP1, SCL34, ROPGEF1, ACD6, CcmB, and Rps4) in the three gene modules, which were dominant in plant circadian rhythms and greatly affected lettuce growth. qRT-PCR analysis confirmed the diurnal response patterns of the 6 hub genes in different treatments were significant. This study intensively enhanced our comprehension of the L/D cycle in the growth morphology, nutritional quality, and metabolic pathways of lettuce.

The Influence of Milling Granulation on The Proportion of Fermentable and Non-Fermentable Ingredients in Mash Obtained from Mixed Endosperm Wheat

The use of purpose-selected wheat in brewing is limited because it is either not suitable for the application of usual agrotechnical measures or, more often, is not adapted to specific climatic conditions. As unmalted wheat in brewing can cause process problems and problems with the quality composition of mash/wort and consequently beer, by using the appropriate type and variety, but also by adjusting the malting process, these problems can be completely avoided or significantly reduced at the start. Two varieties were selected for the research, which in previous research showed good brewing/malting properties and which have significantly different starting values for the share of total protein (TP) in the grain. The goal of this research was to determine the effect of grain milling granulation on the quality of mash obtained from a hard type of wheat, i.e., the changes in the fermentable and non-fermentable extract ratio during mashing. Varieties with higher and lower initial contents of total protein (TP) in the grain were tested. It was found that regardless of the initial difference in TP concentration, the concentrations of total soluble proteins (TSP) and high molecular protein fraction (HMW N) are similar for varieties with higher and lower initial TP and that the difference in initial TP concentrations is shown as a difference in the concentration of insoluble protein fractions, which are eliminated during fermentation. By increasing the milling granulation in both kinds of wheat, there is a significant increase in the medium molecular protein fraction (MMW N), which is more pronounced in wheat with a lower starting concentration of TP, even though the starting concentrations of TSP in fine (Ø 0.2 mm) and the superfine milling (Ø &lt; 0.2 mm) were similar. A significant drop in fermentability at the superfine granulation of the grain was noted.

Subconjunctival aflibercept inhibits corneal angiogenesis and VEGFR-3+CD11b+ cells.

This study aimed to investigate the effects of subconjunctival injection of aflibercept, a soluble protein decoy for VEGFR-1 and VEGFR-2, on corneal angiogenesis and VEGFR-expressing CD11b+ cells in a mouse model of suture-induced corneal neovascularization. Corneal neovascularization was induced in BALB/c mice by placing three sutures on the cornea. Immediately after surgery, either 200µg aflibercept (5 µL) or an equal volume of phosphate-buffered saline (PBS) was administered into the subconjunctival space. Seven days after later, corneal new vessels were quantified through clinical examination and measurement of the CD31-stained area in corneal flat mounts. The levels of pro-angiogenic and inflammatory markers in the cornea were evaluated using RT-qPCR. The percentages of VEGFR-2+CD11b+ cells and VEGFR-3+CD11b+ cells were analyzed in the cornea, blood, and draining cervical lymph nodes (DLNs) using flow cytometry. Subconjunctival injection of aflibercept significantly reduced the growth of corneal new vessels compared to subconjunctival PBS injection. The mRNA levels of Cd31, vascular growth factors (Vegfc and Angpt1), and pro-angiogenic/inflammatory markers (Tek/Tie2, Mrc1, Mrc2, and Il6) in the cornea were downregulated by subconjunctival aflibercept. Also, the percentage of VEGFR-3+CD11b+ cells in the cornea, blood, and DLNs was decreased by aflibercept, whereas that of VEGFR-2+CD11b+ cells was unaffected. Subconjunctival aflibercept administration inhibits inflammatory angiogenesis in the cornea and reduces the numbers of cornea-infiltrating and circulating VEGFR-3+CD11b+ cells.

