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Entomopathogenic nematodes (EPNs) of the genera Heterorhabditis and Steinernema kill insects with the help of their symbiotic bacteria. They are widely used as biocontrol agents to manage insect pests of crops. The infective juveniles (IJ) of EPNs are isolated from soil by insect baiting technique, which is labour-intensive, time-consuming, wasteful, and inefficient. Here, we present loop-mediated isothermal amplification (LAMP) assays for rapid detection of Heterorhabditis spp. (Het-LAMP) and Steinernema spp. (Ste-LAMP) from total soil DNA. The primers for Het-LAMP and Ste-LAMP were designed using ITS and 18S rDNA regions of genomic DNA. The LAMP reactions could be completed in 60 min, at 66 °C and 68 °C, respectively, followed by termination at 85 °C for 5 min. The assays were highly sensitive and could detect up to 0.02 picograms of Heterorhabditis DNA and 96 picograms of Steinernema DNA in a 25 μl reaction. Both the assays were specific for the target nematode species and detected the presence of a single IJ in the total DNA extracted from 250 mg of soil. The assays developed in this study would be of immense utility for the efficient detection and identification of native EPNs in large-scale surveys. These assays are amenable to automation and could be used to develop convenient detection kits for point-of-service diagnosis of EPNs in the field without the need for a trained and experienced personnel.

Plants catalyze the biosynthesis of a large number of non-protein amino acids, which are usually toxic for other organisms. In this review, the chemistry and metabolism of N-heterocyclic non-protein amino acids from plants are described. These N-heterocyclic non-protein amino acids are composed of β-substituted alanines and include mimosine, β-pyrazol-1-yl-L-alanine, willardiine, isowillardiine, and lathyrine. These β-substituted alanines consisted of an N-heterocyclic moiety and an alanyl side chain. This review explains how these individual moieties are derived from their precursors and how they are used as the substrate for biosynthesizing the respective N-heterocyclic non-protein amino acids. In addition, known catabolism and possible role of these non-protein amino acids in the actual host is explained.

The Gram-negative bacteria Photorhabdus lives in a symbiotic relationship with the insect-pathogenic Heterorhabditis nematodes and produces numerous hydrolytic enzymes, secondary metabolites and protein toxins. Seven Photorhabdus strains were previously isolated from the Heterorhabditis nematodes collected from different geographical regions of India. The strains IARI-SGMG3, IARI-SGHR2, IARI-SGHR4, IARI-SGMS1 and IARI-SGGJ2 were identified as P. akhurstii, whereas IARI-SGLDK1 and IARI-SGHP1 were identified as P. laumondii subsp. laumondii and P. laumondii subsp. clarkeii, respectively. A new and previously unreported 35 kDa molecular weight protein toxin 'Galtox' was identified from these Photorhabdus strains. The nucleotide sequences of the toxin gene from seven Photorhabdus strains were PCR amplified, sequenced, cloned into pET protein expression vector, and the protein toxin was expressed and purified. The Galtox sequence from various strains showed variations in sequence and toxicity against Galleria mellonella. The injection of purified Galtox protein into the 4th instar larvae showed median lethal dose (LD50) values of 2.39–26.08 ng toxin/g G. mellonella bodyweight after 48 h. The protein injection killed the insects quickly and exhibited a median lethal time (LT50) of 12–60 h when injected at the rate of 3.1–31.2 ng toxin/g G. mellonella bodyweight. Galtox protein sequence analysis indicated similarity to several bacterial toxin-related protein domains, such as 6rgnA domain of Bordetella membrane targeting toxin BteA, 6gy6 domain of Xenorhabdus α-Xenorhabdolysins, 4mu6A and 4xa9a domains similar to effector protein LegC3 from Legionella pneumophila and 1cv8.1 domain of staphylococcal cysteine proteinase staphopain B. The mode of action of Galtox needs to be understood to enable its use for the management of agricultural insect-pests.

The experiment on 108 treatments of wheat along with 80 F1’s, 24 parental lines (20 females + 4 males) and four standard variety (KH-65, KRL-1702, KRL-1714, NW-1014) of wheat was conducted to work out the genetic variability, heritability and genetic advance effects of their various attributes on grain yield. The high estimates (>15%) of phenotypic (PCV) and genotypic (GCV) coefficients of variation were recorded in case of tiller per plant (PCV=16.37%, GCV=9.14%). High estimates of broad sense heritability (> 75%) were recorded for plant height (80.00%) and Tiller/plant, in sodic soil showed moderate estimate of genetic advance in percent of mean (10-20%). High heritability and genetic advance indicate that the additive nature of gene action and reliability of those characters for selection and emerged as ideal traits for improvement through selection.

RNA exosome is a highly conserved ribonuclease complex essential for RNA processing and degradation. Bi-allelic variants in exosome subunits EXOSC3, EXOSC8 and EXOSC9 have been reported to cause pontocerebellar hypoplasia type 1B, type 1C and type 1D, respectively, while those in EXOSC2 cause short stature, hearing loss, retinitis pigmentosa and distinctive facies. We ascertained an 8-months-old male with developmental delay, microcephaly, subtle dysmorphism and hypotonia. Pontocerebellar hypoplasia and delayed myelination were noted on neuroimaging. A similarly affected elder sibling succumbed at the age of 4-years 6-months. Chromosomal microarray returned normal results. Exome sequencing revealed a homozygous missense variant, c.104C > T p.(Ser35Leu) in EXOSC1 (NM_016046.5) as the possible candidate. In silico mutagenesis revealed loss of a polar contact with neighboring Leu37 residue. Quantitative real-time PCR indicated no appreciable differences in EXOSC1 transcript levels. Immunoblotting and blue native PAGE revealed reduction in the EXOSC1 protein levels and EXO9 complex in the proband, respectively. We herein report an individual with the bi-allelic variant c.104C>T p.(Ser35Leu) in EXOSC1 and clinical features of pontocerebellar hypoplasia type 1. Immunoblotting and blue native PAGE provide evidence for the pathogenicity of the variant. Thus, we propose EXOSC1 as a novel candidate gene for pontocerebellar hypoplasia.

