Exploring the dynamics of ISR signaling in maize upon seed priming with plant growth promoting actinobacteria isolated from tea rhizosphere of Darjeeling.

Plant growth-promoting rhizobacteria (PGPR) offer an eco-friendly alternative to agrochemicals for better plant growth and development. Here, we evaluated the plant growth promotion abilities of actinobacteria isolated from the tea (Camellia sinensis) rhizosphere of Darjeeling, India. 16 S rRNA gene ribotyping of 28 isolates demonstrated the presence of nine different culturable actinobacterial genera. Assessment of the in vitro PGP traits revealed that Micrococcus sp. AB420 exhibited the highest level of phosphate solubilization (i.e., 445 ± 2.1 µg/ml), whereas Kocuria sp. AB429 and Brachybacterium sp. AB440 showed the highest level of siderophore (25.8 ± 0.1%) and IAA production (101.4 ± 0.5 µg/ml), respectively. Biopriming of maize seeds with the individual actinobacterial isolate revealed statistically significant growth in the treated plants compared to controls. Among them, treatment with Paenarthrobacter sp. AB416 and Brachybacterium sp. AB439 exhibited the highest shoot and root length. Biopriming has also triggered significant enzymatic and non-enzymatic antioxidative defense reactions in maize seedlings both locally and systematically, providing a critical insight into their possible role in the reduction of reactive oxygen species (ROS) burden. To better understand the role of actinobacterial isolates in the modulation of plant defense, three selected actinobacterial isolates, AB426 (Brevibacterium sp.), AB427 (Streptomyces sp.), and AB440 (Brachybacterium sp.) were employed to evaluate the dynamics of induced systemic resistance (ISR) in maize. The expression profile of five key genes involved in SA and JA pathways revealed that bio-priming with actinobacteria (Brevibacterium sp. AB426 and Brachybacterium sp. AB440) preferably modulates the JA pathway rather than the SA pathway. The infection studies in bio-primed maize plants resulted in a delay in disease progression by the biotrophic pathogen Ustilago maydis in infected maize plants, suggesting the positive efficacy of bio-priming in aiding plants to cope with biotic stress. Conclusively, this study unravels the intrinsic mechanisms of PGPR-mediated ISR dynamics in bio-primed plants, offering a futuristic application of these microorganisms in the agricultural fields as an eco-friendly alternative.

Ger1 is a secreted aspartic acid protease essential for spore germination in Ustilago maydis.

Ustilago maydis expresses a number of proteases during its pathogenic lifecycle. Some of the proteases including both intracellular and extracellular ones have previously been shown to influence the virulence of the pathogen. However, any role of secreted proteases in the sporulation process of U. maydis have not been explored earlier. In this study we have investigated the biological function of one such secreted protease, Ger1 belonging to aspartic protease A1 family. An assessment of the real time expression of ger1 revealed an infection specific expression of the protein especially during late phases of infection. We also evaluated any contribution of the protein in the pathogenicity of the fungus. Our data revealed an involvement of Ger1 in the sporulation and spore germination processes of U. maydis. Ger1 also showed positive influence on the pathogenicity of the fungus and accordingly the ger1 deletion mutant exhibited reduced pathogenicity. The study also demonstrated the protease activity associated with Ger1 to be essential for its biological function. Fluorescence microscopy of maize plants infected with U. maydis cells expressing Ger1-mcherry-HA also revealed that Ger1 is efficiently secreted within maize apoplast.

Defense Surveillance System at the Interface: Response of Rice Towards Rhizoctonia solani During Sheath Blight Infection.

Sheath blight of rice caused by necrotrophic plant pathogen Rhizoctonia solani is one of the most common fungal diseases of rice leading to significant yield loss. Among the defense responses exhibited by the host plants towards fungal infections, those functional within the apoplast contribute significantly. Here, we have studied apoplastic defense response of rice towards R. solani during sheath blight infection. The transcriptome of R. solani-infected rice plants was compared with that of uninfected rice, to identify the set of defense genes that undergo differential expression and code for proteins with a predicted N-terminal signal peptide. Significant changes in the stress-responsive, molecular signal perception, protein modification, and metabolic process pathways represented by a group of differentially expressed genes were observed. Our data also revealed two secreted protease inhibitors from rice that exhibit increased expression during R. solani infection and induce disease resistance when expressed in Nicotiana benthamiana. [Formula: see text] Copyright © 2022 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Ustilago maydis secreted T2 ribonucleases, Nuc1 and Nuc2 scavenge extracellular RNA.

