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Nanoparticles serve various purposes in the hydrocarbon industry which are utilized to improve the volume of oil and gas production. Nanomaterials are used in petroleum applications as nanoparticles, nanosensors, nanocomposites, coated nanoparticles, nanofluids etc. Thus, the application of nanotechnology in the enhanced oil recovery (EOR) method helps to extract the trapped oil present beneath the surface. Some nano ultra-fine particles are cost-effective and eco-friendly. The injected fluid interacts with the rock/oil system to create a favourable environment for oil recovery. This study focuses on a review of the different types of nanoparticles and nanomaterials used in petroleum applications. This paper aims to study the classification and characterization of nanoparticles. This also includes the physiochemical and mechanical properties of nanomaterials. The study discusses the applications of nanoparticles in chemical, miscible, thermal and microbial floodings. It mainly focuses on various mechanisms involved by adding nanoparticles such as wettability alteration, IFT reduction, rheology improvement, mobility control etc. This research also addresses new green nanomaterials and nanocomposites, which includes injecting specialized green chemicals (surfactants, alcohols, and polymers) that successfully displace oil. The IFT between the displacing fluid and the oil is reduced due to phase-behaviour properties.

Hepcidin, a circulatory hepatic peptide hormone, is associated with systemic iron homeostasis. Inflammation leads to an increase in hepcidin expression, which dysregulates body iron level. The related disorder, anemia of inflammation, is the second most prevalent anemia-related disorder worldwide. In the present study, we conducted in vitro and in vivo studies to evaluate the effect of black pepper (BP) and its major bioactive alkaloid, piperine, on anemia of inflammation. The initial in vitro study using human hepatocyte cell line, HepG2, confirmed that among different black pepper extracts: methanol (BPME), ethanol (BPEE) and aqueous (BPAE), BPME to be most effective in downregulating transcription of hepcidin gene. Further, BPME and piperine significantly downregulated hepcidin protein expression at 200 μg/ml and 100 μM concentrations, respectively. In the next phase, BPME and piperine were found to significantly attenuate BMP-6 and IL-6 induced hepcidin overexpression by downregulating the increased level of pSMAD1 and pSTAT3 proteins, respectively. For in vivo study, we first subcutaneously injected male BALB/c mice with oil of turpentine, thrice within a period of two weeks, in order to enhance the expression of hepcidin. After that, the intraperitoneal administration of BPME and piperine at 70 and 25 mg/kg body weight, respectively, on alternate days for a period of another two weeks resulted in downregulation of hepcidin overexpression in diseased mice, as confirmed by RT-PCR and immunoblot analysis. The histopathology of liver tissue confirmed increased iron bioavailability in BPME and piperine treated animals. The molecular docking-based interaction studies demonstrated the binding potential of piperine with SMAD1 and STAT3 proteins. The binding patterns supported the proposed inhibition of hepcidin activating proteins. All together, these findings suggest black pepper as a therapeutic candidate for the treatment of anemia of inflammation.

Main aim of the present research is to explore the potential use of Vetiveria zizanioides L. for phytoremediation of arsenic, fluoride, and manganese simultaneously from synthetic wastewater in a batch scale floating platform unit. Half strength Hoagland's nutrient solution spiked with arsenic, fluoride, and manganese concentrations of 1, 20, and 10 mg/L, respectively has been used. The effects of pH and treatment time on simultaneous removal of arsenic, fluoride, and manganese have been performed. V. zizanioides has exhibited optimum growth at pH 8 and the removal of arsenic and fluoride is observed to be 59.6 and 38.1%, respectively. This plant has successfully removed all of the manganese (99.3%). The uptake of manganese is found to be faster than the arsenic and fluoride. The trend of arsenic, fluoride, and manganese accumulation in various parts of V. zizanioides is found as roots > stems > leaves. Result showed that the use of V. zizanioides would be appropriate to treat arsenic, fluoride, and manganese contaminated wastewater.

Papaya fruits have great nutritional and economical values. The ripening stage of papaya fruits at the time of harvesting massively impacts the quality and shelf-life of fruits. Volatile organic compounds (VOCs) emitting from the papaya fruits during the ripening process could be used as a real-time non-invasive biomarker to characterize the ripening stage. This paper reports identification of VOCs as non-invasive ripening biomarkers of papaya fruits based on solid-phase micro-extraction coupled with gas chromatography–mass spectrometry (GC–MS) analysis of VOCs emitted from the papaya fruit cv. ‘Red Lady’. Three ripening stages were studied, viz., green unripe (UR), yellowish-green intermediate ripe (IR), and yellow full ripe (FR). GC–MS analyses and the subsequent statistical studies identified a total of 35 VOCs. Partial Least Squares Discriminant Analysis (PLS-DA) of VOCs from the three ripening stages identified six biomarker VOCs, which can efficiently distinguish between ripening stages. Among the six ripening biomarkers, three VOCs (methyl hexanoate, 3-carene and longifolene) showed a remarkable correlation with the ripening-associated changes in the fruit nutritional profile. The biomarkers reported here could be used as a viable technology for non-invasive monitoring of ripening stages and the nutritional value of papaya fruits.

Graphical abstract, Highlights • Wheatgrass extract inhibits production of nitric oxide and reactive oxygen species. • Wheatgrass extract inhibits production of pro-inflammatory and inflammatory markers. • Molecular docking suggests inhibition of COX-2 by major metabolites of wheatgrass., Inflammation is a multifaceted set of cellular communications generated against foreign infection, toxic influence or autoimmune injury. The present study investigates the anti-inflammatory effect of wheatgrass extract against the harmful impact of lipopolysaccharide (LPS) in macrophage cells, i.e., RAW 264.7 cells. Our results indicate that 5- and 7- days old wheatgrass extracts inhibit the LPS-stimulated production of nitric oxide. Moreover, wheatgrass extract significantly downregulates the mRNA expression of LPS-stimulated various pro-inflammatory markers, tumor necrosis factor-α, interleukin-6, interleukin-1β, AP-1 and also iNOS-2 and COX-2. Our flow cytometry analyses confirmed that wheatgrass extract prevents the generation of reactive oxygen species in LPS-stimulated RAW 264.7 cells, thus arresting oxidative stress in cells. The immunoblot analyses also confirmed a significant reduction in the expression of inflammatory proteins, namely, iNOS-2 and COX-2, in wheatgrass extract-treated cells, compared to LPS-stimulated condition. The NF-κB transactivation assay further confirmed the inhibitory effect of wheatgrass extracts on the LPS-stimulated expression of NF-κB. Molecular docking based studies showed the plausible binding of two significant wheatgrass constituents, i.e., apigenin and myo-inositol with COX-2 protein, with binding energies of −10.59 kcal/mol and −7.88 kcal/mol, respectively. Based on the above results, wheatgrass may be considered as a potential therapeutic candidate for preventing inflammation.

Interaction of SARS-CoV-2 spike glycoprotein with the ACE2 cell receptor is very crucial for virus attachment to human cells. Selected mutations in SARS-CoV-2 S-protein are reported to strengthen its binding affinity to mammalian ACE2. The N501T mutation in SARS-CoV-2-CTD furnishes better support to hotspot 353 in comparison with SARS-CoV and shows higher affinity for receptor binding. Recombination analysis exhibited higher recombination events in SARS-CoV-2 strains, irrespective of their geographical origin or hosts. Investigation further supports a common origin among SARS-CoV-2 and its predecessors, SARS-CoV and bat-SARS-like-CoV. The recombination events suggest a constant exchange of genetic material among the co-infecting viruses in possible reservoirs and human hosts before SARS-CoV-2 emerged. Furthermore, a comprehensive analysis of codon usage bias (CUB) in SARS-CoV-2 revealed significant CUB among the S-genes of different beta-coronaviruses governed majorly by natural selection and mutation pressure. Various indices of codon usage of S-genes helped in quantifying its adaptability in other animal hosts. These findings might help in identifying potential experimental animal models for investigating pathogenicity for drugs and vaccine development experiments.

