Integrated transcriptomic and physio-molecular studies unveil the melatonin and PGPR induced protection to photosynthetic attributes in Brassica juncea L. under cadmium toxicity.

Cd is highly mobile, non-essential trace element, that has become serious environmental issue due to its elevated concentration in soil. The present study was taken up to work out salutary effect of melatonin (Mlt) and PGPR ((Pseudomonas putida (Pp), Pseudomonas fluorescens (Pf) in 10 days old Cd stressed (0.3 mM) Brassica juncea L. seedlings. The present work investigated growth characteristics, photosynthetic pigments, secondary metabolites in melatonin-PGPR inoculated B. juncea seedlings. It was backed by molecular studies entailing RT-PCR and transcriptomic analyses. Our results revealed, substantial increase in photosynthetic pigments and secondary metabolites, after treatment with melatonin, P.putida, P. fluorescens in Cd stressed B. juncea seedlings, further validated with transcriptome analysis. Comparative transcriptome analyses identified 455, 5953, 3368, 2238 upregulated and 4921, 430, 137, 27 down regulated DEGs, Cn-vs-Cd, Cd-vs-Mlt, Cd-vs-Mlt-Pp-Pf, Cd-vs-Mlt-Pp-Pf-Cd comparative groups respectively. In depth exploration of genome analyses (Gene ontology, Kyoto encyclopaedia of genes), revealed that Cd modifies the expression patterns of most DEGs mainly associated to photosystem and chlorophyll synthesis. Also, gene expression studies for key photosynthetic genes (psb A, psb B, CHS, PAL, and PSY) suggested enhanced expression in melatonin-rhizobacteria treated Cd stressed B. juncea seedlings. Overall, results provide new insights into probable mechanism of Mlt-PGPR induced protection to photosynthesis in Cd stressed B. juncea plants.

Phytomelatonin maintained chromium toxicity induced oxidative burst in Brassica juncea L. through improving antioxidant system and gene expression.

Chromium (Cr) contamination in soils reduces crop yields and poses a remarkable risk to human and plant system. The main objective of this study was to observe the protective mechanisms of exogenously applied melatonin (Mel- 0.05, 0.1, and 0.15 µM) in seedlings of Brassica juncea L. under Cr (0.2 mM) stress. This was accomplished by analysing the plant's morpho-physiological, biochemical, nuclear, membrane, and cellular characteristics, as well as electrolyte leakage. Superoxide, malondialdehyde, and hydrogen peroxide increased with Cr toxicity. Cr also increased electrolyte leakage. Seedlings under Cr stress had 86.4% more superoxide anion and 27.4% more hydrogen peroxide. Electrolyte leakage increased 35.7% owing to Cr toxicity. B. juncea L. cells with high radical levels had membrane and nuclear damage and decreased viability. Besides this, the activities of the antioxidative enzymes, as POD, APOX, SOD, GST, DHAR, GPOX and GR also elevated in the samples subjected to Cr toxicity. Conversely, the activity of catalase was downregulated due to Cr toxicity. In contrast, Mel reduced oxidative damage and conserved membrane integrity in B. juncea seedlings under Cr stress by suppressing ROS generation. Moreover, the activity of antioxidative enzymes that scavenge reactive oxygen species was substantially upregulated by the exogenous application of Mel. The highest concentration of Mel (Mel c- 0.15 µM) applied showed maximum ameliorative effect on the toxicity caused by Cr. It causes alleviation in the activity of SOD, CAT, POD, GPOX, APOX, DHAR, GST and GR by 51.32%, 114%, 26.44%, 48.91%, 87.51%, 149%, 42.30% and 40.24% respectively. Histochemical investigations showed that Mel increased cell survival and reduced ROS-induced membrane and nuclear damage. The findings showed that Mel treatment upregulated several genes, promoting plant development. Its supplementation decreased RBOH1 gene expression in seedling sunder stress. The results supported the hypothesis that Mel concentrations reduce Cr-induced oxidative burst in B. juncea.

Landfill bacteriology: Role in waste bioprocessing elevated landfill gaseselimination and heat management.