Coronavirus disease 2019 (COVID-19) caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has caused a global pandemic. RNA-dependent RNA polymerase (RdRp) is the key component of the replication or transcription machinery of coronavirus. Therefore SARS-CoV-2-RdRp has been chosen as an important target for the development of antiviral drug(s). During the early pandemic of the COVID-19, chloroquine and hydroxychloroquine were suggested by the researchers for the prevention or treatment of SARS-CoV-2. In our study, the antimalarial compounds have been screened and docked against SARS-CoV-2-RdRp (PDB ID: 7BTF), and it was observed that the antimalarials chloroquine, hydroxychloroquine, and amodiaquine exhibit good affinity. Since the crystal structure of SARS-CoV-2-RdRp with its substrate is not available, poliovirus-RdRp crystal structure co-crystallized with its substrate ATP (PDB ID: 2ILY) was used as a reference structure. The superimposition of SARS-CoV-2-RdRp and poliovirus-RdRp structures showed that the active sites of both of the RdRps superimposed very well. The amino acid residues involved in the binding of ATP in the case of poliovirus-RdRp and residues involved in binding with the antimalarial compounds with SARS-CoV-2-RdRp were compared. In both cases, the conserved residues were found to be involved in establishing the interactions. The MMGBSA and molecular dynamic simulation studies were performed to strengthen our docking results. Further residues involved in binding of antimalarials with SARS-CoV-2-RdRp were compared with the residues involved in the SARS-CoV-2-RdRp complexed with remdesivir [PDB ID: 7BV2]. It was observed that co-crystallized remdesivir and docked antimalarials bind in the same pocket of SARS-CoV-2 -RdRp. Communicated by Ramaswamy H. Sarma

Cultivated fenugreek Trigonella corniculata L, also known as kasuri methi or nagauri paan is a popular spice crop and widely grown in the state of Rajasthan, India. Galls and root-knot nematodes were observed on the roots of T. corniculata collected from Naguar district and its surrounding areas in the state of Rajasthan. The nematode DNA isolated from the sample was subjected to PCR by using species-specific primers for the common root-knot nematode species. Molecular identification confirmed the infestation of Meloidogyne enterolobii on T. corniculata. This is the first report of M. enterolobii infestation on T. corniculata from India.

Background Txp40, a 37 kDa protein, previously characterized from the Gram-negative bacterium Photorhabdus akhurstii (symbiotically associates with insect-parasitic nematode, Heterorhabditis indica), conferred insecticidal activity against Galleria mellonella. Here, the biological activity of Txp40 was evaluated against economically important insects, including Helicoverpa armigera, Spodoptera litura and S. exigua. Results When both intra-hemocoel injected and orally fed to test insects, comparatively greater oral LD50 (187.7-522 ng g-1 ) than injection LD50 (32.33-150.6 ng g-1 ) was obtained with Txp40 derived from P. akhurstii strain IARI-SGMG3. Injection of purified Txp40 caused a dose-dependent reduction in the total circulatory hemocytes and hemocyte viability of fourth-instar larvae of the test insects at 12 h post incubation; unlike healthy cells toxin-treated ones displayed aggregated distribution. Injection of Txp40 significantly elevated the phenoloxidase activity of insect hemolymph, which potentially led to unrestrained melanization reaction and ultimately larval death. Histological analyses showed the primary site of action of Txp40 in the insect midgut. Extensive damage to midgut epithelium 24 h after injection of the Txp40 explains the access of the toxin from hemocoel to midgut via leaky septate junctions. In silico analyses suggested that Txp40 can potentially interact with H. armigera midgut receptor proteins cadherin, ATP-binding cassettes, aminopeptidase N1 and alkaline phosphatase to exert toxicity. Conclusion We propose Txp40 as an attractive alternative to Cry toxins of Bacillus thuringiensis, the transgenic expression of which is reported to cause resistance development in insects. © 2019 Society of Chemical Industry.

Root-knot nematodes (Meloidogyne spp.) infect a large number of crops including guava. We investigated a population of Meloidogyne sp. infecting guava in the Coimbatore region of Tamil Nadu, India for identification and species confirmation. Detailed morphological and morphometric observations based on second-stage juveniles, males, females, and perineal patterns showed resemblance of the isolated population with the original and subsequent descriptions of M. enterolobii. Isozyme analysis of the young egg-laying females displayed the characteristic esterase phenotype pattern similar to that of M. enterolobii. Additionally, the identity of the nematode population was further validated by M. enterolobii specific SCAR marker and ITS rDNA. Recently published reports on the occurrence and morphological descriptions of M. enterolobii from India are largely incongruent with the original and subsequent redescriptions of the species. Here, we present the most comprehensive morphology and morphometrics of an Indian population of M. enterolobii for its authentic identification.

High-throughput genotyping has become more convenient and cost-effective due to recent advancements in next-generation sequencing (NGS) techniques. Numerous approaches exploring sequencing advances for genotyping have been developed over the past decade, which includes different variants of genotyping-by-sequencing (GBS), and restriction-site associated DNA sequencing (RAD-seq). Most of these methods are based on the reduced representation of the genome, which ultimately reduces the cost of sequencing by many folds. However, continuously lowering the cost of sequencing makes it more convenient to use whole genome-based approaches. In this regard, skim sequencing, where low coverage whole-genome sequencing is used for the identification of large numbers of polymorphic markers cost-effectively. In the present review, we have discussed recent technological advancements, applicability, and challenges of skim sequencing-based genotypic approaches for crop improvement programmes. Skim sequencing is being extensively used for genotyping in diverse plant species and has a wide range of applications, particularly in quantitative trait loci (QTL) mapping, genomewide association studies (GWAS), fine genetic map construction, and identification of recombination and gene conversion events in various breeding programmes. The cost-effectiveness, simplicity, and genomewide coverage will increase the application of skims sequencing-based genotyping. The article summarizes the protocol, uses, bioinformatics tools, its application, and future prospects of skim sequencing in crop improvement.

This chapter focuses on target traits for drought stress, progress in mapping for drought tolerance-associated genes/QTLs identification and expression studies and introgression strategies followed by the possibilities of integrating the concept of speed breeding in maize drought breeding programmes for better utilization of wild relatives.

Cyst nematodes of the species Globodera rostochiensis and G. pallida are devastating parasites of the potato crop. Early detection of cyst nematodes in the field is critical for adopting an appropriate management strategy. A specific and sensitive loop-mediated isothermal amplification (LAMP) assay using four oligonucleotide primers has been developed to amplify the internal transcribed spacer region (ITS) of ribosomal DNA of potato cyst nematode G. rostochiensis. The PCN-LAMP reaction could be completed within 75 min at 68 °C followed by termination at 85 °C for 7 min. The primers exhibited specificity for G. rostochiensis and did not detect any other tested genera of plant parasitic or entomopathogenic nematodes. LAMP reaction was highly sensitive, suitable for crude genomic DNA and could successfully detect G. rostochiensis DNA up to femtogram quantity. This assay is rapid, cost effective and requires minimal instrumentation. It will facilitate the detection of G. rostochiensis at field and point-of-care labs and help in the interception of infested plant material/soil samples at quarantine stations independent of a professional nematologist.