Osteosarcoma is the most frequently occurring cancer in children as well as young adolescents and the metastatic forms worsen this condition to a further great extent. The metastatic dissemination of cancer cells is often acquired through a process of epithelial-mesenchymal transition (EMT). Since, phytochemicals have attracted intense interest in recent years due to their diverse pharmacological effects, in the present study, we investigated if berberine, a naturally occurring isoquinoline quaternary alkaloid, could modulate the EMT in osteosarcoma cells. Our experimental studies showed that berberine reduced cell viability, colony formation, wound healing ability and migration of osteosarcoma cells. Also, berberine significantly reduced the expression of matrix metalloproteinase (MMP)-2, suggesting its inhibitory action on the matrix metalloproteinases that are required for cancer cell invasion. The significant reduction in the expression of vimentin, N-cadherin, fibronectin and increased expression of E-cadherin further suggested its role in the inhibition of EMT in osteosarcoma cells. The downregulation of H3K27me3, as well as the decreased expression of the histone methyl transferase enzyme EZH2, further substantiated the fact that the plant alkaloid can be used as an epigenetic modulator in the treatment of osteosarcoma. In conclusion, our findings suggest that berberine inhibits proliferation and migration of osteosarcoma cells and most importantly reverses EMT along with modulation of key epigenetic regulators.

Zinc is essential for growth and development of all living organisms, especially human being. Deficiency of micronutrients like zinc and iron has been linked to the manifestation of hidden hunger. Therefore, it is imperative that development of some rapid screening method for bioavailable zinc in various crops and food commodities would be an essential addition in battle against zinc deficiency related hidden hunger. One such method could be the usage of fluorescence based zinc ion sensing probe which would be robust and convenient for estimating bioavailable zinc. To address this issue, NBD-TPEA, a highly sensitive zinc ion sensing probe, have been used in this study towards the development of a novel fluorescence based approach for the analysis of zinc bioavailability in Caco-2 cells as an in-vitro cellular model. The use of this probe showed dose dependent sensitivity towards increasing concentrations of zinc ion uptake by Caco-2 cells. It also showed specificity for zinc ion uptake as compared to other metal ions in-vitro. These observations correlated extremely well with zinc uptake analysis by cell imaging and conventional analytical technique like, ICP-MS. The developed assay was then tested in mushroom and some selected biofortified derivatives of wheat for determining the levels of their bioavailable zinc using Caco-2 cells. The data as obtained with these food samples in our developed bioassay correlated well with the other sophisticated analytical techniques thus validating our cell based assay. Hence, the developed assay could serve as a simple but sensitive tool for determining bioavailable zinc in various food samples.

Diffuse large B-cell lymphoma (DLBCL) is the most common subtype of non-Hodgkin lymphoma; 40% of patients relapse after a complete response or are refractory to therapy. To survive, the activated B-cell (ABC) subtype of DLBCL relies upon B-cell receptor signaling, which can be modulated by the activity of Bruton tyrosine kinase (BTK). Targeting BTK with ibrutinib, an inhibitor, provides a therapeutic approach for this subtype of DLBCL. However, non-Hodgkin lymphoma is often resistant to ibrutinib or acquires resistance soon after exposure. We explored how this resistance develops. We generated 3 isogenic ibrutinib-resistant DLBCL cell lines and investigated the deregulated pathways known to be associated with tumorigenic properties. Reduced levels of BTK and enhanced phosphatidylinositol 3-kinase (PI3K)/AKT signaling were hallmarks of these ibrutinib-resistant cells. Upregulation of PI3K-β expression was demonstrated to drive resistance in ibrutinib-resistant cells, and resistance was reversed by the blocking activity of PI3K-β/δ. Treatment with the selective PI3K-β/δ dual inhibitor KA2237 reduced both tumorigenic properties and survival-based PI3K/AKT/mTOR signaling of these ibrutinib-resistant cells. In addition, combining KA2237 with currently available chemotherapeutic agents synergistically inhibited metabolic growth. This study elucidates the compensatory upregulated PI3K/AKT axis that emerges in ibrutinib-resistant cells.

The Cambay basin of Western India has been proved as the most prominent and promising basin for major shale oil and gas discoveries. The present work is an exploration study of shales of different tectonic blocks of Cambay basin, which characterizes it as potential source as well as a reservoir for hydrocarbon generation and expulsion. Major experimental work was done on the Cambay shale of North tectonic block of Cambay basin and it was found that the shale has good to excellent organic richness, hybrid oil/gas bearing potential, mature shale potential, good hydrocarbon generation potential and good kerogen transformation. On the contrary, shales of south tectonic block are at the immature phase of hydrocarbon generation and may generate hydrocarbons upon thermal maturation and deep burial. The performed analysis and interpretation are explaining the true hydrocarbon potential of shales of Cambay basin and hence providing with a roadmap for the shale geochemical characterization which may be useful for development of shale plays at large scale.

Water is often considered as the highest performance working fluid for liquid–vapor phase change due to its high thermal conductivity and large enthalpy of vaporization. However, a wide range of industrial systems require using low surface tension liquids where heat transfer enhancement has proved challenging for boiling and evaporation. Here, we enable a new paradigm of phase change heat transfer, which favors high volatility, low surface tension liquids rather than water. We utilized a nanoporous membrane of ≈600 nm thickness and

The intracellular concentration of reduced glutathione (GSH) lies in the range of 1–10 mM, thereby indisputably making it the most abundant intracellular thiol. Such a copious amount of GSH makes it the most potent and robust cellular antioxidant that plays a crucial role in cellular defence against redox stress. The role of GSH as a denitrosylating agent is well established; in this study, we demonstrate GSH mediated denitrosylation of HepG2 cell-derived protein nitrosothiols (PSNOs), by a unique spin-trapping mechanism, using 5,5-dimethyl-1-pyrroline N-oxide (DMPO) as the spin trapping agent, followed by a western blot analysis. We also report our findings of two, hitherto unidentified substrates of GSH mediated S-denitrosylation, namely S-nitrosoglutaredoxin 1 (Grx1-SNO) and S-nitrosylated R1 subunit of ribonucleotide reductase (R1-SNO).

Phthalate, a plasticizer, endocrine disruptor, and potential carcinogen, is degraded by a variety of bacteria. This degradation is initiated by phthalate dioxygenase (PDO), a Rieske oxygenase (RO) that catalyzes the dihydroxylation of phthalate to a dihydrodiol. PDO has long served as a model for understanding ROs despite a lack of structural data. Here we purified PDOKF1 from Comamonas testosteroni KF1 and found that it had an apparent kcat/Km for phthalate of 0.58 ± 0.09 μM−1s−1, over 25-fold greater than for terephthalate. The crystal structure of the enzyme at 2.1 A resolution revealed that it is a hexamer comprising two stacked α3 trimers, a configuration not previously observed in RO crystal structures. We show that within each trimer, the protomers adopt a head-to-tail configuration typical of ROs. The stacking of the trimers is stabilized by two extended helices, which make the catalytic domain of PDOKF1 larger than that of other characterized ROs. Complexes of PDOKF1 with phthalate and terephthalate revealed that Arg207 and Arg244, two residues on one face of the active site, position these substrates for regiospecific hydroxylation. Consistent with their roles as determinants of substrate specificity, substitution of either residue with alanine yielded variants that did not detectably turnover phthalate. Together, these results provide critical insights into a pollutant-degrading enzyme that has served as a paradigm for ROs and facilitate the engineering of this enzyme for bioremediation and biocatalytic applications.