This study delves into the critical role of microbial ecosystems in landfills, which are pivotal for handling municipal solid waste (MSW). Within these landfills, a complex interplay of several microorganisms (aerobic/anaerobic bacteria, archaea or methanotrophs), drives the conversion of complex substrates into simplified compounds and complete mineralization into the water, inorganic salts, and gases, including biofuel methane gas. These landfills have dominant biotic and abiotic environments where various bacterial, archaeal, and fungal groups evolve and interact to decompose substrate by enabling hydrolytic, fermentative, and methanogenic processes. Each landfill consists of diverse bio-geochemical environments with complex microbial populations, ranging from deeply underground anaerobic methanogenic systems to near-surface aerobic systems. These kinds of landfill generate leachates which in turn emerged as a significant risk to the surrounding because generated leachates are rich in toxic organic/inorganic components, heavy metals, minerals, ammonia and xenobiotics. In addition to this, microbial communities in a landfill ecosystem could not be accurately identified using lab microbial-culturing methods alone because most of the landfill's microorganisms cannot grow on a culture medium. Due to these reasons, research on landfills microbiome has flourished which has been characterized by a change from a culture-dependent approach to a more sophisticated use of molecular techniques like Sanger Sequencing and Next-Generation Sequencing (NGS). These sequencing techniques have completely revolutionized the identification and analysis of these diverse microbial communities. This review underscores the significance of microbial functions in waste decomposition, gas management, and heat control in landfills. It further explores how modern sequencing technologies have transformed our approach to studying these complex ecosystems, offering deeper insights into their taxonomic composition and functionality.

Sulfonylpyrazole- and pyrazole-directed ortho-selective C-H functionalization/alkenylation and desulfenylative olefination of aryl(sulfonyl)pyrazoles.

ortho-Selective C-H alkenylation of arenes was achieved using sulfonylpyrazoles and pyrazoles as directing groups, favored by a combination of a Pd(OAc)2 catalyst, Boc-Sar-OH and silver acetate. A wide variety of mono-alkenylated products of aryl-sulfonylpyrazoles and pyrazoles were synthesized with complete site-selectivity under mild reaction conditions. This transformation tolerated several electron-withdrawing and electron-donating groups on the aryl ring and the yields ranged from 52% to 70%, producing highly decorated/valuable alkenylated sulfonylpyrazole and pyrazole derivatives. Amazingly, switching of the oxidant, with the use of AgBF4 in place of AgOAc, offered cinnamic acid derivatives through de-sulfonylation followed by alkenylation at the same position with good yields in the case of aryl-sulfonylpyrazoles. These kinds of molecules have great biological importance and target predictions indicate that they may serve as potential antifungal and anti-tumor agents.

Current studies on the degradation of microplastics in the terrestrial and aquatic ecosystem.

Soil and water are two important basic ecosystems for the survival of different organisms. The excessive microplastic pollutants in soil have been directly discharged into the terrestrial ecosystems. Microplastic pollutants (MPs) constitute a ubiquitous global menace due to their durability, flexibility, and tough nature. MPs posed threat to the sustainability of the ecosystem due to their small size and easy transportation via ecological series resulting in the accumulation of MPs in aquatic and terrestrial ecosystems. After being emitted into the terrestrial ecosystem, the MPs might be aged by oxidative degeneration (photo/thermal), reprecipitation (bioturbation), and hetero-accumulation. The mechanism of adsorption, degradation, and breakdown of MPs into unaffected plastic debris is accomplished by using several biological, physical, and chemical strategies. This review presents the importance of ecosystems, occurrence and sources of MPs, its toxicity, and the alteration in the ecology of the ecosystems. The inhibitory impact of MPs on the ecosystems also documents to unveil the ecological hazards of MPs. Further research is required to study the immobilization and recovery efficiency of MPs on a larger scale.

Pyrazolopyrimidinone Based Selective Inhibitors of PDE5 for the Treatment of Erectile Dysfunction.