Biallelic loss of function variants in TRIP11 encoding for the Golgi microtubule-associated protein 210 (GMAP-210) cause the lethal chondrodysplasia achondrogenesis type 1A (ACG1A). Loss of TRIP11 activity has been shown to impair Golgi structure, vesicular transport and results in loss of IFT20 anchorage to the Golgi that is vital for ciliary trafficking and ciliogenesis. Here we report four fetuses, two each from two families, who were ascertained antenatally with ACG1A. Affected fetuses in both families are homozygous for the deep intronic TRIP11 variant, c.5457+81T>A that was found in a shared region of homozygosity. This variant was found to cause aberrant transcript splicing and the retention of 77 base-pairs of intron 18. The TRIP11 messenger RNA and protein levels were drastically reduced in fibroblast cells derived from one of the affected fetuses. Using immunofluorescence we also detected highly compacted Golgi apparatus in affected fibroblasts. Further we observed significant reduction in the frequency of ciliated cells and in the length of primary cilia in subject derived cell-lines, not reported so far in patient cells with TRIP11 null or hypomorphic variants. Our findings illustrate how pathogenic variants in intronic regions of TRIP11 can impact transcript splicing, expression and activity resulting in ACG1A. This article is protected by copyright. All rights reserved.

Amyna axis (Lepidoptera: Erebidae), Chrysodeixis spp. ( C. acuta and C. eriosoma ) and Spodoptera spp. ( S. exigua and S. litura ) (Lepidoptera: Noctuidae) are important polyphagous insect-pests of leguminous crops. A mermithid (Nematoda: Mermithidae) nematode was found parasitizing the larval stages of these insects in the western Indian state of Rajasthan. Morphological and molecular analysis suggests that the nematode might be a species of the genus Hexamermis . The average length of post-parasitic nematode juveniles was 15 cm, and the average greatest body width was 1.5 mm. The vulva was median (V% = 48) without vulval flap. Approximately 100-125 μm long caudal appendages were present on tail. The cross-section revealed the presence of six hypodermal chords at the mid body region, and stichosome was present. The 18S rDNA sequence showed 99% similarity to mermithid nematode. A phylogenetic analysis of 18S rDNA sequences of mermithids resulted in identification of a new clade “Lineage 3” which included representatives of Mermis sp., Isomermis sp., Limnomermis sp., and Pheromermis sp. This is the first report of natural mermithid parasitism of A. axis and Chrysodeixis spp. along with its molecular characterization.

Peptidyl arginine deiminase-4 (PAD4) is indispensable for generation of neutrophil extracellular traps (NETs), which can provide antimicrobial effects during host innate immune response; however, the role of PAD4 against gastrointestinal infection is largely unknown. Herein, we challenged PAD4-deficient (Pad4-/-) mice and wild-type (WT) littermates with Citrobacter rodentium (CR), and investigated bacteria clearance and gut pathology. Luminal colonization of CR in Pad4-/- mice peaked between 11-14 days post-infection, whereas WT mice suppressed the infection by 14 days. We demonstrated that Pad4-/- mice were unable to form NETs, whereas WT mice showed increased NETs formation in the colon during infection. Pad4-/- mice showed aggravated CR-associated inflammation as indicated by elevated systemic and colonic pro-inflammatory markers. Histological analysis revealed that transmissible colonic hyperplasia, goblet cell depletion, and apoptotic cell death were more pronounced in the colon of CR-infected Pad4-/- mice. Treating WT mice with deoxyribonuclease I, which can disrupt NETs generation, recapitulated the exacerbated CR infection and gut pathology associated with the loss of PAD4. Administration of the PAD4 inhibitor, Cl-amidine also aggravated CR infection, but to a lesser extent. Taken together, our findings highlight the importance of PAD4 in the mucosal clearance of CR and in resolving gut-associated inflammation.

Summary A new species of pin nematode, Paratylenchus jasmineae sp. n., was isolated and described from the rhizosphere of jasmine in India. The new species can be diagnosed by having a relatively larger body length of 248-355 μm, stylet length of 24.6-28.5 μm, lateral field with four lateral lines, the presence of small post-vulval uterine sac and absence of advulval flaps in the females. Additional morphological characters included continuous lip region with small projected submedian lobes, large and oblong-shaped spermatheca, the presence of prevulval swelling, and arcuate conoid tail with finely rounded terminus. Molecular characterisation of the species was carried out based on internal transcribed spacer sequence (ITS rRNA) and D2-D3 expansion segment of 28S rRNA subunit. Phylogenetic analyses based on ITS and D2-D3 molecular markers resulted in clear separation of P. jasmineae sp. n. from other Paratylenchus species.

Summary A population of a Hemicriconemoides species was found from the rhizosphere of sugarcane in the Meerut district, Uttar Pradesh, India. Preliminary morphological studies led to the identification of the species as H. rosae. Further morphological and morphometric characterisation of the species was carried out by light microscopy. The gross morphology was consistent with the description of H. rosae except for some deviations in measurements. Here we present an amended description of the species together with illustrations and additional morphometric and morphological characters. The diagnostic relationship of H. rosae is given based on morphological and molecular information and a comparative study of paratype and holotype females. In addition, ITS rRNA and D2-D3 sequences were used for the construction of evolutionary relationship by Bayesian inference. The present study provides molecular data and a redescription of H. rosae collected from the rhizosphere of sugarcane in India.

The global increasing burden of non-communicable diseases such as cancer and diabetes has generated an immediate need for the development of sustainable drugs. Transition metal complexes with their versatility can offer excellent drug candidature potency. With very few reports on the non-toxic and safe drug candidatures of transition metal complexes, we have hereby attempted to explore the bioactivity of newly synthesized Fe3+ and Co3+ complexes tethered with an ONS donor ligand core. The new compounds, [FeIII(OH2)(OMe)(Hhpdbal-sbdt)2] (1) and [CoIII(OH2)(Cl)(Hhpdbal-sbdt)2] (2) were characterized using the usual analytical and spectroscopic techniques. The human cervical cancer cell line (HeLa) was used as the model cancer cell line against which the anticancer activities of compounds were evaluated. Both compounds 1 and 2 exhibited significant antineoplastic behavior with IC50 values of 4.61 μM and 4.70 μM, respectively. Human liver carcinoma cells (HePG2) were used as model diabetic or insulin resistant cells to study the glucose uptake activity of the compounds. The cobalt complex 2 demonstrated excellent glucose uptake (89.83%). To rationalize and compliment the biological significance of these compounds, the cytotoxicity or the safety profile was addressed against the healthy human kidney cell line (HEK-293), and from this study, the CC50 values of complexes 1 and 2 were calculated to be 412.68 μM and 473.17 μM, respectively, which reflected the non-toxic nature of the candidates. Furthermore, the BSA protein interactions of 1 and 2 were preliminarily interpreted by studying binding thermodynamics, quenching and binding interactions, which suggested facile binding between compounds and BSA.