A long-standing mystery in vertebrate Hedgehog signaling is how Patched 1 (PTCH1), the receptor for Hedgehog ligands, inhibits the activity of Smoothened, the protein that transmits the signal across the membrane. We previously proposed (Kinnebrew et al., 2019) that PTCH1 inhibits Smoothened by depleting accessible cholesterol from the ciliary membrane. Using a new imaging-based assay to directly measure the transport activity of PTCH1, we find that PTCH1 depletes accessible cholesterol from the outer leaflet of the plasma membrane. This transport activity is terminated by binding of Hedgehog ligands to PTCH1 or by dissipation of the transmembrane potassium gradient. These results point to the unexpected model that PTCH1 moves cholesterol from the outer to the inner leaflet of the membrane in exchange for potassium ion export in the opposite direction. Our study provides a plausible solution for how PTCH1 inhibits SMO by changing the organization of cholesterol in membranes and establishes a general framework for studying how proteins change cholesterol accessibility to regulate membrane-dependent processes in cells. Ministry of Education (MOE) Published version Christian Siebold - Cancer Research UK grant reference numbers C20724 and A26752, European Research Council grant reference number 647278. Rajat Rohatgi - National Institutes of Health grant reference numbers GM118082 and GM106078. Arun Radhakrishnan - National Institutes of Health grant reference number HL20948, Welch Foundation grant reference number I-1793 and Leducq Foundation grant reference number 19CVD04. Yasunori Saheki - Ministry of Education, Singapore grant reference numbers MOE2017-T2-2-001 and MOE-T2EP30120-0002. Maia Kinnebrew - National Science Foundation Predoctoral Fellowship. Giovanni Luchetti - Ford Foundation Predoctoral Fellowship.

Plants respond to environmental cues via adaptive cell reprogramming that can affect whole plant and ecosystem functionality. Microbiota constitutes part of the inner and outer environment of the plant. This Umwelt underlies steady dynamics, due to complex local and global biotic and abiotic changes. Hence, adaptive plant holobiont responses are crucial for continuous metabolic adjustment at the systems level. Plants require oxygen-dependent respiration for energy-dependent adaptive morphology, such as germination, root and shoot growth, and formation of adventitious, clonal, and reproductive organs, fruits, and seeds. Fermentative paths can help in acclimation and, to our view, the role of alternative oxidase (AOX) in coordinating complex metabolic and physiological adjustments is underestimated. Cellular levels of sucrose are an important sensor of environmental stress. We explored the role of exogenous sucrose and its interplay with AOX during early seed germination. We found that sucrose-dependent initiation of fermentation during the first 12 h after imbibition (HAI) was beneficial to germination. However, parallel upregulated AOX expression was essential to control negative effects by prolonged sucrose treatment. Early downregulated AOX activity until 12 HAI improved germination efficiency in the absence of sucrose but suppressed early germination in its presence. The results also suggest that seeds inoculated with arbuscular mycorrhizal fungi (AMF) can buffer sucrose stress during germination to restore normal respiration more efficiently. Following this approach, we propose a simple method to identify organic seeds and low-cost on-farm perspectives for early identifying disease tolerance, predicting plant holobiont behavior, and improving germination. Furthermore, the research strengthens the view that AOX can serve as a powerful functional marker source for seed hologenomes.

SummaryMicronutrient deficiency also termed hidden hunger affects a large segment of the human population, particularly in developing and underdeveloped nations. Tomato the second most consumed vegetable crop in the world after potato can serve as a sustainable source to alleviate micronutrient deficiency. In tomato, the mutations in the R2R3-MYB117 transcription factor elicit trifoliate leaves and initiate axillary meristems; however, its effect on fruit metabolome remains unexplored. The fruits of a new trifoliate (tf) allele (tf-5) were firmer, had higher °Brix, folate, and carotenoids. The transcriptome, proteome, and metabolome profiling of tf-5 reflected a broad-spectrum change in homeostasis. The tf-5 allele enhanced the fruit firmness by suppressing cell wall softening-related proteins. The tf-5 fruit displayed a substantial increase in aminome, particularly γ-aminobutyric acid, with a parallel reduction in aminoacyl t-RNA synthases. The increased lipoxygenases proteins and transcripts seemingly elevated jasmonic acid. In addition, increased abscisic acid hydrolases transcripts coupled with reduced precursor supply lowered abscisic acid. The upregulation of carotenoids was mediated by modulation of methylerythreitol and plastoquinone pathways along with an increase in carotenoids isomerization proteins. The upregulation of folate in tf-5 was connoted by the increase in precursor p-aminobenzoic acid and transcripts of several folate biosynthesis pathway genes. The reduction in pterin-6-carboxylate and γ-glutamyl hydrolase activity indicated that the diminished folate degradation also enriched folate levels. Our study delineates that introgression of the tf-5 can be used for the γ-aminobutyric acid, carotenoids, and folate fortification of tomato.One-sentence summaryA tomato trifoliate allele encoding a truncated MYB117 transcription factor alters cellular homeostasis and fortifies γ-aminobutyric acid, folate, and carotenoids in tomato fruits.

The rice root system is primarily composed of shoot-borne adventitious/crown roots (ARs/CRs) that develop from the coleoptile base, and therefore, it is an excellent model system for studying shoot-to-root trans-differentiation process. We reveal global changes in protein and metabolite abundance and protein phosphorylation in response to an auxin stimulus during CR development. The liquid chromatography–tandem mass spectrometry (LC–MS/MS) and gas chromatography–mass spectrometry (GC–MS) analyses of developing crown root primordia (CRP) and emerged CRs identified 334 proteins and 12 amino acids, respectively, that were differentially regulated upon auxin treatment. Gene ontology enrichment analysis of global proteome data uncovered the biological processes associated with chromatin conformational change, gene expression and cell cycle that were regulated by auxin signaling. Spatial gene expression pattern analysis of differentially abundant proteins disclosed their stage-specific dynamic expression pattern during CRP development. Further, our tempo-spatial gene expression and functional analyses revealed that auxin creates a regulatory module during CRP development and activates ethylene biosynthesis exclusively during CRP initiation. Further, the phosphoproteome analysis identified 8,220 phosphosites, which could be mapped to 1,594 phosphoproteins and of which 66 phosphosites were differentially phosphorylated upon auxin treatment. Importantly, we observed differential phosphorylation of the cyclin-dependent kinase G-2 (OsCDKG;2) and cell wall proteins, in response to auxin signaling, suggesting that auxin-dependent phosphorylation may be required for cell cycle activation and cell wall synthesis during root organogenesis. Thus, our study provides evidence for the translational and post-translational regulation during CR development downstream of the auxin signaling pathway.