Continuing research with our earlier finding of sildenafil based analogs in the search of new inhibitors of PDE5 for erectile dysfunction suggested that there is a scope of modifications at N-methylpiperazine ring with hydrophobic region followed by hydrogen bond donor or acceptor region. However, the leads identified earlier had some limitations like poor pharmacokinetic (PK) profile, low aqueous solubility and poor bioavailability. In this direction, a new series of sildenafil based analogs were designed, synthesized and screened for their PDE5 inhibitory activity. In this series compound 18 was found to have excellent in vitro activity with selectivity towards PDE5 isozyme, also the in vivo activity and pharmacokinetic profile was excellent. The cyp inhibition and CaCO2 permeability was also excellent for compound 18.

Critical review on biogeochemical dynamics of mercury (Hg) and its abatement strategies.

Mercury (Hg) is among the naturally occurring heavy metal with elemental, organic, and inorganic distributions in the environment. Being considered a global pollutant, high pools of Hg-emissions ranging from >6000 to 8000 Mg Hg/year get accumulated by the natural and anthropogenic activities in the atmosphere. These toxicants have high persistence, toxicity, and widespread contamination in the soil, water, and air resources. Hg accumulation inside the plant parts amplifies the traces of toxic elements in the linking food chains, leads to Hg exposure to humans, and acts as a potential genotoxic, neurotoxic and carcinogenic entity. However, excessive Hg levels are equally toxic to the plant system and severely disrupt the physiological and metabolic processes in plants. Thus, a plausible link between Hg-concentration and its biogeochemical behavior is highly imperative to analyze the plant-soil interactions. Therefore, it is requisite to bring these toxic contaminants in between the acceptable limits to safeguard the environment. Plants efficiently incorporate or absorb the bioavailable Hg from the soil thus a constructive understanding of Hg uptake, translocation/sequestration involving specific heavy metal transporters, and detoxification mechanisms are drawn. Whereas recent investigations in biological remediation of Hg provide insights into the potential associations between the plants and microbes. Furthermore, intense research on Hg-induced antioxidants, protein networks, metabolic mechanisms, and signaling pathways is required to understand these bioremediations techniques. This review sheds light on the mercury (Hg) sources, pollution, biogeochemical cycles, its uptake, translocation, and detoxification methods with respect to its molecular approaches in plants.

Phytomicrobiome communications: Novel implications for stress resistance in plants.

The agricultural sector is a foremost contributing factor in supplying food at the global scale. There are plethora of biotic as well as abiotic stressors that act as major constraints for the agricultural sector in terms of global food demand, quality, and security. Stresses affect rhizosphere and their communities, root growth, plant health, and productivity. They also alter numerous plant physiological and metabolic processes. Moreover, they impact transcriptomic and metabolomic changes, causing alteration in root exudates and affecting microbial communities. Since the evolution of hazardous pesticides and fertilizers, productivity has experienced elevation but at the cost of impeding soil fertility thereby causing environmental pollution. Therefore, it is crucial to develop sustainable and safe means for crop production. The emergence of various pieces of evidence depicting the alterations and abundance of microbes under stressed conditions proved to be beneficial and outstanding for maintaining plant legacy and stimulating their survival. Beneficial microbes offer a great potential for plant growth during stresses in an economical manner. Moreover, they promote plant growth with regulating phytohormones, nutrient acquisition, siderophore synthesis, and induce antioxidant system. Besides, acquired or induced systemic resistance also counteracts biotic stresses. The phytomicrobiome exploration is crucial to determine the growth-promoting traits, colonization, and protection of plants from adversities caused by stresses. Further, the intercommunications among rhizosphere through a direct/indirect manner facilitate growth and form complex network. The phytomicrobiome communications are essential for promoting sustainable agriculture where microbes act as ecological engineers for environment. In this review, we have reviewed our building knowledge about the role of microbes in plant defense and stress-mediated alterations within the phytomicrobiomes. We have depicted the defense biome concept that infers the design of phytomicrobiome communities and their fundamental knowledge about plant-microbe interactions for developing plant probiotics.

TBAX/Oxone-Mediated Halogenation of Pyrazoles and Other Heterocycles: An Entry to Important Cross-Coupling Reactions.