Elaboration and Characterization of Novel Kombucha Drinks Based on Truffles (Tuber melanosporum and Tuber aestivum) with Interesting Aromatic and Compositional Profiles

The organoleptic and bioactive properties of truffles place these fungi as interesting materials for use in the of design functional foods based on fruiting bodies outside commercial standards. Moreover, kombucha beverages have become more popular in the Western world, leading to novel drinks using alternative substrates instead of tea leaves. In this work, two truffle species (Tuber melanosporum, TMEL; Tuber aestivum, TAES) and three different symbiotic consortia of bacteria and yeasts (SCOBYs: SC1, SC2, and SC3) were tested. Fermentation (21 days) was monitored in terms of physicochemical (pH, viscosity), biochemical (total carbohydrates, alcohol, soluble proteins, phenolic compounds), and sensory attributes (volatile organic compounds, VOCs). The obtained pH ranges were adequate, alcohol levels were undetectable or very low, and sugar content was lower than in traditional kombuchas or other beverages. In most cases, the usual bottling time could be applied (7–10 days), although longer fermentations are recommended (14 days) to reach higher protein and phenolic compounds contents. Truffle kombuchas produced up to 51 volatile organic compounds (alcohols, acids, esters, ketones, and aldehydes, among others), with TMEL showing a more complex profile than TAES. During the first week, acidic compound production was observed, especially acetic acid. Similar behavior in the VOC profile was reported with different SCOBYs.

Impact of Surfactants on Cumulative Trypsin Activity in Bottom-Up Proteome Analysis.

Trypsin digestion plays a pivotal role in successful bottom-up peptide characterization and quantitation. While denaturants are often incorporated to enhance protein solubility, surfactants are recognized to inhibit enzyme activity. However, several reports have suggested that incorporating surfactants or other solvent additives may enhance digestion and MS detection. Here, we assess the impacts of ionic surfactants on cumulative trypsin activity and subsequently evaluate the total digestion efficiency of a proteome mixture by quantitative MS. Although low surfactant concentrations, such as 0.01% SDS or 0.2% SDC, significantly enhanced the initial trypsin activity (by 14 or 42%, respectively), time course assays revealed accelerated enzyme deactivation, evident by 10- or 40-fold reductions in trypsin activity half-life at these respective surfactant concentrations. Despite enhanced initial tryptic activity, quantitative MS analysis of a common liver proteome extract, digested with various surfactants (0.01 or 0.1% SDS, 0.5% SDC), consistently revealed decreased peptide counts and signal intensity, indicative of a lower digestion efficiency compared to a nonsurfactant control. Furthermore, including detergents for digestion did not improve the detection of membrane proteins, nor hydrophobic peptides. These results stress the importance of assessing cumulative enzyme activity when optimizing the digestion of a proteome mixture, particularly in the presence of denaturants.

Microplastics in soil affect the growth and physiological characteristics of Chinese fir and Phoebe bournei seedlings

Pot experiments were conducted using Chinese fir (Cunninghamia lanceolata (Lamb.) Hook.) and Phoebe bournei (Hemsl.) Yang) to investigate whether soil microplastics adversely affect the nurturing and renewal of plantations. Microplastics composed of polyethylene and polypropylene with a size of 48 μm were used. The treatments included a control group (without microplastics) and groups treated with microplastic concentrations of 1% and 2% (w/w). The effects of microplastics on the growth, photosynthetic pigments in leaves, antioxidant systems, and osmotic regulation substances of the seedlings were analysed by measuring the seedling height, ground-line diameter growth, chlorophyll (chlorophyll a, chlorophyll b, and total chlorophyll) contents, antioxidant enzyme (superoxide dismutase, peroxidase, catalase) activities, and malondialdehyde, soluble sugar, and soluble protein levels. The results indicated that treatment with 1% polyethylene microplastics increased the chlorophyll a, total chlorophyll, and soluble protein contents in the leaves of both types of seedlings while inhibiting superoxide dismutase and peroxidase activities in P. bournei seedlings. Treatment with 2% polyethylene or polypropylene microplastics suppressed the chlorophyll a, chlorophyll b, and total chlorophyll contents; superoxide dismutase, peroxidase, and catalase activities; and soluble sugar and soluble protein levels in the leaves of both types of seedlings, resulting in reduced growth in terms of height and ground-line diameter. The physiological effects of polyethylene microplastics were more evident than those of polypropylene at the same concentration. The results demonstrated that microplastics can affect photosynthesis, the antioxidant system, and osmotic regulation in Chinese fir and P. bournei seedlings, thereby inhibiting their normal growth and development. Exposure to 1% (w/w) microplastics triggered stress responses in seedlings, whereas 2% (w/w) microplastics impeded seedling growth.