Ustilago maydis genome codes for many secreted ribonucleases. The contribution of two among these belonging to the T2 family (Nuc1 and Nuc2) in the pathogen virulence, has been assessed in this study. The nuc1 and nuc2 deletion mutants showed not only reduced pathogenicity compared to the SG200 WT strain but also exhibited significant delay in the completion of the pathogenic lifecycle. Both the proteins were also tested for their nucleolytic activities towards RNA substrates from maize and yeast. This also yielded valuable insights into the ability of the ribonucleases to utilise extracellular RNA as a nutrient source. Our study therefore established a role of two T2 type secreted ribonucleases of a phytopathogen in the acquisition of nutrient for the first time. This study also provides evidence that maize apoplast contains RNA, which can be utilised as a substrate by both Nuc1 and Nuc2.

Induction of apoptosis-like cell death and clearance of stress-induced intracellular protein aggregates: dual roles for Ustilago maydis metacaspase Mca1.

Metacaspases primarily associate with induction and execution of programmed cell death in protozoa, fungi and plants. In the recent past, several studies have also demonstrated cellular functions of metacaspases other than cell death in different organisms including yeast and protozoa. This study shows similar dual function for the only metacaspase of a biotrophic phytopathogen, Ustilago maydis. In addition to a conventional role in the induction of cell death, Mca1 has been demonstrated to play a key role in maintaining the quality of the cellular proteome. On one hand, Mca1 could be shown to bring about apoptosis-like phenotypic changes in U. maydis on exposure to oxidative stress, on the other hand, the protein was found to regulate cellular protein quality control. U. maydis metacaspase has been found to remain closely associated with the insoluble intracellular protein aggregates, generated during an event of stress exposure to the fungus. The study, therefore, provides direct evidence for a role of U. maydis metacaspase in the clearance of the stress-induced intracellular insoluble protein aggregates. Furthermore, host infection assays with mca1 deletion strain also revealed a role of the protein in the virulence of the fungus.

Metabolic priming by a secreted fungal effector.

Maize smut caused by the fungus Ustilago maydis is a widespread disease characterized by the development of large plant tumours. U. maydis is a biotrophic pathogen that requires living plant tissue for its development and establishes an intimate interaction zone between fungal hyphae and the plant plasma membrane. U. maydis actively suppresses plant defence responses by secreted protein effectors. Its effector repertoire comprises at least 386 genes mostly encoding proteins of unknown function and expressed exclusively during the biotrophic stage. The U. maydis secretome also contains about 150 proteins with probable roles in fungal nutrition, fungal cell wall modification and host penetration as well as proteins unlikely to act in the fungal-host interface like a chorismate mutase. Chorismate mutases are key enzymes of the shikimate pathway and catalyse the conversion of chorismate to prephenate, the precursor for tyrosine and phenylalanine synthesis. Root-knot nematodes inject a secreted chorismate mutase into plant cells likely to affect development. Here we show that the chorismate mutase Cmu1 secreted by U. maydis is a virulence factor. The enzyme is taken up by plant cells, can spread to neighbouring cells and changes the metabolic status of these cells through metabolic priming. Secreted chorismate mutases are found in many plant-associated microbes and might serve as general tools for host manipulation.

Hailey-Hailey Disease Presenting as Lichenoid Plaques on the Thigh.

A 40-year-old woman presented with multiple itchy violaceous papules and plaques on the medial aspects of both thighs. This was accompanied by a burning sensation on friction and increased itching that interfered with her daily activities, particularly in the summer. The family history was noncontributory. Cutaneous examination showed multiple flat-topped hyperkeratotic violaceous papules and plaques, appearing in linear fashion on the medial aspects of the thighs bilaterally (Figure 1). Histopathologic examination showed acanthosis, widespread suprabasal acantholysis resembling a "dilapidated brick wall," without any evidence of dyskeratosis. There was a mild dermal perivascular infiltrate (Figure 2). On the basis of the clinical and histologic findings, she was diagnosed as having localized Hailey-Hailey disease. She was treated conservatively with topical corticosteroids and antimicrobials. She was also advised to lose weight, to avoid friction, and to wear loose-fitting cotton garments. There was marked symptomatic relief, and the lesion showed mild improvement over the next 2 months.