A facile, sustainable and eco-friendly protocol has been developed for the halogenation of various heterocycles using TBAX (TBAI/TBAB/TBACl) as halogenating agent, which afforded the products in 90-95% isolated yields. The reaction proceeds with low-cost TBAX and uses greener conditions like EtOH as a solvent and microwave as an alternative energy source for reaction. This protocol has been applied on pyrazoles and extended to different heterocycles like 7-azaindole, indazole, indole and 2-phenylimidazo[1,2-α]pyridines. The gram-scale iodination reaction has also been successfully performed by optimizing conventional heating conditions, which demonstrates its potential applicability in organic synthesis. Further these halogenated pyrazoles have been utilized for different coupling reactions including formation of arylated, alkynylated and sulfenated pyrazoles. However, TBAF mediated fluorination did not work.

Visible-light-promoted iron-catalyzed C-H functionalization of 1,4-naphthoquinones via oxidative coupling with sulfoximines.

A catalytic oxidative addition of sulfoximines to naphthoquinones via C-H functionalization has been achieved using an iron catalytic system, which exhibits good reactivity and high regioselectivity in the presence of visible light. This is the first report offering an efficient protocol for obtaining (naphtho)quinone-sulfoximine hybrid analogs in moderate to good yields with wide scope for both the substrates. This protocol has also been applied on natural products for their modification, including vitamin K3, Juglone and some other modified natural scaffolds as well.

Selective Synthesis of Bis-Heterocycles via Mono- and Di-Selenylation of Pyrazoles and Other Heteroarenes.

The insertion of selenium was achieved in the form of mono-selenides and di-selenides for the preparation of novel bis-heterocyclic compounds. This method is more general and provides scaffold diversity with high yields of products. The concentration-dependent mono- and di-selenylation reaction selectivity was achieved using SeO2 as an efficient selenylating reagent.

Application of melatonin and PGPR alleviates thiamethoxam induced toxicity by regulating the TCA cycle in Brassica juncea L.

Thiamethoxam, a broad spectrum, neonicotinoid insecticide, is used on various crops including Brassica juncea L. to protect from intruding insects such as leaf-hoppers, aphids, thrips and white-flies. Exposure to thiamethoxam causes acute malady such as tumour development, cell apoptosis, liver damage and neurotoxicity. Melatonin is entailed in umpteen developmental processes of plants, including stress responses. The pleiotropic effects of melatonin in modulating plant growth validate it's imperative contribution as multi-regulatory substance. Exiguous information is known about the role of Pseudomonas putida in improving plant growth under thiamethoxam stress. Taking these aspects into consideration the contemporary study investigates the role of melatonin and Pseudomonas putida strain MTCC 3315 in alleviating the thiamethoxam induced toxicity in B. juncea plant. Fourier Transform Infrared Spectroscopy (FTIR) analysis uncloaked that thiamethoxam induced stress primarily affects the protein content of plant as compared to lipids, carbohydrates and cell wall components. Organic acid profiling of the treated samples carried-out by High-Performance Liquid Chromatography (HPLC), reported an upregulation in the level of organic acids, malic acid (110%), citric acid (170%), succinic acid (81%), fumaric acid (40%) and ascorbic acid (55%) in thiamethoxam treated plants compared to the investigational untreated plants. The melatonin treated seedlings grown under thiamethoxam stress, exhibit increased level of malic acid, citric acid, succinic acid, fumaric acid and ascorbic acid by 81%, 0.94%, 11%, 21% and 6% respectively. Further, thiamethoxam stressed plants inoculated with Pseudomonas putida showed stupendous up-regulation by 161% (malic acid), by 14% (citric acid), by 33% (succinic acid), by 30% (fumaric acid), by 100% (oxalic acid) respectively. Lastly, the combinatorial application of melatonin and Pseudomonas putida resulted in prodigious upsurge of malic acid by 165%, succinic acid by 69%, fumaric acid by 42% respectively in contrast to distinct melatonin and Pseudomonas putida treatments. The accumulation of organic acids ascertains the defence against thiamethoxam stress and corresponds to meet the energy generation requirement to skirmish thiamethoxam mediated abiotic stress in Brassica juncea plant.