Apple (Malus sp.) and other genera belonging to the sub-tribe Malinae of the Rosaceae family produce unique benzoic acid-derived biphenyl phytoalexins. Cell cultures of Malus domestica cv. 'Golden Delicious' accumulate two biphenyl phytoalexins, aucuparin and noraucuparin, in response to the addition of a Venturia inaequalis elicitor (VIE). In this study, we isolated and expressed a cinnamate-CoA ligase (CNL)-encoding sequence from VIE-treated cell cultures of cv. 'Golden Delicious' (M. domestica CNL; MdCNL). MdCNL catalyses the conversion of cinnamic acid into cinnamoyl-CoA, which is subsequently converted to biphenyls. MdCNL failed to accept benzoic acid as a substrate. When scab-resistant (cv. 'Shireen') and moderately scab-susceptible (cv. 'Golden Delicious') apple cultivars were challenged with the V. inaequalis scab fungus, an increase in MdCNL transcript levels was observed in internodal regions. The increase in MdCNL transcript levels could conceivably correlate with the pattern of accumulation of biphenyls. The C-terminal signal in the MdCNL protein directed its N-terminal reporter fusion to peroxisomes in Nicotiana benthamiana leaves. Thus, this report records the cloning and characterisation of a cinnamoyl-CoA-forming enzyme from apple via a series of in vivo and in vitro studies. Defining the key step of phytoalexin formation in apple provides a biotechnological tool for engineering elite cultivars with improved resistance.

An ideal scaffold which mimics extracellular matrix as an artificial bone graft substitute is the need of current clinical management of orthopaedic complications. In this study, a porous biomimetic scaffold was synthesized using in situ gas foaming method. Briefly, a blend of gelatin and polyvinylpyrrolidone (PVP) was crosslinked with glutaraldehyde and lyophilized. The blending of two polymers were confirmed with FTIR, XRD and TGA analysis. The synthesized scaffold was further characterized for its biocompatibility and osteogenic potential. The physico-chemical properties like microarchitecture, porosity, water adsorption ability, and mechanical strength were also investigated. Further, the enhanced proliferation and migration of murine mesenchymal stem cells (C3H10t1/2 cells) through interconnected pores of the scaffold over an extended time period suggests the cytocompatibility of scaffold. Next, the biocompatibility of the scaffold was confirmed by in ovo implantation on the chorioallantoic membrane (CAM) using chicken embryo. In addition, increased matrix mineralization was confirmed by alizarin red staining and EDX analysis of apatite depositions over the scaffold, when induced with osteogenic media. In summary, these findings demonstrate biocompatibility and osteo-inductive potentials of the gelatin-PVP biomimetic polymer composite scaffold with suitability as bone graft substitute material.

A simple, rapid and selective high-performance thin-layer chromatographic (HPTLC) method has been developed and validated for simultaneous determination of three withanolides (withaferin A, withanone and withanolide A) and three phenolic acids (caffeic acid, ferulic acid and benzoic acid) from different parts (root, stem and leaf) of Withania somnifera and its two commercially available polyherbal formulations. The extraction efficiency of withanolides and phenolic acids were tested using two solvents, chloroform and methanol, respectively. HPTLC separation was performed on silica coated aluminium plates Si 60F254; using toluene, ethyl acetate and acetic acid (60:40:4). The samples were quantitated at 231 nm. The purity and identity of peaks of all the six analytes were confirmed by matching Rf values and UV-spectrum with authentic standards. The identity of three withanolides was further confirmed by positive ion electrospray ionization mass spectrometry (ESI-MS/MS) analyses. The developed method was validated for sensitivity, linearity, reproducibility, accuracy, the limit of detection (LOD) and limit of quantification (LOQ) following the guidelines of the International Conference on Harmonization (ICH). The method was found to be linear (r > 0.99) in the range of 50–2000 ng/band for benzoic acid and 50–1000 ng/band for the other five studied metabolites. This simple and accurate HPTLC method provided enhanced resolution of studied analytes as compared to other phytoconstituents present in W. somnifera extracts. It has also been successfully applied in the analysis and quantification of two polyherbal formulations containing W. somnifera plant parts.

A novel solid acid catalyst was prepared from the palm-fruit-bunch by in-situ incomplete sulphonation carbonization method. The prepared catalyst was characterized to determine its physical and che...

Background: Atrial fibrillation is associated with increased risks of death, stroke/systemic embolism, and bleeding (incurred by antithrombotic therapy), which may occur early after diagnosis. Methods: We assessed the risk of early events (death, stroke/systemic embolism, and major bleeding) over 12 months and their relation to the time after diagnosis of atrial fibrillation in 52 014 patients prospectively enrolled in the GARFIELD-AF registry (Global Anticoagulant Registry in the FIELD–Atrial Fibrillation) between March 2010 and August 2016. Results: Over 12 months, 2140 patients died (mortality rate, 4.3; 95% CI, 4.2–4.5 per 100 person-years), of whom 288 (13.5%) died in the first month (6.8; 95% CI, 6.1–7.6). Over 12 months, 657 patients had a stroke/systemic embolism (1.3; 95% CI, 1.2–1.4) and 411 had a major bleeding (0.8; 95% CI, 0.8–0.9). During the first month, the rates (per 100 person-years) of stroke/systemic embolism and major bleed were 2.3 (95% CI, 1.9–2.8) and 1.5 (95% CI, 1.2–1.9), respectively. The elevated 1-month mortality rate was mostly attributable to cardiovascular mortality (3.5; 95% CI, 3.0–4.1), in particular, heart failure, sudden death, and acute coronary syndromes (1.0 [95% CI, 0.8–1.4], 0.6 [95% CI, 0.4–0.8], and 0.5 [95% CI, 0.3–0.8], respectively). Age, heart failure, prior stroke, history of cirrhosis, vascular disease, moderate-to-severe kidney disease, diabetes mellitus, and living in North or Latin America were independent predictors of a higher risk of early death, whereas anticoagulation and living in Europe or Asia were independent predictors of a lower risk of early death. A predictive model developed for the 1-month risk of death had a C-statistic of 0.81 (95% CI, 0.78–0.83). Conclusions: The increased hazard of early events, in particular, cardiovascular mortality, in newly diagnosed atrial fibrillation points to the importance of comprehensive care for such patients and should alert clinicians to detect warning signs of possible early mortality. Clinical Trial Registration: URL: https://www.clinicaltrials.gov . Unique identifier: NCT01090362.

Decentralized residential power generation employing a H2 PEM fuel cell requires that essentially COx free H2 be produced on site by catalytic steam reforming of piped natural gas and then purifying the product H2 (removal of bulk CO2 and dilute CO impurities). Currently, it may be achieved by subjecting the reformed gas to water gas shift reaction followed by (a) removal of all impurities by a PSA process or (b) selective oxidation in a catalytic PROX reactor to reduce only the CO impurity below ~ 10 ppm for use in the fuel cell. The latter approach assumes that the detrimental effect of CO2 on the performance of the fuel cell is minimum. This assumption may not be valid. A recently developed thermal swing sorption enhanced reaction (TSSER) process scheme can be used to combine reformation, shifting, and purification in a compact, single unit operation for this application. The process permits circumvention of the thermodynamic limits of the SMR reaction and permits direct production of fuel cell grade H2 with high recovery and purity, yet operating the SMR reaction at a lower temperature. Simulated performance of the process, preliminary process design for supplying H2 to a 250 KW fuel cell, and first pass costs are described.