Deciphering the mode of action of a mutant Allium sativum Leaf Agglutinin (mASAL), a potent antifungal protein on Rhizoctonia solani.

BACKGROUND: Mutant Allium sativum leaf agglutinin (mASAL) is a potent, biosafe, antifungal protein that exhibits fungicidal activity against different phytopathogenic fungi, including Rhizoctonia solani. METHODS: The effect of mASAL on the morphology of R.solani was monitored primarily by scanning electron and light microscopic techniques. Besides different fluorescent probes were used for monitoring various intracellular changes associated with mASAL treatment like change in mitochondrial membrane potential (MMP), intracellular accumulation of reactive oxygen species (ROS) and induction of programmed cell death (PCD). In addition ligand blot followed by LC-MS/MS analyses were performed to detect the putative interactors of mASAL. RESULTS: Knowledge on the mode of function for any new protein is a prerequisite for its biotechnological application. Detailed morphological analysis of mASAL treated R. solani hyphae using different microscopic techniques revealed a detrimental effect of mASAL on both the cell wall and the plasma membrane. Moreover, exposure to mASAL caused the loss of mitochondrial membrane potential (MMP) and the subsequent intracellular accumulation of reactive oxygen species (ROS) in the target organism. In conjunction with this observation, evidence of the induction of programmed cell death (PCD) was also noted in the mASAL treated R. solani hyphae. Furthermore, we investigated its interacting partners from R. solani. Using ligand blots followed by liquid chromatography tandem mass spectrometry (LC-MS/MS) analyses, we identified different binding partners including Actin, HSP70, ATPase and 14-3-3 protein. CONCLUSIONS: Taken together, the present study provides insight into the probable mode of action of the antifungal protein, mASAL on R. solani which could be exploited in future biotechnological applications.

Molecular and functional characterisation of a stress responsive cysteine protease, EhCP6 from Entamoeba histolytica.

Entamoeba histolytica cysteine protease 6 (EhCP6) is a stress responsive cysteine protease that is upregulated in response to heat shock and during pathogen invasion of the host tissue. In the present study an attempt has been made to express and purify recombinant EhCP6 in order to gain insights into its biochemical properties. The recombinant and refolded protein has been shown to undergo autoproteolysis in the presence of DTT and SDS to give rise to ∼25kDa mature form. The mature form of the protein was found to exhibit a protease activity that is sensitive to E-64, a specific cysteine protease inhibitor. In silico homology modelling of EhCP6 revealed that the protein exhibits conservation of almost all the major structural features of cathepsin-L like cysteine proteases. Further in vivo studies are needed to decipher the function of the protein in response to different stressed conditions.

Insight into the biochemical and cell biological function of an intrinsically unstructured heat shock protein, Hsp12 of Ustilago maydis.

Small heat shock proteins (sHsps) play diverse roles in the stress response and maintenance of cellular functions. The Ustilago maydis genome codes for few sHsps. Among these, Hsp12 has previously been demonstrated to be involved in the pathogenesis of the fungus by our group. In the present study we further investigated the biological function of the protein in the pathogenic development of U. maydis. Analysis of the primary amino acid sequence of Hsp12 in combination with spectroscopic methods to analyse secondary protein structures revealed an intrinsically disordered nature of the protein. We also carried out detailed analysis on the protein aggregation prevention activity associated with Hsp12. Our data suggest Hsp12 has trehalose-dependent protein aggregation prevention activity. Through assaying the interaction of Hsp12 with lipid membranes in vitro we also showed the ability of U. maydis Hsp12 to induce stability in lipid vesicles. U. maydis hsp12 deletion mutants exhibited defects in the endocytosis process and delayed completion of the pathogenic life cycle. Therefore, U. maydis Hsp12 contributes to the pathogenic development of the fungus through its ability to relieve proteotoxic stress during infection as well as its membrane-stabilizing function.