Activation of lysosomal mediated cell death in the course of autophagy by mTORC1 inhibitor.

Lysosomal biogenesis plays a vital role in cell fate. Under certain conditions, excessive lysosomal biogenesis leads to susceptibility for lysosomal membrane permeabilization resulting in various pathological conditions including cell death. In cancer cells apoptosis machinery becomes dysregulated during the course of treatment, thus allows cancer cells to escape apoptosis. So it is therefore imperative to identify cytotoxic agents that exploit non-apoptotic mechanisms of cell death. Our study showed that pancreatic cancer cells treated with SDS-203 triggered an incomplete autophagic response and a nuclear translocation of transcriptional factor TFEB. This resulted in abundant biosynthesis and accumulation of autophagosomes and lysosomes into the cells leading to their death. It was observed that the silencing of autophagy genes didn't alter the cell fate, whereas siRNA-mediated silencing of TFEB subdued SDS-203 mediated lysosomal biogenesis and associated cell death. Further mouse tumors treated with SDS-203 showed a significant reduction in tumor burden and increased expression of lysosomal markers. Taken together this study demonstrates that SDS-203 treatment triggers non-apoptotic cell death in pancreatic cancer cells through a mechanism of lysosome over accumulation.

Mechanistic investigation of synergistic interaction of tocopherol succinate with a quinoline-based inhibitor of mammalian target of rapamycin.

OBJECTIVES: Cancer monotherapy is associated with various limitations; therefore, combination chemotherapy is widely explored for optimum drug efficacy. In this study, 4-(N-Phenyl-N'-substituted benzenesulfonyl)-6-(4-hydroxyphenyl) quinoline-based mammalian target of rapamycin (mTOR) inhibitor (IIIM-4Q) was investigated in combination with tocopherol succinate (TOS), and the mechanism of cytotoxicity was elucidated. METHODS: The cytotoxic potential of IIIM-4Q and TOS was evaluated in five cell lines. Further, to understand the mechanism of cytotoxicity of IIIM-4Q, TOS and their combination, various studies including morphological analysis using scanning electron microscopy and 6-diamidino-2-phenylindole (DAPI) staining, estimation of reactive oxygen species (ROS) level, measurement of mitochondrial membrane potential (MMP), in-vitro cell migration assay, Western blotting and staining with acridine orange (AO) for autophagy detection were performed. KEY FINDINGS: Investigated combination was synergistic in nature and exhibited greater oxidative stress and mitochondrial dysfunction in pancreatic cancer cells. The migration potential of MIA PaCa-2 cells was significantly mitigated under the influence of this combination, and morphological changes such as chromatin condensation and nuclear blebbing were observed. Also, poly (adenosine diphosphate-ribose) polymerase cleavage and caspase-3 activation were observed in IIIM-4Q and TOS combination-treated cells. CONCLUSIONS: The investigated combination synergistically inhibited proliferation of MIA PaCa-2 cells through simultaneous induction of autophagy followed by apoptosis, and this combination demonstrated potential for further translational studies.

Multiple Facets of Plant-Microbiome Associations in Unlocking the Communication Paradigm through Extracellular Vesicles.

Communication among different species across kingdoms occurs through a chain of regulatory molecules that are transferred around cellular boundaries. These molecules are also crucial for defense, virulence, and pathogenesis. In the past, the transport of proteins in long distance communication was observed, but in the present era, the discovery of extracellular vesicles (EVs) has changed our understanding of molecular communication. EVs are not only involved in cell signaling and immunity but also can transfer information by sRNAs, forming a basis for interactions among a wide variety of organisms. Despite extensive research on EVs in other areas, their role in communication between plants and the plant microbiome has been lacking. EVs are potentially involved in protein trafficking along with the transport of lipids and nucleic acids. Interactions between hosts and their microbiomes may also be mediated by EVs, which can be involved in stress responses, immune surveillance and defense, virulence, and signaling, along with many metabolic activities within plant microbiomes. In this review, we have focused on recent information about the role of EVs and the molecules they transport between hosts and microbes. The connection between biofilms and the generation of EVs is also considered. These findings enhance our knowledge about plant-microbiome interactions in terms of immunity and virulence and challenge the conventional viewpoint of inter-kingdom signaling.