Microplastics in soil affect the growth and physiological characteristics of Chinese fir and Phoebe bournei seedlings

Pot experiments were conducted using Chinese fir (Cunninghamia lanceolata (Lamb.) Hook.) and Phoebe bournei (Hemsl.) Yang) to investigate whether soil microplastics adversely affect the nurturing and renewal of plantations. Microplastics composed of polyethylene and polypropylene with a size of 48 μm were used. The treatments included a control group (without microplastics) and groups treated with microplastic concentrations of 1% and 2% (w/w). The effects of microplastics on the growth, photosynthetic pigments in leaves, antioxidant systems, and osmotic regulation substances of the seedlings were analysed by measuring the seedling height, ground-line diameter growth, chlorophyll (chlorophyll a, chlorophyll b, and total chlorophyll) contents, antioxidant enzyme (superoxide dismutase, peroxidase, catalase) activities, and malondialdehyde, soluble sugar, and soluble protein levels. The results indicated that treatment with 1% polyethylene microplastics increased the chlorophyll a, total chlorophyll, and soluble protein contents in the leaves of both types of seedlings while inhibiting superoxide dismutase and peroxidase activities in P. bournei seedlings. Treatment with 2% polyethylene or polypropylene microplastics suppressed the chlorophyll a, chlorophyll b, and total chlorophyll contents; superoxide dismutase, peroxidase, and catalase activities; and soluble sugar and soluble protein levels in the leaves of both types of seedlings, resulting in reduced growth in terms of height and ground-line diameter. The physiological effects of polyethylene microplastics were more evident than those of polypropylene at the same concentration. The results demonstrated that microplastics can affect photosynthesis, the antioxidant system, and osmotic regulation in Chinese fir and P. bournei seedlings, thereby inhibiting their normal growth and development. Exposure to 1% (w/w) microplastics triggered stress responses in seedlings, whereas 2% (w/w) microplastics impeded seedling growth.

Genetically programmed synthetic cells for thermo-responsive protein synthesis and cargo release.

Synthetic cells containing genetic programs and protein expression machinery are increasingly recognized as powerful counterparts to engineered living cells in the context of biotechnology, therapeutics and cellular modelling. So far, genetic regulation of synthetic cell activity has been largely confined to chemical stimuli; to unlock their potential in applied settings, engineering stimuli-responsive synthetic cells under genetic regulation is imperative. Here we report the development of temperature-sensitive synthetic cells that control protein production by exploiting heat-responsive mRNA elements. This is achieved by combining RNA thermometer technology, cell-free protein expression and vesicle-based synthetic cell design to create cell-sized capsules able to initiate synthesis of both soluble proteins and membrane proteins at defined temperatures. We show that the latter allows for temperature-controlled cargo release phenomena with potential implications for biomedicine. Platforms like the one presented here can pave the way for customizable, genetically programmed synthetic cells under thermal control to be used in biotechnology.

Imaging of Existing and Newly Translated Proteins Elucidates Mechanisms of Sarcomere Turnover.