Carbon Nanomaterial-Based Hydrogels as Scaffolds in Tissue Engineering: A Comprehensive Review.

Carbon-based nanomaterials (CBNs) are a category of nanomaterials with various systems based on combinations of sp2 and sp3 hybridized carbon bonds, morphologies, and functional groups. CBNs can exhibit distinguished properties such as high mechanical strength, chemical stability, high electrical conductivity, and biocompatibility. These desirable physicochemical properties have triggered their uses in many fields, including biomedical applications. In this review, we specifically focus on applying CBNs as scaffolds in tissue engineering, a therapeutic approach whereby CBNs can act for the regeneration or replacement of damaged tissue. Here, an overview of the structures and properties of different CBNs will first be provided. We will then discuss state-of-the-art advancements of CBNs and hydrogels as scaffolds for regenerating various types of human tissues. Finally, a perspective of future potentials and challenges in this field will be presented. Since this is a very rapidly growing field, we expect that this review will promote interdisciplinary efforts in developing effective tissue regeneration scaffolds for clinical applications.

Archaeal Hsp14 drives substrate shuttling between small heat shock proteins and thermosome: insights into a novel substrate transfer pathway.

Heat shock proteins maintain protein homeostasis and facilitate the survival of an organism under stress. Archaeal heat shock machinery usually consists of only sHsps, Hsp70, and Hsp60. Moreover, Hsp70 is absent in thermophilic and hyperthermophilic archaea. In the absence of Hsp70, how aggregating protein substrates are transferred to Hsp60 for refolding remains elusive. Here, we investigated the crosstalk in the heat shock response pathway of thermoacidophilic crenarchaeon Sulfolobus acidocaldarius. In the present study, we biophysically and biochemically characterized one of the small heat shock proteins, Hsp14, of S. acidocaldarius. Moreover, we investigated its ability to interact with Hsp20 and Hsp60 to facilitate the substrate proteins' folding under stress conditions. Like Hsp20, we demonstrated that the dimer is the active form of Hsp14, and it forms an oligomeric storage form at a higher temperature. More importantly, the dynamics of the Hsp14 oligomer are maintained by rapid subunit exchange between the dimeric states, and the rate of subunit exchange increases with increasing temperature. We also tested the ability of Hsp14 to form hetero-oligomers via subunit exchange with Hsp20. We observed hetero-oligomer formation only at higher temperatures (50 °C-70 °C). Furthermore, experiments were performed to investigate the interaction between small heat shock proteins and Hsp60. We demonstrated an enthalpy-driven direct physical interaction between Hsp14 and Hsp60. Our results revealed that Hsp14 could transfer sHsp-captured substrate proteins to Hsp60, which then refolds them back to their active form.

Evaluation of various nephrometric scoring system for nephrolithiasis to predict stone free status after percutaneous nephrolithotomy: A comparative study.

BACKGROUND: The CROES Nephrolithometry nomogram, S.T.O.N.E. Nephrolithometry Score and Guy's stone score were introduced for stratification of kidney stones disease on the basis of quantitative stone burden and its distribution. Till date there has been very limited data on head to head comparison of the existing scoring systems. Comparison and analyses among the scoring system helps in further refinement of these systems along with development of new more effective and broadly acceptable nomogram. OBJECTIVE: Predictability of the stone-free status (SFS) and post-operative complication after PCNL by various scoring systems (The CROES nomogram, S.T.O.N.E. nephrolithometry score and Guy's stone score). MATERIALS AND METHODS: Total 100 adult patients underwent PCNL after considering inclusion and exclusion criteria. All patients underwent Preoperative NCCT scan, investigations of blood (Hb%, PCV, bleeding and coagulation profile, urea, and creatinine), and urine (RE/ME and C/S), Postoperative X ray KUB/NCCT. RESULTS: ROC curves were developed for each scoring system to determine the accuracy to predict stone free status. We found CS had significantly higher AUC than other scoring systems [p-value for CS vs GSS = 0.0091 & CS vs SS = 0.000]. So CS has higher accuracy to predict stone free status. None of the scoring system had shown significantly higher AUC than other scoring system in predicting complication. CONCLUSION: CROES Nephrolithometry nomogram is most accurate to predict preoperative stone-free rate. All scoring systems can equally predict perioperative complications and other variables.