Brassinosteroid Signaling, Crosstalk and, Physiological Functions in Plants Under Heavy Metal Stress.

Brassinosteroids (BRs) are group of plant steroidal hormones that modulate developmental processes and also have pivotal role in stress management. Biosynthesis of BRs takes place through established early C-6 and late C-6 oxidation pathways and the C-22 hydroxylation pathway triggered by activation of the DWF4 gene that acts on multiple intermediates. BRs are recognized at the cell surface by the receptor kinases, BRI1 and BAK1, which relay signals to the nucleus through a phosphorylation cascade involving phosphorylation of BSU1 protein and proteasomal degradation of BIN2 proteins. Inactivation of BIN2 allows BES1/BZR1 to enter the nucleus and regulate the expression of target genes. In the whole cascade of signal recognition, transduction and regulation of target genes, BRs crosstalk with other phytohormones that play significant roles. In the current era, plants are continuously exposed to abiotic stresses and heavy metal stress is one of the major stresses. The present study reveals the mechanism of these events from biosynthesis, transport and crosstalk through receptor kinases and transcriptional networks under heavy metal stress.

Oxone-DMSO Triggered Methylene Insertion and C(sp2)-C(sp3)-H-C(sp2) Bond Formation to Access Functional Bis-Heterocycles.

Metal-free insertion of a methylene group was achieved for the construction of a new C(sp2)-C(sp3)-H-C(sp2) bond in order to prepare novel bis-heterocyclic scaffolds. The complete mechanistic investigations included experimental study and DFT calculations, and various symmetric and unsymmetric bis-pyrazoles as well as other pyrazole-based bis-heterocyclic molecules were prepared in moderate to high yields. Further modification of the bridged methylene group in the unsymmetric pyrazoles generated a chiral center to extend the scope of this method.

Direct N-heterocyclization of hydrazines to access styrylated pyrazoles: synthesis of 1,3,5-trisubstituted pyrazoles and dihydropyrazoles.

A microwave-assisted method has been developed for the synthesis of tri-substituted pyrazoles via direct N-heterocyclization of hydrazines with metal-acetylacetonate and -dibenzylideneacetonate without using any base or additives. Most importantly, the synthesis of 1-aryl-5-phenyl-3-styryl-1H-pyrazoles was achieved in a single step using hydrochloride salt of various phenylhydrazines and this is the first report for direct construction of these molecules. The reaction medium and microwave conditions play a critical role for their selective product formation during the reaction. The present reaction explored the usage of metal-diketonic complexes as reaction substrates providing acetylacetone and dibenzylideneacetone moieties to directly participate in cyclization with hydrazines to form the corresponding pyrazoles in excellent yields. The present protocol introduces the important N-heterocyclic moieties in the final structures, giving the reaction great applications from a medicinal chemistry perspective, particularly in the late stage modification strategies in drug discovery.

Antioxidant activity of essential oil and extracts of Valeriana jatamansi roots.

Valeriana jatamansi is an indigenous medicinal plant used in the treatment of a number of diseases. In the present study, chemical composition of the essential oil was determined by GC-MS. Seven major components were identified in Valeriana jatamansi essential oil, namely, ß-vatirenene, ß-patchoulene, dehydroaromadendrene, ß-gurjunene, patchoulic alcohol, ß-guaiene, and α-muurolene. Methanolic, aqueous, and chloroform extracts of Valeriana jatamansi roots were also prepared and analyzed for their polyphenols and flavonoid content. Antioxidant activity of essential oil and different extracts of Valeriana jatamansi roots was determined by DPPH radical scavenging and chelation power assay. A linear correlation has been obtained by comparing the antioxidant activity and polyphenols and flavonoid content of the extracts. Results indicated that antioxidant activity of methanolic extract could be attributed to the presence of rich amount of polyphenols and flavonoid. Essential oil of Valeriana jatamansi roots showed moderate antioxidant activity.