Background Southern root-knot nematode Meloidogyne incognita (Kofoid and White, 1919), Chitwood, 1949 is a key pest of agricultural crops. Pasteuria penetrans is a hyperparasitic bacterium capable of suppressing the nematode reproduction, and represents a typical coevolved pathogen-hyperparasite system. Attachment of Pasteuria endospores to the cuticle of second-stage nematode juveniles is the first and pivotal step in the bacterial infection. RNA-Seq was used to understand the early transcriptional response of the root-knot nematode at 8 h post Pasteuria endospore attachment. Results A total of 52,485 transcripts were assembled from the high quality (HQ) reads, out of which 582 transcripts were found differentially expressed in the Pasteuria endospore encumbered J2 s, of which 229 were up-regulated and 353 were down-regulated. Pasteuria infection caused a suppression of the protein synthesis machinery of the nematode. Several of the differentially expressed transcripts were putatively involved in nematode innate immunity, signaling, stress responses, endospore attachment process and post-attachment behavioral modification of the juveniles. The expression profiles of fifteen selected transcripts were validated to be true by the qRT PCR. RNAi based silencing of transcripts coding for fructose bisphosphate aldolase and glucosyl transferase caused a reduction in endospore attachment as compared to the controls, whereas, silencing of aspartic protease and ubiquitin coding transcripts resulted in higher incidence of endospore attachment on the nematode cuticle. Conclusions Here we provide evidence of an early transcriptional response by the nematode upon infection by Pasteuria prior to root invasion. We found that adhesion of Pasteuria endospores to the cuticle induced a down-regulated protein response in the nematode. In addition, we show that fructose bisphosphate aldolase, glucosyl transferase, aspartic protease and ubiquitin coding transcripts are involved in modulating the endospore attachment on the nematode cuticle. Our results add new and significant information to the existing knowledge on early molecular interaction between M. incognita and P. penetrans. Electronic supplementary material The online version of this article (10.1186/s12864-018-5230-8) contains supplementary material, which is available to authorized users.

The rice root-knot nematode Meloidogyne graminicola is a major biotic stress for the rice crop under upland, rain-fed lowland and irrigated cultivation conditions. Here, we present an improved draft genome assembly of M. graminicola IARI strain using the long-read sequencing approach (PacBio Sequel platform). The assembled genome size was 36.86 Mb with 514 contigs and N50 value of 105 kb. BUSCO estimated the genome to be 88.6% complete. Meloidogyne graminicola genome contained 17.83% repeat elements and showed 14,062 protein-coding gene models, 4,974 conserved orthologous genes, 561 putative secreted proteins, 49 RNAi pathway genes, 1,853 proteins involved in pathogen-host interactions, 1,575 carbohydrate-active enzymes, and 32,138 microsatellites. Five of the carbohydrate-active enzymes were found only in M. graminicola genome and were not present in any other analysed root-knot nematode genome. Together with the previous two genome assemblies, this improved genome assembly would facilitate comparative and functional genomics for M. graminicola.

Resistance to rice root-knot nematode Meloidogyne graminicola in a mutant rice line is suggested to be conferred by higher expression of several genes putatively involved in damage-associated molecular pattern recognition, secondary metabolite biosynthesis including phytoalexins, and defence-related genes. Meloidogyne graminicola has emerged as the most destructive plant–parasitic nematode disease of rice (Oryza sativa L.). Genetic resistance to M. graminicola is one of the most effective methods for its management. A M. graminicola-resistant O. sativa ssp. indica mutant line-9 was previously identified through a forward genetic screen (Hatzade et al. Biologia 74:1197–1217, 2019). In the present study, we used RNA-Sequencing to investigate the molecular mechanisms conferring nematode resistance to the mutant line-9 compared to the susceptible parent JBT 36/14 at 24 h post-infection. A total of 674 transcripts were differentially expressed in line-9. Early regulation of genes putatively related to nematode damage-associated molecular pattern recognition (e.g., wall-associated receptor kinases), signalling [Nucleotide-binding, Leucine-Rich Repeat (NLRs)], pathogenesis-related (PR) genes (PR1, PR10a), defence-related genes (NB-ARC domain-containing genes), as well as a large number of genes involved in secondary metabolites including diterpenoid biosynthesis (CPS2, OsKSL4, OsKSL10, Oscyp71Z2, oryzalexin synthase, and momilactone A synthase) was observed in M. graminicola-resistant mutant line-9. It may be suggested that after the nematode juveniles penetrate the roots of line-9, early recognition of invading nematodes triggers plant immune responses mediated by phytoalexins, and other defence proteins such as PR proteins inhibit nematode growth and reproduction. Our study provides the first transcriptomic comparison of nematode-resistant and susceptible rice plants in the same genetic background and adds to the understanding of mechanisms underlying plant-nematode resistance in rice.

The ecosystem functioning of any functional niche is largely dependent on the structure and organization of the inhabiting microbial communities. The microbial communities often display dynamic organization which is under constant exposure to diverse abiotic stress conditions. Such stress conditions play a major role in shaping and influencing the microbial community organization and its diversity. Microorganisms, which represent the earliest life forms, have undergone the longest evolutionary period resulting in the acquisition and development of capabilities to withstand extreme stress conditions. Besides their resilience nature against various abiotic stress conditions, microbes also display adaptability by rapid mutation to counter the stress conditions. As different microbes have different capabilities to tolerate any stress condition, the stress condition often favors the enrichment of microbes which display tolerance to the exposed condition. In this chapter, we summarized various abiotic stress conditions, including temperature, salt, drought, waterlogging, and metal toxicity stress, and how they influence the structure and diversity of the inhabiting microbial community structure and diversity. Various mechanisms employed by microorganisms to withstand these abiotic stress conditions are also described in the chapter.

Metabolic disorders have been increasing at an alarming rate, and one such example of metabolic disorder is type 2 diabetes mellitus (T2DM). Unregulated gluconeogenesis in T2DM results in increased hepatic glucose output that causes fasting and postprandial hyperglycaemia. Extensive proofs have shown that the downregulation of the key rate-limiting enzyme phosphoenolpyruvate carboxykinase-1 (PCK-1) of gluconeogenesis improved glucose homeostasis in vivo. In the present study, we have synthesized and characterized liver-specific stearic acid conjugated octaarginine (StA-R8) functionalized 4arm-2K-PEGamineylated graphene oxide nanosheets (GPR8) for the delivery of siRNA against PCK-1 in T2DM C57BL/6 mice. We found that a single intravenous administration of siRNA (3 mg/kg BW) conjugated to GPR8 (GPR8:PCK-1siRNA(3 mg/kg BW) conjugate) in an optimized N/P ratio exploited as a therapeutic nanoformulation maintained glucose homeostasis for nearly 4 weeks in the T2DM mice. Efficient silencing of PCK-1 in T2DM liver tissue increased the phosphorylation of serine-256 of FOXO-1, thus showing a marked decrease in hepatic gluconeogenesis. Gluconeogenesis control and consequently glucose output from the liver furthermore partially enhanced liver and muscle insulin sensitivity results in the stimulation of the insulin/AKT-2 signaling pathway which indirectly restored glucose homeostasis in the treated T2DM group. Our therapeutic nanoformulation also improved glycogen storage in the liver and membrane translocation of GLUT4 in the muscle of the treated T2DM group. In conclusion, GPR8:PCK-1siRNA (3 mg/Kg BW) restored glucose homeostasis by controlling the hepatic glucose production and improved peripheral insulin sensitivity as a consequence of reduced hyperglycemia. Thus, the current approach offered an alternative strategy for the therapeutics for T2DM.