Diffuse large B-cell lymphoma (DLBCL) is the most common non-Hodgkin lymphoma (NHL). B-cell NHLs rely on Bruton’s tyrosine kinase (BTK) mediated B-cell receptor signaling for survival and disease progression. However, they are often resistant to BTK inhibitors or soon acquire resistance after drug exposure resulting in the drug-tolerant form. The drug-tolerant clones proliferate faster, have increased metabolic activity, and shift to oxidative phosphorylation, however, how this metabolic programming occurs in the drug-resistant tumor is poorly understood. In this study, we explored for the first time the metabolic regulators of ibrutinib-resistant activated B-cell (ABC) DLBCL using a multi-omics analysis that integrated metabolomics (using high-resolution mass spectrometry) and transcriptomic (gene expression analysis). Overlay of the unbiased statistical analyses, genetic perturbation, and pharmaceutical inhibition was further used to identify the key players contributing to the metabolic reprogramming of the drug-resistant clone. Gene-metabolite integration revealed interleukin four induced 1 (IL4I1) at the crosstalk of two significantly altered metabolic pathways involved in producing various amino acids. We showed for the first time that drug-resistant clones undergo metabolic reprogramming towards oxidative phosphorylation and are modulated via the BTK-PI3K-AKT-IL4I1 axis. Our report shows how these cells become dependent on PI3K/AKT signaling for survival after acquiring ibrutinib resistance and shift to sustained oxidative phosphorylation, additionally, we outline the compensatory pathway that might regulate this metabolic reprogramming in the drug-resistant cells. These findings from our unbiased analyses highlight the role of metabolic reprogramming during drug resistance development. Our work demonstrates that a multi-omics approach can be a robust and impartial strategy to uncover genes and pathways that drive metabolic deregulation in cancer cells.

The rice root system, which primarily consists of adventitious/crown roots (AR/CR) developed from the coleoptile base, is an excellent model system for studying shoot-to-root trans-differentiation process. We reveal global changes in protein and metabolite abundance, and protein phosphorylation in response to an auxin stimulus during CR development. Global proteome and metabolome analyses of developing crown root primordia (CRP) and emerged CRs uncovered that the biological processes associated with chromatin conformational change, gene expression, and cell cycle were translationally regulated by auxin signaling. Spatial gene expression pattern analysis of differentially abundant proteins disclosed their stage-specific dynamic expression pattern during CRP development. Further, our tempo-spatial gene expression and functional analyses revealed that auxin creates a regulatory feedback module during CRP development and activates ethylene biosynthesis exclusively during CRP initiation. Ethylene signaling promotes CR formation by repressing the cytokinin response regulator, OsRR2. Additionally, the (phospho)proteome analysis identified differential phosphorylation of the Cyclin-dependent kinase G-2 (OsCDKG;2), and cell wall proteins, in response to auxin signaling, suggesting that auxin-dependent phosphorylation may be required for cell cycle activation, and cell wall synthesis during root organogenesis. Thus, our study provides evidence for the translational and post-translational regulation during CRP trans-differentiation downstream of the auxin signaling pathway.HighlightGlobal (phospho)proteome and metabolic profiling of rice CRP and CRs uncover differential proteins and metabolites associated with gene expression, cell cycle, ethylene signaling and cell wall synthesis during CR development.

Metastasis, the main cause of breast cancer-associated fatalities, relies on many regular pathways involved in normal cell physiology and metabolism, thus, making it challenging to identify disease-specific therapeutic target(s). Chemically synthesized anti-metastatic agents are preferred for their fast and robust actions. However, these agents have adverse side effects, thus, increasingly favouring the identification of phytocompounds as suitable alternatives. Resveratrol and pterostilbene have long been established as potent anti-cancer agents. Earlier studies from our laboratory documented the anti-cancer activities associated with pterostilbene-isothiocyanate (PTER-ITC), a derivative of pterostilbene. The current study focuses on evaluating the anti-metastatic property of PTER-ITC and the underlying mechanism, by employing in silico, in vitro, and in vivo approaches. The significant anti-metastatic activity of PTER-ITC was observed in vitro against breast cancer metastatic cell line (MDA-MB-231) and in vivo in the 4T1 cell-induced metastatic mice model. Epithelial-mesenchymal transition (EMT), a hallmark of metastasis regulated by the transcription factors, Snail1 and Twist, was found to be reverted in vitro by PTER-ITC treatment. PTER-ITC blocked the activation of NF-κB/p65 and its concomitant nuclear translocation, resulting in the transcriptional repression of its target genes, Snail1 and Twist. PTER-ITC prevented the formation of IKK complex, central to NF-κB activation, by binding to the NEMO-binding domain (NBD) of IKK-β and inhibiting its interaction with NEMO (NF-κB essential modulator). According to our observations, PTER-ITC attenuated NF-κB activation selectively in cancerous cells. In conclusion, this study demonstrated that PTER-ITC is a potent anti-metastatic agent capable of targeting physiologically important pathways in a cancer-specific manner.

BackgroundDiffuse large B-cell lymphoma (DLBCL) is the most common non-Hodgkin lymphoma (NHL). B-cell NHLs rely on Bruton’s tyrosine kinase (BTK) mediated B-cell receptor signaling for survival and disease progression. However, they are often resistant to BTK inhibitors or soon acquire resistance after drug exposure resulting in the drug tolerant form. The drug tolerant clones proliferate faster, have increased metabolic activity, and shift to oxidative phosphorylation; however, how this metabolic programming occurs in the drug resistant tumor is poorly understood.MethodsIn this study, we explored for the first time the metabolic regulators of ibrutinib-resistant activated B-cell (ABC) DLBCL using a ‘multi-omics’ analysis that integrated metabolomics (using high-resolution mass spectrometry) and transcriptomic (gene expression analysis). Overlay of the unbiased statistical analyses, genetic perturbation and pharmaceutical inhibition, were further used to identify the key players that contribute to the metabolic reprograming of the drug resistant clone.ResultsGene-metabolite integration revealed interleukin 4 induced 1 (IL4I1) at the crosstalk of two significantly altered metabolic pathways involved in the production of various amino acids. We showed for the first time that drug resistant clones undergo metabolic reprogramming from glycolysis towards oxidative phosphorylation & is activated via the BTK-PI3K-AKT-IL4I1 axis and can be targeted therapeutically.ConclusionsOur report shows how these cells become dependent on PI3K/AKT signaling for survival after acquiring ibrutinib resistance and shift to sustained Oxidative phosphorylation, additionally we outline the compensatory, pathway that regulates this metabolic reprogramming in the drug resistant cells. These findings from our unbiased analyses highlight the role of metabolic reprogramming during drug resistance development. Furthermore, our work demonstrates that a multi-omics approach can be a powerful and unbiased strategy to uncover genes and pathways that drive metabolic dysregulation in cancer cells.

Sunflower pollen was reported to contain respiratory allergens responsible for occupational allergy and pollinosis. The present study describes the comprehensive characterization of a major sunflower allergen Hel a 6. Natural Hel a 6 was purified from sunflower pollen by anion exchange and gel filtration chromatography. Hel a 6 reacted with IgE-antibodies from 57% of 39 sunflower-sensitized patient sera suggesting it to be a major allergen. The patients were of Indian origin and suffering from pollinosis and allergic rhinitis. Hel a 6 exhibited allergenic activity by stimulating mediator release from basophils. Monomeric Hel a 6 displayed pectate lyase activity. The effect of various physicochemical parameters such as temperature, pH, and calcium ion on the functional activity of Hel a 6 revealed a stable nature of the protein. Hel a 6 was folded, and its melting curve showed reversible denaturation in which it refolded back to its native conformation from a denatured state. Hel a 6 displayed a high degree of sequence conservation with the pectate lyase allergens from related taxonomic families such as Amb a 1 (67%) and Art v 6 (57%). The IgE-cross reactivity was observed between Hel a 6 and its ragweed and mugwort homologs. The cross-reactivity was further substantiated by the mediator release when Hel a 6-sensitized effector cells were cross-stimulated with Art v 6 and Amb a 1. Several putative B cell epitopes were predicted and mapped on these 3 allergens. Two antigenic regions were found to be commonly shared by these 3 allergens, which could be crucial for cross-reactivity. In conclusion, Hel a 6 serves as a candidate molecule for diagnosis and immunotherapy for weed allergy.

