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Nitric oxide (NO) is a redox-dependent signaling molecule that plays a crucial role in regulating a wide range of biological processes in plants. It functions by post-translationally modifying proteins, primarily through S-nitrosation. Thioredoxin (Trx), a small and ubiquitous protein with multifunctional properties, plays a pivotal role in the antioxidant defense system. However, the regulatory mechanism governing the response of tomato Trxh (SlTrxh) to excessive nitrate stress remains unknown. In this study, overexpression or silencing of SlTrxh in tomato led to increased or decreased nitrate stress tolerance, respectively. The overexpression of SlTrxh resulted in a reduction in levels of reactive oxygen species (ROS) and an increase in S-nitrosothiol (SNO) contents; conversely, silencing SlTrxh exhibited the opposite trend. The level of S-nitrosated SlTrxh was increased and decreased in SlTrxh overexpression and RNAi plants after nitrate treatment, respectively. SlTrxh was found to be susceptible to S-nitrosation both in vivo and in vitro , with Cysteine 54 potentially being the key site for S-nitrosation. Protein interaction assays revealed that SlTrxh physically interacts with SlGrx9, and this interaction is strengthened by S-nitrosation. Moreover, a combination of yeast one-hybrid (Y1H), electrophoretic mobility shift assay (EMSA), chromatin immunoprecipitation-quantitative PCR (ChIP-qPCR), and transient expression assays confirmed the direct binding of SlMYB86 to the SlTrxh promoter, thereby enhancing its expression. SlMYB86 is located in the nucleus and SlMYB86 overexpressed and knockout tomato lines showed enhanced and decreased nitrate stress tolerance, respectively. Our findings indicate that SlTrxh functions downstream of SlMYB86 and highlight the potential significance of S-nitrosation of SlTrxh in modulating its function under nitrate stress., Competing Interests: We declare the absence of any conflicts of interest., (© The Author(s) 2024. Published by Oxford University Press on behalf of Nanjing Agricultural University.)

Seed germination and dormancy represent critical phases in the life cycle of plants, tightly regulated by endogenous phytochrome levels and environment signals. High temperatures (HT) induce the overaccumulation of reactive oxygen species (ROS) and increase abscisic acid (ABA), thereby inhibiting seed germination. Our previous findings showed that HT induced the burst of reactive nitrogen species (RNS), increasing the S-nitrosylation modification of HFR1, which effectively blocks seed germination. Importantly, stabilizing HFR1 has been shown to significantly mitigate the inhibitory effect of HT on seed germination. In this study, we reported that HT increased the protein abundance of ABI4, a crucial component in ABA signaling, which in turn activates the expression of RbohD while suppressing the expression of VTC2. This leads to the rapid generation of ROS, thereby inhibiting seed germination. Consistently, the seed germination of abi4 mutant showed insensitivity to HT with lower ROS level during seed germination, whereas transgenic lines overexpressing ABI4 showed reduced germination rates accompanied by elevated ROS levels. Furthermore, we noted that HFR1 interacts with ABI4 to sequester its activity under normal conditions. However, HT-induced ROS triggered the degradation of HFR1, consequently releasing ABI4 activity. This activation of ABI4 promotes RbohD expression, culminating in a ROS burst that suppresses seed germination. Thus, our study unveils a novel function for ABI4 in regulating ROS level and seed germination under HT stress. Throughout this process, HFR1 plays a critical role in restraining ABI4 activity to maintain an optimal endogenous ROS level, thereby ensuring seed germination under favorable environmental conditions., (© 2024 Scandinavian Plant Physiology Society.)

Poa pratensis L. (Poaceae) is a valuable grass across the north hemisphere, inhabiting diverse environments with wide altitudinal span, where ubiquitous various kinds of stresses. Phytohormones would be helpful to improve tolerance to abiotic and biotic stresses, but the responses of transcriptome regulation of P. pratensis to exogenous phytohormones application remain unclear. In this study, we explored the alteration of plant physiological responses by the application of phytohormones. Aiming to achieve this knowledge, we got full-length transcriptome data 42.76 Gb, of which 74.9% of transcripts were completed. Then used 27 samples representing four treatments conducted at two time points (1 h and 6 h after application) to generate RNA-seq data. 371 and 907 common DEGs were identified in response to four phytohormones application, respectively, these DEGs were involved in "plant hormone signal transduction", "carbon metabolism" and "plant-pathogen interaction". Finally, P. pratensis basic research can gain valuable information regarding the responses to exogenous application of phytohormones in physiological indicators and transcriptional regulations in order to facilitate the development of new cultivars., (© 2024. The Author(s).)