The impact of increasing human population, rising food demand, and adverse effects of climate change, viz., changing rainfall pattern, rising temperature, biotic-abiotic stresses, etc., has tremendously affected global food security. In addition, increased anthropogenic inputs from urbanization, industrialization, as well as outrageous use of chemical fertilizers and pesticides have posed a severe threat to the sustainability of the agroecosystems. For many decades, the use of chemical pesticides against insect and microbial pests has become an integrative part of agriculture and contributed significantly to the crop improvement. But, their long-term persistence, cytotoxicity, and microbial resistance have resulted negative impact on the biosphere, thus creating pollution of diverse ecosystems, land degradation, and biodiversity losses. For the last two decades, alternate pest management strategies have become the new avenues for controlling pest and diseases in a greener, safer, and eco-friendly manner. The use of biological control agents (termed as biocides) such as both microbe- and plant-based formulations has been known to be the major emerging tool in crop disease/pest management and appealing alternative to the chemical pesticide in sustainable agriculture. Biopesticides employ the use of naturally occurring substances, i.e., living organisms (natural enemies) or their products (phytochemicals, microbial products) or by-products (semiochemicals) that control pests by nontoxic mechanisms, with high targeted activity against causal agents (insects, fungi, weeds, viruses, nematodes, etc.), and nonpersistence in the environment. The use of biopesticide alone or in combination with agrochemicals has become the new tool in crop protection as a part of biointensive integrated pest management (IPM) strategies. Although biopesticides are slowly substituting the chemical pesticides with great promise, its use to the desired extent is lacking; hence insight on such biological agents is a prerequisite. In this chapter, we have summarized the sources of biopesticides, their plant protective mechanisms (mode of action), availability, and status in India, as well as some critical pros and cons of its use.

Precise monitoring of the enzyme activity by a suitable modulator is one of the very fundamental aspects of drug designing that provides the opportunity to overcome the challenges of several diseases. Herein, inhibition of human Topoisomerase IIα enzyme which serves as a potential target site for several anti-cancer drugs is demonstrated by using ultra-small size gold nanoclusters (Au NCs) with the dimension comparable with size of the active site of the enzyme. Molecular dynamics simulation results demonstrate that the Au NCs strongly interact with the human Topo IIα enzyme at its active site or allosteric site depending on forms of enzyme. Additionally, binding energy and interaction profile provides the molecular basis of understanding of interactions of ultra-small size Au NCs and human Topo IIα enzyme. Enthalpy change (ΔH) and binding constant (K) are measured based on a sequential binding model of the Au NCs with the enzyme as demonstrated by the ITC study. Moreover, the in-vitro inhibition study of the catalytic activity of the enzyme and gel electrophoresis indicates that the ultra-small size Au NCs may be used as a potent inhibitor of human Topo IIα enzyme.

Primary cilia are non-motile sensory antennae present on most vertebrate cell surfaces. They serve to transduce and integrate diverse external stimuli into functional cellular responses vital for development, differentiation and homeostasis. Ciliary characteristics, such as length, structure and frequency are often tailored to distinct differentiated cell states. Primary cilia are present on a variety of skeletal cell-types and facilitate the assimilation of sensory cues to direct skeletal development and repair. However, there is limited knowledge of ciliary variation in response to the activation of distinct differentiation cascades in different skeletal cell-types. C3H10T1/2, MC3T3-E1 and ATDC5 cells are mesenchymal stem cells, preosteoblast and prechondrocyte cell-lines, respectively. They are commonly employed in numerous in vitro studies, investigating the molecular mechanisms underlying osteoblast and chondrocyte differentiation, skeletal disease and repair. Here we sought to evaluate the primary cilia length and frequencies during osteogenic differentiation in C3H10T1/2 and MC3T3-E1 and chondrogenic differentiation in ATDC5 cells, over a period of 21 days. Our data inform on the presence of stable cilia to orchestrate signaling and dynamic alterations in their features during extended periods of differentiation. Taken together with existing literature these findings reflect the occurrence of not only lineage but cell-type specific variation in ciliary attributes during differentiation. These results extend our current knowledge, shining light on the variabilities in primary cilia features correlated with distinct differentiated cell phenotypes. It may have broader implications in studies using these cell-lines to explore cilia dependent cellular processes and treatment modalities for skeletal disorders centered on cilia modulation.

Plant–parasitic root-knot nematode Meloidogyne incognita uses an array of effector proteins to establish successful plant infections. Mi-msp-1 and Mi-msp-20 are two known effectors secreted from nematode subventral oesophageal glands; Mi-msp-1 being a putative secretory venom allergen AG5-like protein, whereas Mi-msp-20 is a pioneer gene with a coiled-coil motif. Expression of specific effector is known to cause disturbances in the expression of other effectors. Here, we used RNA-Seq to investigate the pleiotropic effects of silencing Mi-msp-1 and Mi-msp-20. A total of 25.1–51.9 million HQ reads generated from Mi-msp-1 and Mi-msp-20 silenced second-stage juveniles (J2s) along with freshly hatched J2s were mapped to an already annotated M. incognita proteome to understand the impact on various nematode pathways. As compared to control, silencing of Mi-msp-1 caused differential expression of 29 transcripts, while Mi-msp-20 silencing resulted in differential expression of a broader set of 409 transcripts. In the Mi-msp-1 silenced J2s, cytoplasm (GO:0005737) was the most enriched gene ontology (GO) term, whereas in the Mi-msp-20 silenced worms, embryo development (GO:0009792), reproduction (GO:0000003) and nematode larval development (GO:0002119) were the most enriched terms. Limited crosstalk was observed between these two effectors as a sheer 5.9% of the up-regulated transcripts were common between Mi-msp-1 and Mi-msp-20 silenced nematodes. Our results suggest that in addition to the direct knock-down caused by silencing of Mi-msp-1 and Mi-msp-20, the cascading effect on other genes might also be contributing to a reduction in nematode's parasitic abilities.