In this manuscript, we study the tribological characteristics of a carbon nanotube (CNT) moving on a graphene sheet using molecular dynamics simulations. The CNT is driven by a force acting due to a spring that itself is pulled with a constant velocity. While the interaction between the carbon-carbon atoms of both CNT and graphene are modeled using Tersoff potential, the cross-interaction between the carbon atoms of CNT and those of graphene are modeled through the Lennard-Jones potential. A dynamics based scheme is proposed in this manuscript that helps in determining the different regimes of the moving CNT. Using the proposed scheme, we show that the tribological and dynamical characteristics of the CNT-graphene system depend on the velocity with which the spring is pulled—the stick-roll motion of the CNT at low pulling velocity changes to a combination of rolling-sliding at moderate pulling velocity. Subsequently, one observes sliding motion of CNT at large pulling velocities. Further, in order to elucidate the role of thermal fluctuations we study the CNT-graphene system at very low (1K) and very high temperatures (300K). The results indicate that the tribological characteristics depend on the interaction of the CNT-graphene system with the temperature of the surrounding environment. At high temperature, thermal fluctuations are high and these fluctuations overshadow the normal dynamics of the CNT-graphene system. Our results suggest that it is possible to minimize frictional losses in the CNT-graphene system by suitably altering the driving forces.

Objective There exists a complex interaction between alcohol and stress on brain and behavior. Alcohol and stress are both known to affect memory. Whether stress and alcohol together can modulate memory functions in adolescent rats is not known. In the present study, effects of repeated unpredictable stress (RUPS) on contextual fear conditioning, a hippocampus-related memory function, were investigated in alcohol-treated adolescent rats. Methods Rats were divided into four experimental groups: group i - saline-treated non-stressed rats (sal no stress), group ii - alcohol-treated non-stressed rats (alc no stress), group iii - saline-treated rats subjected to stress (sal + RUPS), group iv - alcohol-treated rats subjected to stress (alc + RUPS). All rats were trained in the fear conditioning paradigm, and 24 h later were tested for contextual fear conditioning in the conditioning chamber, and nonspecific fear memory in a modified chamber. Results Stress, in the presence or absence of alcohol, did not alter nonspecific fear. RUPS exposure did not affect contextual freezing in vehicle-treated adolescent rats. Compared to vehicle-treated non-stressed rats, alcohol-treated non-stressed rats showed significant impairments in contextual freezing. Alcohol-treated RUPS rats performed better in the contextual freezing task than alcohol-treated non-stressed rats. Conclusion RUPS exposure did not alter contextual fear conditioning in adolescent rats. Alcohol significantly reduced contextual fear memory in non-stressed rats. Alcohol-treated RUPS rats showed significantly better memory than alcohol-treated non-stressed rats. Together, these data suggest resiliency to stress-induced memory impairment in adolescent rats, and RUPS exposure causes blunting of alcohol's negative effects on contextual fear conditioning.

The black pepper, most commonly used in Indian cuisines for ages, is considered as “king of spices.” The present study evaluates the anticancer potential of black pepper and its main constituent, i.e. alkaloid piperine, against human leukemia cell line, K-562 cells. Gas chromatography–mass spectrometry (GC–MS) analysis confirmed the presence of piperine in black pepper extract. The methanolic extract of black pepper (BP-M) and pure piperine (PIP) showed a strong cytotoxic effect against this cell line. Both BP-M and PIP generated apoptotic bodies in K-562 cells and caused nuclear condensation as visualized by fluorescent microscopy, which was further confirmed by flow cytometry analysis. BP-M and PIP also generated reactive oxygen species in K-562 cells as established by flow cytometry. The translation of Bax, caspase-3 and caspase-9 genes was found to be upregulated with subsequent downregulation of Bcl-2 gene. The anti-proliferative effect of both BP-M and PIP was also observed by trypan blue staining and was further confirmed by the downregulated expression of proliferating cell nuclear antigen (PCNA). The molecular docking studies showed the binding of PIP with PCNA and Bcl-2 and supported the in vitro findings. The docking studies also proposed the binding of PIP to ADP binding pocket of Apaf-1 protein. Taken together, these findings signify the anticancer potential of both black pepper and PIP, thus proposing black pepper as a potent nutraceutical for preventing the progression of chronic myeloid leukemia.

Chronic activation of B-cell receptor (BCR) signaling via Bruton tyrosine kinase (BTK) is largely considered to be one of the primary mechanisms driving disease progression in B–Cell lymphomas. Although the BTK-targeting agent ibrutinib has shown promising clinical responses, the presence of primary or acquired resistance is common and often leads to dismal clinical outcomes. Resistance to ibrutinib therapy can be mediated through genetic mutations, up-regulation of alternative survival pathways, or other unknown factors that are not targeted by ibrutinib therapy. Understanding the key determinants, including tumor heterogeneity and rewiring of the molecular networks during disease progression and therapy, will assist exploration of alternative therapeutic strategies. Towards the goal of overcoming ibrutinib resistance, multiple alternative therapeutic agents, including second- and third-generation BTK inhibitors and immunomodulatory drugs, have been discovered and tested in both pre-clinical and clinical settings. Although these agents have shown high response rates alone or in combination with ibrutinib in ibrutinib-treated relapsed/refractory(R/R) lymphoma patients, overall clinical outcomes have not been satisfactory due to drug-associated toxicities and incomplete remission. In this review, we discuss the mechanisms of ibrutinib resistance development in B-cell lymphoma including complexities associated with genomic alterations, non-genetic acquired resistance, cancer stem cells, and the tumor microenvironment. Furthermore, we focus our discussion on more comprehensive views of recent developments in therapeutic strategies to overcome ibrutinib resistance, including novel BTK inhibitors, clinical therapeutic agents, proteolysis-targeting chimeras and immunotherapy regimens.

Aims: To assess the antioxidant and antibacterial activities of two locally available natural honey samples, and determine the total phenolic content (TPC) of the honeys.Methods: Antibacterial activity of two local natural honey samples (citrus honey: LNH1 and mango honey: LNH2) was determined against the clinical bacterial isolates of Staphylococcus aureus, Pseudomonas aeruginosa, Salmonella enterica serovar Typhi, and Escherichia coli ATCC 25922 strain, by agar-well diffusion method. The antioxidative activity of the honey samples (LNH1 and LNH2) was determined in DPPH system, and the IC50 values were calculated. The bioactive components present in the test honey samples were assessed qualitatively, and the TPC was quantified.Results: The honey samples showed the presence of different bioactive components (flavonoids, steroids, phenol, terpenoids and quinone); the TPCs were estimated in LNH1 and LNH2 were 195.5 μg/ml and 333 μg/ml, respectively. In DPPH system, the scavenging activities of LNH1 and LNH2 ranged 43.46–75.18% and 36.09–77. 23%, respectively, with respective IC50 values of 3.83 mg/ml and 6.75 mg/ml. The test honey samples showed antibacterial activity both against gram-positive (Staph. aureus) and gram-negative (Ps. aeruginosa and S. enterica serovar Typhi) bacterial pathogens, and against the E. coli ATCC 25922 strain.Conclusion: The locally available natural honey may be used as rich source of antioxidant and can be used as effective antibacterial agent in order to combat the bacterial infection to humans.