How the sarcomeric complex is continuously turned over in long-living cardiomyocytes is unclear. According to the prevailing model of sarcomere maintenance, sarcomeres are maintained by cytoplasmic soluble protein pools with free recycling between pools and sarcomeres. We imaged and quantified the turnover of expressed and endogenous sarcomeric proteins, including the giant protein titin, in cardiomyocytes in culture and in vivo, at the single cell and at the single sarcomere level using pulse-chase labeling of Halo-tagged proteins with covalent ligands. We disprove the prevailing protein pool model and instead show an ordered mechanism in which only newly translated proteins enter the sarcomeric complex while older ones are removed and degraded. We also show that degradation is independent of protein age and that proteolytic extraction is a rate-limiting step in the turnover. We show that replacement of sarcomeric proteins occurs at a similar rate within cells and across the heart and is slower in adult cells. Our findings establish a unidirectional replacement model for cardiac sarcomeres subunit replacement and identify their turnover principles.

Evaluation of heavy metal accumulation and tolerance in oxalic acid-treated Phragmites australis wetlands for textile effluent remediation

Water contamination with metals poses significant environmental challenges. The occurrence of heavy metals (HMs) prompts modifications in plant structures, emphasizing the necessity of employing focused safeguarding measures. Cadmium (Cd), lead (Pb), and chromium (Cr) emerge as particularly menacing toxins due to their high accumulation potential. Increasing the availability of organic acids is crucial for optimizing toxic metal removal via phytoremediation. This constructed wetland system (CWs) was used to determine how oxalic acid (OA) treatments of textile wastewater (WW) effluents affected morpho-physiological characteristics, antioxidant enzyme activity, oxidative stress, and HM concentrations in Phragmites australis. Multiple treatments, comprising the application of OA at a concentration of 10 mM and WW at different dilutions (25%, 50%, 75%, and 100%), were employed, with three replications of each treatment. WW stress decreased chlorophyll and carotenoid content, and concurrently enhanced HMs adsorption and antioxidant enzyme activities. Furthermore, the application of WW was found to elevate oxidative stress levels, whereas the presence of OA concurrently mitigated this oxidative stress. Similarly, WW negatively affected soil-plant analysis development (SPAD) and the total soluble proteins (SP) in both roots and shoots. Conversely, these parameters showed improvement with OA treatments. P. australis showed the potential to enhance HM accumulation under 100% WW stress. Specifically, there is an increase in root SP ranging from 9% to 39%, an increase in shoot SP from 6% to 91%, and an elevation in SPAD values from 4% to 64% compared to their respective treatments lacking OA inclusion. The OA addition resulted in decreased EL contents in the root and shoot by 10%–19% and 13%–15%, MDA by 9%–14% and 9%–20%, and H2O2 by 14%–21% and 9%–17%, in comparison to the respective treatments without OA. Interestingly, the findings further revealed that the augmentation of OA also contributed to an increased accumulation of Cr, Cd, and Pb. Specifically, at 100% WW with OA (10 mM), the concentrations of Cr, Pb, and Cd in leaves rose by 164%, 447%, and 350%, in stems by 213%, 247%, and 219%, and in roots by 155%, 238%, and 195%, respectively. The chelating agent oxalic acid effectively alleviated plant toxicity induced by toxins. Overall, our findings demonstrate the remarkable tolerance of P. australis to elevated concentrations of WW stress, positioning it as an eco-friendly candidate for industrial effluent remediation. This plant exhibits efficacy in restoring contaminants present in textile effluents, and notably, oxalic acid emerges as a promising agent for the phytoextraction of HMs.

P-290 Isolation and characterization of extracellular vesicles from human endometrial organoids