Photorhabdus akhurstii is an insect–parasitic bacterium that symbiotically associates with the nematode, Heterorhabditis indica. The bacterium possesses several pathogenicity islands that aids in conferring toxicity to different insects. Herein, we constructed the plasmid clones of coding sequences of four toxin genes (pirA, tcaA, tccA and tccC; each was isolated from four P. akhurstii strains IARI-SGMG3, IARI-SGGJ2, IARI-SGHR2 and IARI-SGMS1) in Escherichia coli and subsequently, their biological activity were investigated against the fourth-instar larvae of the model insect, Galleria mellonella via intra-hemocoel injection. Bioinformatics analyses indicated inter-strain amino acid sequence difference at several positions of the candidate toxins. In corroboration, differential insecticidal activity of the identical toxin protein (PirA, TcaA, TccA and TccC conferred 15–59, 27–100, 25–100 and 33–98% insect mortality, respectively, across the strains) derived from the different bacterial strains was observed, suggesting that the diverse gene pool in Indian strains of P. akhurstii leads to strain-specific virulence in this bacterium. These toxin candidates appear to be an attractive option to deploy them in biopesticide development for managing the insect pests globally. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s13205-020-02288-0) contains supplementary material, which is available to authorized users.

Insulin-like growth factor 2 mRNA-binding protein 1 (IGF2BP1) is an mRNA-binding protein that has an oncofetal pattern of expression. It is also expressed in intestinal tissue, suggesting that it has a possible role in intestinal homeostasis. To investigate this possibility, here we generated Villin CreERT2:Igf2bp1flox/flox mice, which enabled induction of an IGF2BP1 knockout specifically in intestinal epithelial cells (IECs) of adult mice. Using gut barrier and epithelial permeability assays and several biochemical approaches, we found that IGF2BP1 ablation in the adult intestinal epithelium causes mild active colitis and mild-to-moderate active enteritis. Moreover, the IGF2BP1 deletion aggravated dextran sodium sulfate–induced colitis. We also found that IGF2BP1 removal compromises barrier function of the intestinal epithelium, resulting from altered protein expression at tight junctions. Mechanistically, IGF2BP1 interacted with the mRNA of the tight-junction protein occludin (Ocln), stabilizing Ocln mRNA and inducing expression of occludin in IECs. Furthermore, ectopic occludin expression in IGF2BP1-knockdown cells restored barrier function. We conclude that IGF2BP1-dependent regulation of occludin expression is an important mechanism in intestinal barrier function maintenance and in the prevention of colitis.

Owing to their health benefits, dietary fermentable fibers, such as refined inulin, are increasingly fortified in processed foods to enhance their nutritional value. However, we previously demonstrated that when inulin was fed to Toll-like receptor 5 deficient (T5KO) mice susceptible to dysbiosis, a subset of them developed cholestasis and subsequently liver cancer in a gut microbiota-dependent manner. Therefore, we hypothesized that clearance of bacterial taxa, and thereby gut metabolites, involved in the onset and progression to liver cancer could abate the disease in these mice. Such a reshaping of microbiota by vancomycin treatment was sufficient to halt the development of liver cancer in inulin-fed T5KO mice; however, this intervention did not remedy disease penetrance for cholestatic liver injury and its sequelae, including hyperbilirubinemia, hypolipidemia, cholemia and liver fibrosis. Selective depletion of gut bacterial communities was observed in vancomycin-treated mice, including Gram-positive Lachnospiraceae and Ruminococcaceae belonging to the phylum Firmicutes, Bifidobacteria of the phylum Actinobacteria, which ferment fibers, and Clostridium cluster XIVa, which produce secondary bile acids. Lack of liver cancer in vancomycin-treated mice strongly correlated with the substantial loss of secondary bile acids in circulation. Although cholemia was unabated by vancomycin, the composition of serum bile acids shifted toward an abundance of hydrophilic primary bile acids, denoted by the increase in conjugated-to-unconjugated bile acid ratio. Taken together, the present study suggests that microbiotal regulation of bile acid metabolism is one of the critical mediators of fermentable fiber-induced liver cancer in dysbiotic mice.

Magnetic nanoparticles with silica/gold (Au) shell were synthesized to be used as substrates for enzyme immobilization. Successful functionalization was followed by addition of phosphate linkers containing carboxyl end as free end for linking with enzyme. Effect of varied phospho-carboxyl chain lengths based linkers, 3-phosphonopropionic acid (3-PPA) and 16-phosphonopropionic acid (16-PHDA) on kinetic parameters of enzyme were studied. Immobilization of upto 670μg/mg of nanoparticles were observed. Enzyme bound with longer chain linker were observed to have Km (11.8mg/mL) and Vmax (1.59μmol/min/mg) which was found to be lower than 3-PPA. Reusability assay gave a minimum of 70% activity at 10th time of usage. Thus, an efficient, robust, cost effective and easily separable substrate for restraining the enzyme was prepared.

Lipocalin 2 (Lcn2) is a multifunctional innate immune protein that limits microbial overgrowth. Our previous study demonstrated that the gut microbiota directly induces intestinal Lcn2 production, and Lcn2-deficient ( Lcn2−/−) mice exhibit gut dysbiosis. Coincidentally, gut dysbiosis is associated with metabolic syndrome pathogenesis, and elevated Lcn2 levels has been considered a potential clinical biomarker of metabolic syndrome. Yet whether Lcn2 mitigates or exacerbates metabolic syndrome remains inconclusive. Our objective was to determine whether Lcn2 deficiency-induced compositional changes in gut microbiota contribute to gain in adiposity in aged mice. Utilizing Lcn2−/− mice and their wild-type (WT) littermates, we measured metabolic markers, including fasting blood glucose, serum lipids, fat pad weight, and insulin resistance at ages 3, 6, and 9 mo old. Relative to WT mice, aged Lcn2−/− mice exhibited a gain in adiposity associated with numerous features of metabolic syndrome, including insulin resistance and dyslipidemia. Surprisingly, supplementation with a high-fat diet did not further aggravate metabolic syndrome that spontaneously occurs in Lcn2−/− mice by 6 mo of age. Interestingly, chow-fed Lcn2−/− mice displayed marked differences in the bacterial abundance and metabolomic profile of the gut microbiota compared with WT mice. Overall, our results demonstrate that Lcn2 is essential to maintain metabolic and gut microbiotal homeostasis, where deficiency induces spontaneous delayed onset of metabolic syndrome.

In the efforts to develop a biocompatible transition metal complex as a drug alike for some of the prevailing non-communicable diseases (NCDs) and communicable diseases (CDs), a novel binuclear NiII compound [{NiII(hpdbal-sbdt)}2] (2) has been synthesized by the reaction of Ni(OAc)2.4H2O and H2hpdbal-sbdt (1) [1 is a dibasic tridentate ONS2− donor Schiff base ligand obtained by the condensation of 2-hydroxy-5-(phenyldiazenyl)benzaldehyde (Hhpdbal) and S-benzyldithiocarbazate (Hsbdt)]. Both ligand 1 and compound 2 were structurally characterized in the solid and solution state using various spectroscopic techniques like ATIR, 1H NMR, 13C NMR, TGA, FESEM, EDS and CHNS analysis. The antidiabetic activity of H2hpdbal-sbdt (1) and [{NiII(hpdbal-sbdt)}2] (2) were assessed using 2-NBDG uptake assay. The assay results showed 85% and 95% of fluorescent glucose uptake by insulin resistant HePG2 cells treated with compounds 1 and 2 respectively. The 2-NBDG uptake by the cells treated with the compound 2 was observed to be comparable to the standard antidiabetic drug metformin. Compounds 1 and 2 were also tested against five bacterial and two fungi strains in order to evaluate pathogen killing activity. Compound 2 showed significant inhibitory action towards the methicillin-resistant Staphylococcus aureus (MRSA) strain with an MIC value of 2 μg/mL whereas the ligand 1 was found to be inactive. Furthermore, the interactive nature of compound 2 with a model serum carrier protein bovine serum albumin (BSA) was studied using a multi-spectroscopic approach which provided an insight into the nature and extent of binding, conformational changes and the quenching of amino acid residues of the protein.