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) led to coronavirus disease 2019 (COVID-19) pandemic affecting nearly 71.2 million humans in more than 191 countries, with more than 1.6 million mortalities as of 12 December, 2020. The spike glycoprotein (S-protein), anchored onto the virus envelope, is the trimer of S-protein comprised of S1 and S2 domains which interacts with host cell receptors and facilitates virus-cell membrane fusion. The S1 domain comprises of a receptor binding domain (RBD) possessing an N-terminal domain and two subdomains (SD1 and SD2). Certain regions of S-protein of SARS-CoV-2 such as S2 domain and fragment of the RBD remain conserved despite the high selection pressure. These conserved regions of the S-protein are extrapolated as the potential target for developing molecular diagnostic techniques. Further, the S-protein acts as an antigenic target for different serological assay platforms for the diagnosis of COVID-19. Virus-specific IgM and IgG antibodies can be used to detect viral proteins in ELISA and lateral flow immunoassays. The S-protein of SARS-CoV-2 has very high sequence similarity to SARS-CoV-1, and the monoclonal antibodies (mAbs) against SARS-CoV-1 cross-react with S-protein of SARS-CoV-2 and neutralize its activity. Furthermore, in vitro studies have demonstrated that polyclonal antibodies targeted against the RBD of S-protein of SARS-CoV-1 can neutralize SARS-CoV-2 thus inhibiting its infectivity in permissive cell lines. Research on coronaviral S-proteins paves the way for the development of vaccines that may prevent SARS-CoV-2 infection and alleviate the current global coronavirus pandemic. However, specific neutralizing mAbs against SARS-CoV-2 are in clinical development. Therefore, neutralizing antibodies targeting SARS-CoV-2 S-protein are promising specific antiviral therapeutics for pre-and post-exposure prophylaxis and treatment of SARS-CoV-2 infection. We hereby review the approaches taken by researchers across the world to use spike gene and S-glycoprotein for the development of effective diagnostics, vaccines and therapeutics against SARA-CoV-2 infection the COVID-19 pandemic.

Background Chimaphila umbellata (CU) is a small perennial plant whose roots and leaves are used in the preparation of homeopathic medicines. In this study, the anti-proliferative and apoptotic effects of CU extract, obtained from its homeopathic mother tincture, were evaluated in human breast cancer cell lines. Methods MTT assay was used to evaluate the dose- and time-dependent cytotoxicity of the extract in MCF-7 and HEK293 cells. The anti-proliferative effect of the extract was evaluated using the clonogenic and wound healing assays. The anti-angiogenic activity of the drug was evaluated using the chick chorioallantoic membrane assay. The mode of cell death was analyzed using Annexin V and PI staining assays through flow cytometry. In addition, the expression patterns of associated genes were evaluated using immunoblot analysis. Results CU extracts exerted anti-proliferative effect in breast cancer cell line, i.e., MCF-7 cells, by inhibiting their growth and migration. The extract also demonstrated significant anti-angiogenic ability, limiting the de-novo blood vessel growth and development in chick embryos. The extract was found to increase caspase-independent necroptosis by involving RIP1/RIP3 kinases and MLKL proteins. This finding was further confirmed by using NEC-1 (an inhibitor of necroptosis), which significantly abolished the CU extract-induced necroptotic effect. Conclusions The CU extract exhibits great potential in preventing breast cancer and thus warrants further investigations in vivo.

Cell cultures of Asian pear (Pyrus pyrifolia) are known to produce benzoate-derived biphenyl phytoalexins upon elicitor treatment. Although the downstream pathway for biphenyl phytoalexin biosynthesis is almost known, the upstream route of benzoic acid biosynthesis in pear has not been completely elucidated. In the present work, we report benzaldehyde synthase (BS) activity from yeast extract-treated cell suspension cultures of P. pyrifolia. BS catalyzes the in vitro conversion of trans-cinnamic acid to benzaldehyde using a non-oxidative C2 -side chain cleavage mechanism. The enzyme activity was strictly dependent on the presence of a reducing agent, dithiothreitol being preferred. C2 -side chain shortening of the cinnamic acid backbone resembled the mechanisms catalyzed by 4-hydroxybenzaldehyde synthase (HBS) activity in Vanilla planifolia and salicylaldehyde synthase (SAS) activity in tobacco and apple cell cultures. A basal BS activity was also observed in the non-elicited cell cultures. Upon yeast extract-treatment, a 13-fold increase in BS activity was observed when compared to the non-treated control cells. Moreover, feeding of the cell cultures with trans-cinnamic acid, the substrate for BS, resulted in an enhanced level of noraucuparin, a biphenyl phytoalexin. Comparable accumulation of noraucuparin was observed upon feeding of benzaldehyde, the BS product. The preferred substrate for BS was found to be trans-cinnamic acid, for which the apparent Km and Vmax values were 0.5 mM and 50.7 pkat mg-1 protein, respectively. Our observations indicate the contribution of BS to benzoic acid biosynthesis in Asian pear via the CoA-independent and non-β-oxidative route.

This study investigates the production of biodiesel from activated sludge of milk processing plant as a low cost feedstock. The method involves lipid extraction, refining and optimization of biodiesel from activated sludge under laboratory conditions. These results were scaled up to a pilot plant was analyzed. In the laboratory study, the sludge yielded 18.81 wt% of crude lipid. It is degummed and dewaxed to get refined oil that is used for the production of biodiesel. The optimal reaction parameters were 0.8 wt% catalyst; a temperature of 55 °C; 6:1 methanol to oil molar ratio for 40 min. The process is scaled up to a prototype demonstration plant with a batch size of 10 kg. The dried sludge yielded 16.2 ± 1.3 wt% and 13.64 ± 0.8 wt% of crude lipid and biodiesel respectively. Biodiesel made in pilot plant is compared with ASTM standards. Keywords: Activated sludge, Extraction, Biodiesel, Scale up

The present work investigates the production of biodiesel from Euglena sanguinea microalgal bio-oil using calcium methoxide as a heterogeneous catalyst. The catalyst was synthesized and cha...

This manuscript describes the cloning and functional characterization of a biphenyl phytoalexin biosynthetic gene, 3,5-dihydroxybiphenyl O-methyltransferase from elicitor-treated cell cultures of scab resistant apple cultivar 'Florina'. Apples belong to the subtribe Malinae of the Rosaceae family. Biphenyls and dibenzofurans are the specialized phytoalexins of Malinae, of which aucuparin is the most widely distributed biphenyl. The precursor of aucuparin, 3,5-dihydroxybiphenyl, is a benzoate-derived polyketide, which is formed by the sequential condensation of three molecules of malonyl-CoA and one molecule of benzoyl-CoA in a reaction catalyzed by biphenyl synthase (BIS). This 3,5-dihydroxybiphenyl then undergoes sequential 5-O-methylation, 4-hydroxylation, and finally 3-O-methylation to form aucuparin. A cDNA encoding O-methyltransferase (OMT) was isolated and functionally characterized from the cell cultures of scab-resistant apple cultivar 'Florina' (Malus domestica cultivar 'Florina'; MdOMT) after treatment with elicitor prepared from the apple scab causing fungus Venturia inaequalis. MdOMT catalyzed the regiospecific O-methylation of 3,5-dihydroxybiphenyl at the 5-position to form 3-hydroxy-5-methoxybiphenyl. The enzyme showed absolute substrate preference for 3,5-dihydroxybiphenyl. The elicitor-treated apple cell cultures showed transient increases in the MdOMT (GenBank ID MF740747) and MdBIS3 (GenBank ID JQ390523) transcript levels followed by the accumulation of biphenyls (aucuparin and noraucuparin) and dibenzofuran (eriobofuran) phytoalexins. MdOMT fused with N- and C-terminal yellow fluorescent protein showed cytoplasmic localization in the epidermis of Nicotiana benthamiana leaves. In scab inoculated greenhouse-grown 'Florina' plants, the expression of MdOMT was transiently induced in the stem followed by the accumulation of biphenyl phytoalexins.