Abstract Study question Do human endometrial organoids produce extracellular vesicles (EVs)? Summary answer Endometrial organoids release EVs (ORG-EVs) that we successfully and comprehensively characterize according to guidelines. What is known already EVs play a crucial role in cellular communication, garnering increasing attention in physiological and pathological processes. Although the enormous potential of EVs has been extensively demonstrated across a large variety of research fields, the promise held by EVs derived from human endometrial organoids is still largely unexplored. Indeed, nowadays there is limited research available on the presence, role, and function of EVs derived from organoids of female reproductive tissue samples. This underscores the importance of adopting standardized methods for isolating, characterizing, and reporting findings, which is crucial for advancing in this promising and innovative, yet challenging, research field. Study design, size, duration This study involved establishing organoid cultures from endometrial biopsies of healthy women (n = 10) of reproductive age, who underwent ovarian surgery between January and October 2023. ORG-EVs were isolated using differential ultracentrifugation. Characterization of ORG-EVs, including assessment of their biophysical and biochemical properties, adhered to guidelines established by the International Society for Extracellular Vesicles (ISEV). Participants/materials, setting, methods Endometrial specimens from 10 women were employed to establish organoid cultures. ORG-EVs were isolated from spent medium through differential ultracentrifugation. Morphology, size distribution and concentration analysis were performed using Transmission Electron Microscopy (TEM) and Nanoparticle Tracking Analysis (NTA). Western blotting was used to confirm the presence of EVs markers. Main results and the role of chance TEM analysis confirmed the presence of isolated ORG-EVs, elucidating their morphology and structural integrity. NTA revealed that ORG-EVs had an average size of 160.4 ± 6.2 nm (mean ± SD) and a mean concentration of 5.2E+11 ± 2.1E+10 particles/ml (mean ± SD). Western blot analysis further confirmed the vesicular nature of the assessed particles, demonstrating positivity for standard EV markers such as tetraspanins (CD63 and CD9), and soluble proteins (ALIX and TSG101), while excluding the endoplasmic reticulum-resident protein calnexin (negative control). Additionally, ORG-EVs exhibited a positive signal for mucin1 (MUC1), a glycoprotein present on the surface of endometrial epithelial cells, thereby confirming their cellular origin. Limitations, reasons for caution This is a pilot study, characterized by a small sample size requiring larger cohorts to validate results and, at the same time, to undertake deeper exploration of ORG-EVs’ molecular cargo. Emphasizing the critical need for standardized isolation and characterization methods is paramount. Wider implications of the findings Endometrial organoids enhance EVs production, providing a powerful tool for reproductive aspects studies, and improving regenerative medicine and artificial reproductive technology. These initial findings propel further exploration, unveiling of ORG-EVs’ molecular cargo aiming to elucidate their potential role in human female reproductive tissues and mechanism related to embryo implantation process. Trial registration number not applicable

Foliar application of agrochemicals enhance growth, productivity, quality and biochemical traits of soybean

Flower drop, senescence of flowers and inadequate pod filling pose significant challenges in soybean cultivation as rapid nutrient extraction from leaves during the reproductive stage accelerates leaf aging. The present study assessed the effect of foliar application of agrochemicals on growth, productivity, quality and biochemical traits of soybean at two locations during kharif (rainy) season of 2021 and 2022. The experiment comprised of ten treatments of foliar application of agrochemicals [absolute control, water spray, potassium nitrate (KNO3) 1.5%, NPK (19:19:19) 1.5%, boron 0.3%, ammonium molybdate (AM) 0.5%, thiourea 750 parts per million (ppm), urea 2.0%, muriate of potash (MOP) 0.5% and salicylic acid (SA) 100 ppm] at flower and pod initiation stages along with the recommended dose of fertilizer (RDF) in a randomized complete block design replicated thrice. Results revealed that foliar application of boron 0.3%, urea 2.0% and MOP 0.5% at Ludhiana and MOP 0.5%, potassium nitrate 1.5% and urea 2.0% at Ballowal Saunkhri increased growth (plant height and dry matter accumulation), yield attributes (number of pods plant−1, number of filled pods plant−1 and 100-seed weight) and seed yield. These agrochemicals showed positive effects on the biochemical parameters with increased levels of proline and total phenol content as well as improvements in total soluble sugars, starch and protein content. The findings suggest that foliar application of agrochemicals (boron 0.3%, urea 2.0%, MOP 0.5% and potassium nitrate 1.5%) along with RDF can improve soybean growth and seed yield by mitigating the nutrient stress at reproductive stages.