Root-knot nematodes are devastating pathogens of crop plants. The draft genome of southern root-knot nematode Meloidogyne incognita was published in 2008 and additional genome and transcriptome data became available later on. However, lack of a publically available annotation for M. incognita genome and transcriptome(s) limits the use of this data for functional and comparative genomics by the interested researchers. Here we present a comprehensive annotation for the M. incognita proteome data available at INRA Meloidogyne Genomic Resources page (https://meloidogyne.inra.fr/Downloads/Meloidogyne-incognita-V2-2017) and European Nucleotide Archive (ENA) (accession number: ERP009887) using a multi-pronged approach.

Insect-parasitic nematodes of the genus Heterorhabditis live in a symbiotic relationship with a gram negative Gamma-proteobacteria of the genus Photorhabdus. This nematode-bacteria pair is a simple and genetically tractable model to study animal-microbe symbiosis. Here we investigated the role of Heterorhabditis nematode C-type lectin (clec) genes in context of nematode-bacteria symbiosis. The in silico analysis identified seven clec genes in H. bacteriophora and three clec genes in H. indica. Two of the clec genes, H. bacteriophora clec-1 (Hb-clec-1) and H. bacteriophora clec-78 (Hb-clec-78) were further characterized. Both of these genes were present in a single copy in the H. bacteriophora genome. The phylogenetic analysis revealed that H. bacteriophora CLEC proteins were close to CLEC-1 and CLEC-78 proteins of free living Caenorhabditis but not to the CLEC proteins of insect-parasitic Steinernema nematodes which share a similar symbiotic relationship with Xenorhabdus bacteria. In situ hybridization showed that expression of Hb-clec-1 and Hb-clec-78 was localized to the alimentary canal of infective juveniles (IJs) in the region of terminal bulb, oesophago-intestinal valve and anterior part of intestine. Hb-clec-78 gene expression displayed significant positive correlation to the presence of bacteria during various stages of symbiosis: it was up-regulated during all the nematode developmental stages when Photorhabdus was symbiotically associated, but down-regulated at the post-IJ recovery stage when the developing nematodes were free of bacteria. Hb-clec-1 gene expression did not show any correlation with presence or absence of symbiont bacteria. Subject to genetic validation, our study suggests that Hb-clec-78 might be actively involved in modulation of symbiosis with Photorhabdus symbionts.

The present study is focused on assessment of phytoremediation potential of selected plant species by removal of endosulfan from contaminated soil via plant uptake. Eight plant species were selected for pot experiment under controlled condition. From the field monitoring study, Vetiveria zizanioides was found to be accumulate more endosulfan as compared to others. In this experiment, the phytoremediation potential of V. zizanioides is further tested. Apart from V. zizanioides, eight locally available plant species, namely Phragmitis karka, Jatropha curcas, Brassica juncea, Vigna radiata, Solanum lycopersicum, Solanum melongena, Spinacia oleracea and Withania somnifera, were also tested against different concentrations of endosulfan (0–1500 μg g−1) the interval of at 500 μg g−1. Morphological parameters in terms of fresh weight, biomass, shoot length and root length were deliberate just after harvesting. The chlorophyll, carotenoids and lipid peroxidation were estimated in plant samples. Microbial biomass carbon (MBC), dehydrogenase activity (DHA), pH, electrical conductivity and endosulfan concentration were analyzed in soil before and after cropping. The result shows that there was a noteworthy difference at 95% confidence level in growth of experimental plants when compared with control. Enhanced MBC and DHA showed active degradation of endosulfan by microbes that proliferate due to secretion of root exudates of test plants. Among all the test plants, V. zizanioides accumulated the highest and B. juncea accumulated the lowest concentration of endosulfan in their tissues. No significant reduction in lipid peroxidation and chlorophyll content in V. zizanioides supports its suitability for phytoremediation.

Ethnopharmacological relevance Xylocarpus moluccensis (Lamk.) M. Roem of family Meliaceae has triterpenoids rich fruits. Triterpenoids have been known to possess cardioprotection and anti-atherosclerotic activities (Han and Bakovic, 2015; Wu et al., 2009). Standardized fraction of these fruits exhibited anti-dyslipidemic (Srivastava et al., 2015), anti-inflammatory (Ravangpai et al., 2011) and CNS depressant activity (Sarker et al., 2007). However, there is no report in the literature on its cardiovascular effects. Aim of the study The present study was undertaken to assess vasoprotective, anti-atherosclerotic and further examine the anti-dyslipidemic effect of the standardized fraction of Xylocarpus moluccensis (F018) fruits in the mechanical injury and high fat diet (HFD) induced dyslipidemic/ atherosclerosis models. Materials and methods Guinea pigs were fed 0.08% cholesterol + 15% fat diet for 3 weeks, while ApoE KO mice were fed high fat diet for 18 weeks to induce dyslipidemia and atherosclerosis. A combination of balloon injury and high fat diet (1% cholesterol, 6% peanut oil) for 5 weeks was used to accelerate atherosclerosis in NZW rabbits. F018 was administered once daily by oral route in guinea pigs (10, 25 or 50 mg/kg/day for 3 weeks), ApoE KO mice (50 mg/kg/day for 6 weeks) and in NZW rabbit (25 mg/kg/day for 5 weeks) to monitor its effect on dyslipidemia, vasoreactivity and plaque composition by using standard methodologies. Results F018 treatment in guinea pigs (25 and 50 mg/kg/day), ApoE mice (50 mg/kg/day) and rabbits (25 mg/kg/day) significantly reduced plasma lipids and improved ACh induced vasorelaxation. Anti-dyslipidemic effect of F018 seems to be due to the modulation of enterohepatic genes involved in the cholesterol absorption and excretion. Moreover, significant improvement in the acetylcholine (ACh) induced vasorelaxation was accompanied with reduced inflammatory burden and enhanced activation of eNOS in ApoE mice aortic tissue. Similarly inflammatory cytokines, immunolabeling of macrophage marker (CD68) and MMP-9 were reduced along with augmentation in vascular smooth muscle cells and collagen type I and III in the mechanically injured iliac artery segment in the rabbits. Conclusions Altogether, F018 preserved vasoreactivity, reduced atherosclerotic plaque progression and enhanced plaque stability by reducing lipids, inflammatory cytokines, improving endothelial function and collagen content.