This research article investigates the extraction of bio-oil from Chlorella vulgaris algae which is then subjected to biodiesel production. To evaluate the maximum oil content, four different pretr...

The knowledge of water temperature produced from a geothermal reservoir and its composition is of utmost importance in designing utilization strategies, the surface production facilities and in selecting the material to be used. Unai hot springs are located in the southern part of Gujarat, India with discharge temperatures varying from 51 to 56 °C. With the aim of developing Unai as a potential geothermal field and exploiting it in the future, geochemical and geothermometrical study was undertaken. The samples were collected from various Unai geothermal location and analysis of chemical composition of water obtained from different wells was done. The concentration of Silica, carbonate and ions like Na and Cl have been analyzed to delineate the path of water movement in the subsurface and classify the reservoir based on the enthalpy. The ratio of the concentration of ions like Na+ and K+ and the relative proportions of various sets of ions were also used to characterize the geothermal reservoir and the reservoir fluid by the use of ternary diagrams. Na–K–Ca, Chalcedony, quartz, and Silica geo-thermometers have been studied. The present study also envisages the importance of graphical representations like Piper diagram, Scholler etc. to determine variation in hydrochemical facies and to understand the evolution of hydrochemical processes in the Unai geothermal field respectively. The study yields the conclusion that the reservoir under consideration is a low enthalpy reservoir with temperature ranging from 60 to 80 °C.

Kaempferol, a natural flavonoid, has the beneficial effects of preserving pancreatic β-cell mass and function, but its action on β-cell lipid metabolism still remains elusive. Recently, autophagy has been reported to play a major role in lipid metabolism in various cell types, but its role in pancreatic β-cell's lipid metabolism is rarely reported. Here, we investigated the role of kaempferol-induced autophagy in inhibition of lipid stores, ER stress and β-cell dysfunction in palmitic acid-challenged RIN-5F cells and isolated pancreatic islets. The lipid-lowering effect of kaempferol was determined by Oil Red O staining, triglyceride assay, BODIPY labeling, RT-PCR and immunoblot analysis of PLIN2 (the lipid droplet coat protein) expression. Further, the involvement of AMPK/mTOR-mediated lipophagy was established by pharmacological and genetic inhibitors of autophagy and AMPK. The co-localization studies of lipid droplets with autophagosomes/lysosomes by BODIPY-MDC-LysoTracker co-staining, LC3/BODIPY labeling and LC3/PLIN2 double immunolabeling further strengthened the findings. Kaempferol treatment exhibited decreased lipid stores and increased co-localization of lipid droplets with autophagosomes and lysosomes in palmitic acid-challenged β-cells. Moreover, inhibition of autophagy led to decreased co-localization and increased lipid droplets accumulation. Kaempferol-induced alleviation of ER stress and β-cell dysfunctions was established by immunoblot analysis of CHOP-10 (a key mediator of cell death in response to ER stress) and insulin content/secretion analysis respectively. Together, these findings suggest that kaempferol prevents ectopic lipid accumulation and ER stress, thus restoring β-cell function through AMPK-mediated lipophagy. The current data implies that kaempferol may be a potential therapeutic candidate to prevent obesity-linked diabetic complications.

Shale Gas exploration and production plays an essential role in ensuring energy security and economic stability. Tapping these reserves using cost-effective technologies can really change the India’s energy supply and value chain. Cambay Shale of Cambay Basin has been proved as the most prominent Indian Shale reserve and in this work an attempt has been made to understand the geographical and mineralogical distribution of this Shale. A sample study on Cambay Shale of North Tectonic Block of Cambay Basin has been ascertained to understand the mineralogy, porosity and maturity with respect to depth. The study involves the use of integrated measurement techniques mainly high-pressure porosity (HPP), scanning electron microscopy (SEM) and Fourier transform electron microscopy (FTIR). Approximately, 13 samples were analysed to understand the mineralogy of this shale and it was observed that it is very clayey in nature with major traces of Illite and minor traces of rutile and kaolinite. The other measurements (i.e., SEM and HPP) were done on two specific samples which confirm the presence of clay-rich porous minerals with high organic matter content.

We report the formation of crack patterns in drying films of Laponite-NaCl solution. Crack patterns that develop upon drying aqueous Laponite-NaCl solution change drastically as the amount of NaCl is varied in the solution. In this work, we have investigated the effect of NaCl on drying films of aqueous solution of Laponite under two conditions: (i) when the film is bounded by a wall, as in Petri dish experiments and (ii) when the film does not have any boundary, as in experiments with droplets. In order to obtain insights into the effect of the substrate, the experiments have been done with two different substrates of different hydrophobicities, polypropylene and glass. The formation of crack patterns has been explained on the basis of the wetting and spreading properties of the solution on these substrates and the effect of salt on colloidal aggregation. In this work, we have shown that the presence of salt in aqueous Laponite solution can induce crack patterns depending on the nature of the substrate. Another important aspect of this work is the role of NaCl in crack inhibition in desiccating films of aqueous Laponite, in the presence of static electric field. This effect can be utilized to suppress undesirable crack formation in many applications.

The search for a suitable low cost feedstock for the production of biodiesel has resulted in biodiesel being produced from bio-solids (scum) taken from the tannery effluent treatment facility. The effectively extracted oil was subjected to combined esterification and transesterification using an acid catalyst. The process was optimized for parameters like catalyst concentration, temperature, time, oil to methanol molar ratio and stirring rate. The result was the yield of 0.55 kg of biodiesel from 1 kg wet scum. The physicochemical properties of the produced biodiesel are in the acceptable range of fuel used in diesel engines. The paper also addresses the quality issues regarding minor components like heavy metals and aromatic contents.

In this paper, a three-dimensional model for hydrogen absorption in a metal alloy has been developed, validated against the experimental data in the literature, and then applied to a novel design for a hydrogen storage unit. The proposed design is similar to the fuel cell stack, but here the Membrane Electrode Assembly (MEA) has been replaced by a metal hydride (MH) reactor placed between the flow-field plates. These are stacked together to achieve the required amount of hydrogen storage. The flow-field plates have channels engraved on one side for hydrogen supply and on the other, for coolant/heating medium. It is known that the effectiveness of a hydrogen storage unit is directly related to its heat transfer area, and therefore, the choice of its geometry is very important. The larger the size, the more the resistance to heat transfer. Although, the internal tubular heat exchangers have proven to be effective in heat transfer, they pose severe challenges such as cooling/heating medium leakage due to tube erosion, stresses generated, etc. and they displace the active metal hydride from the tank. The present stacked MH reactor configuration helps to overcome these challenges by stacking small MH reactors together and there is no chance of the cooling/heating medium leaking into the metal hydride. Numerical simulations were performed to investigate the effect of coolant flow rate and percentage of flow-field plate rib area exposed to the MH reactor on temperature evolution and the amount of hydrogen stored. Further, a detailed study was carried out to understand the effect of compartmentalization of the MH reactor on temperature distribution. The results revealed that compartmentalization substantially helps to uniformly distribute the temperature in the metal bed, which is very important to maintain uniform utilization of the metal powder. Consequently, the uniform metal powder density for repeated absorption-desorption cycles without significant loss of its hydrogen storage capabilities.