Production of rGO-Based Electrospinning Nanocomposites Incorporated in Recycled PET as an Alternative Dry Electrode.

In this work, Coca-Cola® bottles were reused as a PET polymer (rPET) source to produce electrospun polymeric nanofibers. The nanofibers were electrospun from polymer solutions with different concentrations of reduced graphene oxide (rGO) incorporated for applications in somatosensory electrical stimulation. The rPET/rGO nanofiber mats were characterized by SEM, TEM, Raman, DSC, TGA, and DMA and the results showed that the incorporation of rGO in electrospun rPET fibers produced rPET/rGO composites. The rPET/rGO composites were then evaluated for possible application as dry electrodes. Moreover, with a preliminary test of numerous volunteers, the rPET/rGO dry electrode showed promising results. The rPET/rGO electrodes showed good performance and applicability to make dry electrodes, and these have applications as dry or wearable electrodes to produce electrochemical sensors.

Correction: Carmo et al. Effects of the Incorporation of Distinct Cations in Titanate Nanotubes on the Catalytic Activity in NOx Conversion. Materials 2021, 14, 2181.

In the original publication, there was a mistake in Figure 1 as published [...].

Amino-Functionalized Titanate Nanotubes: pH and Kinetic Study of a Promising Adsorbent for Acid Dye in Aqueous Solution.

This work reports the functionalization of sodium titanate nanotubes with amine groups obtained from the reaction of titanate nanotubes with [3-(2-Aminoethylamino)propyl]trimethoxysilane, NaTiNT-2NH, and 3-[2-(2-Aminoethylamino)ethylamino]propyltrimethoxysilane, NaTiNT-3NH. It was verified that the crystalline and morphological structures of NaTiNT were preserved after the functionalization, spectroscopies showed that aminosilane interacted covalently with the surface of NaTiNT, and the incorporation of the aminosilane groups on the surface of NaTiNT can be confirmed. The adsorbent matrices NaTiNT-2NH and NaTiNT-3NH were used to remove the anionic dye from remazol blue R (RB) in aqueous medium, and the highest adsorption capacity was around 365.84 mg g-1 (NaTiNT-2NH) and 440.70 mg g-1 (NaTiNT-3NH) in the range of pH 5.0 to 10.0 and the equilibrium time was reached in 210 min (NaTiNT-2NH) and 270 min (NaTiNT-3NH). Furthermore, the Elovich model, which reports the adsorption in heterogeneous sites and with different activation energies in the chemisorption process, was the most appropriate to describe the adsorption kinetics. Thus, these adsorbent matrices can be used as an alternative potential for dye removal RB in aqueous solution.

Nanohydroxyapatite/Titanate Nanotube Composites for Bone Tissue Regeneration.

Strategies for the production of new nanocomposites that promote bone tissue regeneration are important, particularly those that enhance the osteoinduction of hydroxyapatite in situ. Here, we studied and report the synthesis of nanohydroxyapatite and titanate nanotube (nHAp/TiNT) composites formulated at different concentrations (1, 2, 3, and 10 wt % TiNT) by means of a wet aqueous chemical reaction. The addition of TiNT affects the morphology of the nanocomposites, decreasing the average crystallite size from 54 nm (nHAp) to 34 nm (nHAp/TiNT10%), while confirming its interaction with the nanocomposite. The crystallinity index (CI) calculated by Raman spectroscopy and XRD showed that the values decreased according to the increase in TiNT concentration, which confirmed their addition to the structure of the nanocomposite. SEM images showed the presence of TiNTs in the nanocomposite. We further verified the potential cytotoxicity of murine fibroblast cell line L929, revealing that there was no remarkable cell death at any of the concentrations tested. In vivo regenerative activity was performed using oophorectomized animal (rat) models organized into seven groups containing five animals each over two experimental periods (15 and 30 days), with bone regeneration occurring in all groups tested within 30 days; however, the nHAp/TiNT10% group showed statistically greater tissue repair, compared to the untreated control group. Thus, the results of this study demonstrate that the presently formulated nHAp/TiNT nanocomposites are promising for numerous improved bone tissue regeneration applications.