The rice root-knot nematode Meloidogyne graminicola has emerged as a devastating pest of rice in South-East Asian countries. Here we present a draft genome sequence for M. graminicola, assembled using data from short and long insert libraries sequenced on Illumina GAIIx sequencing platform.

India, with around 15 million COVID-19 cases, recently became the second worst-hit nation by the SARS-CoV-2 pandemic. In this study, we analyzed the mutation and selection landscape of 516 unique and complete genomes of SARS-CoV-2 isolates from India in a 12-month span (from Jan to Dec 2020) to understand how the virus is evolving in this geographical region. We identified 953 genome-wide loci displaying single nucleotide polymorphism (SNP) and the Principal Component Analysis and mutation plots of the datasets indicate an increase in genetic variance with time. The 42% of the polymorphic sites display substitutions in the third nucleotide position of codons indicating that non-synonymous substitutions are more prevalent. These isolates displayed strong evidence of purifying selection in ORF1ab, spike, nucleocapsid, and membrane glycoprotein. We also find some evidence of localized positive selections ORF1ab, spike glycoprotein, and nucleocapsid. The CDSs for ORF3a, ORF8, nucleocapsid phosphoprotein, and spike glycoprotein were found to evolve at rapid rate. This study will be helpful in understanding the dynamics of rapidly evolving SARS-CoV-2., Graphical Abstract ga1

Plant-parasitic nematodes (PPNs) cause substantial damage to agricultural and horticultural crops around the world. The early, accurate and rapid detection of PPNs at field level is the most important step to manage these pests, but remains a challenge for researchers. Loop-mediated isothermal amplification (LAMP) has emerged as a robust, rapid and sensitive method for point-of-service detection and diagnosis of plant pathogens such as bacteria, fungus, virus and nematodes. The present review summarises and analyses the LAMP assays developed for the detection and diagnosis of various important PPNs. Different gene targets such as SSU, 5.8S, LSU, ITS-1, ITS-2, D2-D3, IGS-1 and IGS-2 regions of the rRNA, syg-2 and pel-3 genes have been used for the development of the highly sensitive, species-specific or genus-specific LAMP assays for various PPNs. The LAMP assays are superior to PCR based assays as they are faster, more sensitive and can detect nematode DNA in up to femtogram quantities from the soil, plant tissue, preserved nematode samples or nematode genomic DNA stabilised on cellulose cards. The results of the LAMP assays can be visualised in the reaction tube by DNA intercalating dyes, observation of sample turbidity, by gel electrophoresis, and through lateral flow assays. Presently the LAMP-based assays are gaining popularity as the method of choice for point-of-care detection and diagnosis of various PPNs.

Mammalian siderophores are believed to play a critical role in maintaining iron homeostasis. However, the properties and functions of mammalian siderophores have not been fully clarified. In this study, we have employed Chrome Azurol S (CAS) assay which is a well-established method for bacterial siderophores study, to detect and quantify mammalian siderophores in urine samples. Our study demonstrates that siderophores in urine can be altered by diet, gut microbiota and inflammation. C57BL/6 mice, fed on plant-based chow diets which contain numerous phytochemicals, have more siderophores in the urine compared to those fed on purified diets. Urinary siderophores were up-regulated in iron overload conditions, but not altered by other tested nutrients status. Further, germ-free mice displayed 50% reduced urinary siderophores, in comparison to conventional mice, indicating microbiota biotransformation is critical in generating or stimulating host metabolism to create more siderophores. Altered urinary siderophores levels during inflammation suggest that host health conditions influence systemic siderophores level. This is the first report to measure urinary siderophores as a whole, describing how siderophores levels are modulated under different physiological conditions. We believe that our study opens up a new field in mammalian siderophores research and the technique we used in a novel manner has the potential to be applied to clinical purpose.

// Laura Pellegrini 1 , Jiaming Xue 1, 3 , David Larson 1 , Sandra Pastorino 1 , Sandro Jube 2 , Kelly H. Forest 3 , Zeyana Salim Saad-Jube 4 , Andrea Napolitano 1, 5 , Ian Pagano 1 , Vishal S. Negi 5 , Marco E. Bianchi 6 , Paul Morris 7 , Harvey I. Pass 8 , Giovanni Gaudino 1 , Michele Carbone 1 , Haining Yang 1 1 University of Hawai'i Cancer Center, University of Hawai'i at Manoa, Honolulu, HI, USA 2 University of Hawai'i at Manoa, Department of Cell and Molecular Biology, John A. Burns School of Medicine, Honolulu, HI, USA 3 Leeward Community College, Mathematics and Sciences Division, University of Hawai'i System, Pearl City, HI, USA 4 University of Hawai'i at Manoa, Myron B. Thompson School of Social Work, Office of Public Health and Center on Aging, Honolulu, HI, USA 5 University of Hawai'i at Manoa, Department of Molecular Biosciences and Bioengineering, Honolulu, HI, USA 6 San Raffaele University and Research Institute, Milan, Italy 7 Department of Thoracic Surgery, Queen's Medical Center, Honolulu, HI, USA 8 New York University School of Medicine, Department of Cardiothoracic Surgery, New York, NY, USA Correspondence to: Haining Yang, email: hyang@cc.hawaii.edu Michele Carbone, email: mcarbone@cc.hawaii.edu Keywords: HMGB1, RAGE, ethyl pyruvate, mesothelioma, therapeutic Received: June 30, 2016 Accepted: January 23, 2017 Published: February 07, 2017 ABSTRACT Human malignant mesothelioma (MM) is an aggressive cancer linked to asbestos and erionite exposure. We previously reported that High-Mobility Group Box-1 protein (HMGB1), a prototypic damage-associated molecular pattern, drives MM development and sustains MM progression. Moreover, we demonstrated that targeting HMGB1 inhibited MM cell growth and motility in vitro , reduced tumor growth in vivo , and prolonged survival of MM-bearing mice. Ethyl pyruvate (EP), the ethyl ester of pyruvic acid, has been shown to be an effective HMGB1 inhibitor in inflammation-related diseases and several cancers. Here, we studied the effect of EP on the malignant phenotype of MM cells in tissue culture and on tumor growth in vivo using an orthotopic MM xenograft model. We found that EP impairs HMGB1 secretion by MM cells leading to reduced RAGE expression and NF-κB activation. As a consequence, EP impaired cell motility, cell proliferation, and anchorage-independent growth of MM cells. Moreover, EP reduced HMGB1 serum levels in mice and inhibited the growth of MM xenografts. Our results indicate that EP effectively hampers the malignant phenotype of MM, offering a novel potential therapeutic approach to patients afflicted with this dismal disease.