We report the synthesis and characterization of a new hybrid magnetic composite formed by the enveloping of magnetic iron oxide nanoparticles (γ-NP) with chains of the conductive polymer PEDOT, and its use for the efficient separation of DNA molecules from complex biological samples, allowing the high yield separation of a pure and high-quality DNA fraction. The successful formation of the γ-NP/PEDOT composite was confirmed by Fourier transform infrared spectroscopy, scanning electron microscopy, UV visible spectroscopy (UV-Vis), and magnetic hysteresis loop measurements. The nanocomposites showed an excellent capacity of DNA adsorption (Q e  ∼ 248 mg/g) in a model system consisting of salmon sperm DNA. When the γ-NP/PEDOT was used in protocols to extract the DNA from complex samples, the corresponding yield was in the range of 6.4 μg (blood) and 7.3 μg (bacteria), as evaluated quality by UV-Vis, PCR analysis, and electrophoresis assays. We also established that the captured DNA does not need to be detached from the nanocomposite for use as seeding material in PCR amplification experiments. These results and the simplicity of the protocols indicate that the γ-NP/PEDOT composite is a promising DNA absorbent, being competitive with the commercially available magnetic purification kits., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

We report the preparation of poly(3,4-ethylene dioxythiophene) (PEDOT)-modified polyvinylidene fluoride electrospun fibers and their use as a novel adsorbent material for the removal of the anionic dye Methyl Orange (MO) from aqueous media. This novel adsorbent material can be used to selectively remove MO on a wide pH range (3.0-10.0), with a maximum capacity of 143.8 mg/g at pH 3.0. When used in a recirculating filtration system, the maximum absorption capacity was reached in a shorter time (20 min) than that observed for batch mode experiments (360 min). Based on the analyses of the kinetics and adsorption isotherm data, one can conclude that the predominant mechanism of interaction between the membrane and the dissolved dye molecules is electrostatic. Besides, considering the estimated values for the Gibbs energy, and entropy and enthalpy changes, it was established that the adsorption process is spontaneous and occurs in an endothermic manner. The good mechanical and environmental stability of these membranes allowed their use in at least 20 consecutive adsorption/desorption cycles, without significant loss of their characteristics. We suggest that the physical-chemical characteristics of PEDOT make these hybrid mats a promising adsorbent material for use in water remediation protocols and effluent treatment systems., (Copyright © 2020. Published by Elsevier B.V.)

Phosphate ions perform a variety of functions in metabolic processes and are essential for all living organisms. The determination of the concentration of phosphate ions is useful in clinical diagnosis of various diseases as an inadequate phosphate level could lead to many health problems. In the search for a cost-effective method of fast monitoring, we investigated the use of cobalt ferrite nanoparticles (CoFeNPs) in the selective recognition of phosphate ions dissolved in aqueous media and more complex samples, such as human blood serum. We prepared these NPs by a chemical coprecipitation route and subjected them to annealing at 600 °C for 1 h. The successful formation of the NPs was confirmed by Fourier transform infrared spectroscopy, X-ray diffraction, transmission electron microscopy, and hysteresis loop measurements. The NPs exhibited a ferrimagnetic behavior, a spinel-type crystalline structure, and hexagonal shape in the nanoscale range. We demonstrated that CoFeNPs containing immobilized fluorescent-labeled single-chain DNA (ssDNA*) probes can be applied for the fast selective detection of phosphate ions dissolved in a liquid medium. We have explored the fact that phosphate groups can displace ssDNA* probes attached to the nanoparticles, therefore causing a perceptible change in the fluorescence signal of the supernatant liquid. This detection method has been tested for the sensing of phosphate ions present both in aqueous solutions and in biological samples, with excellent selectivity and a low limit of detection (∼1.75 nM).

When fluorophores attach to nanostructured films of intrinsically conducting polymers (ICPs), a quenching of their fluorescence may occur. We have exploited these characteristics for the development of polymeric films that can be used in a simple and efficient molecular diagnosis protocol based on the selective detection of nucleic acids. Our procedure rests on the fact that the fluorescence of 6-carboxyfluorescein-labeled single-stranded DNA (FAM-ssDNA) probes is quenched upon their immobilization on nanostructured ICP - polypyrrole (PPY) and polyaniline (PANI) - films deposited on polyethylene terephthalate (PET) substrates. Hybridization occurs whenever a sample with the complementary sequence is brought in contact with the immobilized probe, with the newly formed ds-DNA chains detaching from the flexible polymeric film and causing the restoration of the fluorescence. This sensing system exhibits a low background signal that depends on both the thickness and hydrophobicity of the films. As a model system, we used a FAM-ssDNA probe specific for the Leishmania infantum parasite. The results confirm this procedure as a simple, fast and highly sensitive scheme for the recognition of the target DNA, with a detection limit of the 1.1 nM and 1.3 nM for the PPY/PET and PANI/PET films, respectively. In addition, this biosensor has excellent stability and exhibits a good and reproducible performance even when used for the direct detection of ssDNA in relatively complex biological samples., (Copyright © 2018. Published by Elsevier B.V.)