Effects of the Incorporation of Distinct Cations in Titanate Nanotubes on the Catalytic Activity in NOx Conversion.

Effects of the incorporation of Cr, Ni, Co, Ag, Al, Ni and Pt cations in titanate nanotubes (NTs) were examined on the NOx conversion. The structural and morphological characterizations evidenced that the ion-exchange reaction of Cr, Co, Ni and Al ions with the NTs produced catalysts with metals included in the interlayer regions of the trititanate NTs whereas an assembly of Ag and Pt nanoparticles were either on the nanotubes surface or inner diameters through an impregnation process. Understanding the role of the different metal cations intercalated or supported on the nanotubes, the optimal selective catalytic reduction of NOx by CO reaction (SCR) conditions was investigated by carrying out variations in the reaction temperature, SO2 and H2O poisoning and long-term stability runs. Pt nanoparticles on the NTs exhibited superior activity compared to the Cr, Co and Al intercalated in the nanotubes and even to the Ag and Ni counterparts. Resistance against SO2 poisoning was low on NiNT due to the trititanate phase transformation into TiO2 and also to sulfur deposits on Ni sites. However, the interaction between Pt2+ from PtOx and Ti4+ in the NTs favored the adsorption of both NOx and CO enhancing the catalytic performance.

A study of the synthetic methods and properties of graphenes.

Graphenes with varying number of layers can be synthesized by using different strategies. Thus, single-layer graphene is prepared by micromechanical cleavage, reduction of single-layer graphene oxide, chemical vapor deposition and other methods. Few-layer graphenes are synthesized by conversion of nanodiamond, arc discharge of graphite and other methods. In this article, we briefly overview the various synthetic methods and the surface, magnetic and electrical properties of the produced graphenes. Few-layer graphenes exhibit ferromagnetic features along with antiferromagnetic properties, independent of the method of preparation. Aside from the data on electrical conductivity of graphenes and graphene-polymer composites, we also present the field-effect transistor characteristics of graphenes. Only single-layer reduced graphene oxide exhibits ambipolar properties. The interaction of electron donor and acceptor molecules with few-layer graphene samples is examined in detail.

Clinico-Demographic Profile of Patients with Foot Dermatitis: A Cross-Sectional Study with Special Reference to Patch Test Results.

BACKGROUND: Foot dermatitis is a common debilitating dermatological disorder where the eczematous process predominantly involves the feet. AIMS AND OBJECTIVES: To analyze the clinico-demographic profile, type, clinical pattern, and evaluate the role of patch testing to determine the causative factors of foot dermatitis. MATERIALS AND METHODS: Fifty-eight new patients clinically diagnosed with foot dermatitis were subjected to detailed history taking and physical examination. The patients were subjected to patch testing using the Indian standard battery and Indian footwear series (Contact and Occupational Dermatoses Forum of India [CODFI]). RESULTS: Among the 58 patients (mean age 31.48 ± 16.4 years, M:F 1:1.2), the majority (39.7%) presented with allergic contact dermatitis (ACD) followed by irritant contact dermatitis (ICD) (19%), while atopic dermatitis was the least (3.45%). However, 48% of our patients reported a history of atopy. About 43.5% of ACD patients showed a positive patch test reactions to at least one allergen of Indian standard battery and footwear series. Mercaptobenzothiazole (MBT) was the commonest allergen (50%), followed by potassium dichromate (40%), thiuram mix (20%) while paraphenylenediamine was the least common (10%). Dorsum of the foot was affected most commonly (55.17%), followed by toe (46.55%) and sole (41.38%). Scaling was observed in almost 80% of patients followed by crusting. Housewives were affected most commonly followed by students and cement workers. CONCLUSION: Rubber and rubber chemicals were found to be the commonest allergen inciting foot dermatitis. Atopy might be a predisposing factor in these patients. Thus, patch testing is recommended to determine the cause of foot dermatitis and provide suitable treatment.