The development of systems for the controlled release of drugs is important because they allow the control of drug absorption and tissue distribution and also can reduce local toxicity. This study aimed to assemble and characterize two types of release systems, consisting of layer-by-layer films obtained from poly(allylamine) hydrochloride with chlorophyll (PAH/CHL films) or chlorophyll incorporated into dipalmitoylphosphatidylcholine liposomes (PAH/Lip+CHL films). For these systems, the molecular aggregation, growth process, thermally stimulated desorption, wettability, and controlling release of CHL was studied by using UV-vis spectroscopy and wetting contact angle analysis. In addition, experiments of photodynamic inactivation using PAH/CHL or PAH/Lip+CHL films with a 633-nm laser light were performed and the susceptibility of Candida albicans (C. albicans) to this approach was examined. Fluorescence and atomic force microscopies were used to investigate the surface morphology after the application of the photoinactivation procedure. A redshift of the UV-vis spectrum associated to films when compared with the spectrum of the CHL solution indicated a molecular aggregation of CHL molecules in the films. The film growth process was determined by a nucleation and a growth of spheroids or rods for either PAH/Lip+CHL or PAH/CHL films, respectively. Thermally activated desorption experiments indicated that interactions between CHL and PAH (126kJ/mol) in PAH/CHL or between Lip+CHL and PAH (140kJ/mol) in PAH/Lip+CHL films may be governed by electrostatic interactions. The wettability of PAH/Lip+CHL films was larger than that for PAH/CHL films, which can be attributed to hydrophilic groups on the surface of the DPPC liposomes. Release experiments revealed that free CHL in PAH/CHL films was released more slowly than its partner incorporated into liposomes. After the photodynamic inactivation, results of survival fraction and fluorescence microscopy revealed that C. albicans presented similar susceptibility for the two kinds of films. AFM supported the fluorescence one suggesting that cell death of C. albicans may occur due to damages to its cell wall by C. albicans., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Interactions between proteins and drugs, which can lead to formation of stable drug-protein complexes, have important implications on several processes related to human health. These interactions can affect, for instance, free concentration, biological activity, and metabolism of the drugs in the blood stream. Here, we report on the UV-Visible spectroscopic investigation on the interaction of bovine serum albumin (BSA) with chlorophyll (Chl) in aqueous solution under physiological conditions. Binding constants at different temperatures--obtained by using the Benesi-Hildebrand equation--were found to be of the same order of magnitude (~10(4)M(-1)) indicating low affinity of Chl with BSA. We have found a hyperchromism, which suggested an interaction between BSA and Chl occurring through conformational changes of BSA caused by exposition of tryptophan to solvent. Films from BSA and Chl obtained at different Chl concentrations showed fractal structures, which were characterized by fractal dimension calculated from microscopic image analysis.

Introduction: Allogenic hematopoietic stem cell transplant (allo-HSCT) is the treatment of choice for several hematological diseases. Although rare, patients could present nephrotic syndrome as a clinical feature of chronic graft-versus-host disease (cGVHD). The objective of our study is to screen patients with allo-HSCT to determine who developed a glomerular pathology in the context of cGVHD., Patients and Methods: We studied patients who underwent allo-HSCT treatment in our center between October 1995 and October 2012 and who developed glomerular pathology. cGVHD was defined as a pathology when it appeared after 100 d post-allo-HSCT., Results: Five hundred eighty-three allo-HSCT were performed. The prevalence of cGVHD of the kidney was 1.03%. All patients with cGVHD of the kidney were hosts who received peripheral blood from an identical HLA match donor. GVHD prophylaxis with calcineurin inhibitors plus methotrexate was administered in five cases, and prophylaxis with sirolimus was used in another case. cGVHD of the kidney was seen to appear after the removal of the prophylaxis for GVHD, within 33 ± 11.54 months intervals after allo-HSCT in five patients and in another patient, it appeared despite immunosuppressive therapy being administered. All patients had proteinuria, within 11.82 ± 9.03 g/d ranges. The kidney biopsies revealed membranous glomerulonephritis (four patients), focal segmental glomerulonephritis (one patient) and lupus nephropathy class III (one patient). It seems, immunosuppressive therapy achieved complete remission, within the first year of treatment in four patients. Although in three of them, the proteinuria recurred when we tried to remove the therapy; two patients have recently started treatment, being in partial remission now., Conclusions: cGVHD of the kidney is a rare complication after allo-HSCT, related with the removal of the immunosuppression. Monitoring proteinuria in these patients may be useful. In our patients, a complete remission was achieved; although the removal of the immunosuppression may lead to the appearance of outbreaks. We must reconsider the treatment of glomerular pathology secondary to cGVHD., (© 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

This paper reports on the study of the interactions between ascorbic acid (AA) and bovine serum albumin (BSA) in aqueous solution as well as in films (BSA/AA films) prepared by the layer-by-layer technique. Regarding to solution studies, a hyperchromism (in the range of ultraviolet) was found as a function of AA concentration, which suggested the formation of aggregates from AA and BSA. Binding constant, K, determined for aggregates from BSA and AA was found to be about 10(2) M(-1), which indicated low affinity of AA with BSA. For the BSA/AA films, it was also noted that the AA adsorption process and surface morphological structures depended on AA concentration. By changing the contact time between the AA and BSA, a hypochromism was revealed, which was associated to decrease of accessibility of solvent to tryptophan due to formation of aggregates. Furthermore, different morphological structures of aggregates were observed, which were attributed to the diffusion-limited aggregation. Since most of studies of interactions of drugs and proteins are performed in solution, the analysis of these processes by using films can be very valuable because this kind of system is able to employ several techniques of investigation in solid state.