Chemical composition, antimicrobial activity and cytotoxicity of Murraya paniculata (L.) Jack leaf essential oil from Assam, India: the effect of oil on cellular morphology of micro-organisms.

Murraya paniculata (L.) Jack is commonly cultivated as ornamental plant in Assam and has been used as spice and phytomedicine traditionally for many healthcare purposes. The therapeutic potential and chemical constituents of the essential oil of M. paniculata leaf was investigated against several pathogenic microbial species and human cancer cell lines. 29 chemical compounds were identified by GC-MS analysis from the essential oil representing 97.62% of the oil. The major compound identified was caryophyllene (20.93%). Leaf essential oil exhibited promising antibacterial activity against Mycobacterium smegmatis (MIC = 4 µg/mL) and Pseudomonas aeruginosa (MIC = 4 µg/mL). Best anticancer activity of the oil was observed for HeLa cells (IC50 = 6.28 µg/mL). Further, scanning electron microscopic studies revealed that the oil kills micro-organisms with the deformation of cellular morphology on treatment of the oil. Thus, the essential oil of M. paniculata leaf can be an excellent alternative for development of new antimicrobials and anticancer chemotherapeutic agents for the pharmaceutical industries.

Integrated Genomic and Functional Characterization of the Anti-diabetic Potential of Arthrobacter sp. SW1.

For decades, bacterial natural products have served as valuable resources for developing novel drugs to treat several human diseases. Recent advancements in the integrative approach of using genomic and functional tools have proved beneficial in obtaining a comprehensive understanding of these biomolecules. This study presents an in-depth characterization of the anti-diabetic activity exhibited by a bacterial isolate SW1, isolated from an effluent treatment plant. As a primary screening, we assessed the isolate for its potential to inhibit alpha-amylase and alpha-glucosidase enzymes. Upon confirmation, we further utilized LC-MS, ESI-MS/MS, and NMR spectroscopy to identify and characterize the biomolecule. These efforts were coupled with the genomic assessment of the biosynthetic gene cluster involved in the anti-diabetic compound production. Our investigation discovered that the isolate SW1 inhibited both α-amylase and α-glucosidase activity. The chemical analysis suggested the production of acarbose, an anti-diabetic biomolecule, which was further confirmed by the presence of biosynthetic gene cluster "acb" in the genome. Our in-depth chemical characterization and genome mining approach revealed the potential of bacteria from an unconventional niche, an effluent treatment plant. To the best of our knowledge, it is one of the first few reports of acarbose production from the genus Arthrobacter.

Antiviral compound screening, peptide designing, and protein network construction of influenza a virus (strain a/Puerto Rico/8/1934 H1N1).

Plant-based antiviral therapy is the current need for holistic health care management, which can be achieved through screening of phytochemicals and designing of antiviral peptides. There exist certain host's factors which are directly involved for rapid viral replication causing worldwide pandemic. A total of 177 phytochemicals from Ocimum sanctum (L.), Tinospora cordifolia (Thunb.) Miers, Cinnamomum camphora (L.) J. Presl., Allium sativum (L.), Curcuma longa (L.), and Aloe vera (L.) Burm. f. were evaluated for their affinity to all viral proteins of H1N1. Applying drug filters and keeping the threshold of such filters relative to the standards, 82 compounds were found suitable for further analysis. Consensus scoring system was used for screening top ligands from 82 compounds, which screened the top 12 compounds. Highly conserved regions (>80%) which were hydrophilic, flexible, antigenic, and also charged were screened out as potent antiviral peptides. The viral proteins were taken as the targets for the modeled peptides for protein-protein docking. Further, host-pathogen interacting network was constructed to unveil host factors involved in viral replication, from which unique protein clusters representing their involvement in viral reproduction were selected through mapping with pathway databases. Twelve compounds and five peptides were found to be highly effective against all the proteins of H1N1. Based on the uniqueness, 13 clusters of proteins were obtained which are engaged in cellular process, namely, viral reproduction, fructose-6-phosphate metabolism, nitrogen compound metabolism, biosynthesis, cellular process, oligodendrocyte development, localization, multiorganism process, primary metabolism, response to unfolded protein, metabolism, and response to protein and catabolism.

Multicolored Protein Nanoparticles: Synthesis, Characterization, and Cell Uptake.

Synthesis, characterization, and applications of strongly fluorescent, multicolored protein nanoparticles (GlowDots) are reported here. Bovine serum albumin was cross-linked under controlled conditions to form nanoparticles, where particle size was controlled from 20 to 100 ± 10 nm by choosing appropriate reaction conditions. The absorption as well as the emission wavelengths were controlled without changing the particle size, unlike quantum dots. Each GlowDot was loaded with up to 214 ± 50 chromophores, and hence, the particles have high molar absorptivities (106 M-1 cm-1) as well as high brightness (105 to 106 M-1 cm-1). A large number of functional groups cover the particle surface and these are further functionalized to enhance cellular uptake. GlowDots that were labeled with fluorescein and functionalized with taurine, for example, were quickly taken up by HeLa, MDA-MB-231, PC3, and L6 myoblast cells, as interrogated by fluorescence imaging studies. GlowDots were biocompatible, size tunable, biodegradable, strongly fluorescent, and stable for months at room temperature, and they may serve as substitutes for quantum dots in a variety of practical applications.

Diastereoselective synthesis of tetrahydropyrans via Prins-Ritter and Prins-arylthiolation cyclization reactions.

An efficient method has been developed for the synthesis of two new classes of tetrahydropyran derivatives comprising amide, tetrazole or benzothiazole moieties via a three-component reaction of 6-methylhept-5-en-2-ol, arylaldehydes and nitriles/thiols in the presence of a tetrafluoroboric acid diethyl ether complex. The reaction proceeds via the formation of an oxocarbenium ion. This protocol is highly diastereoselective and only single diastereomer has been isolated in each case.

Evaluation of commercial importance of endophytes isolated from Argemone mexicana and Papaver rhoeas.

The paper industry is a composite one constituting different types of mills, processes, and products. The paper industries consume large amounts of resources, like wood and water. These industries also create huge amounts of waste that have to be treated. In our study, 23 endophytic bacteria were isolated from Argemone mexicana, and 16 endophytic bacteria were isolated from Papaver rhoeas. Seventeen and 15 bacterial endophytes from A. mexicana and P. rhoeas, respectively, showed cellulose-degrading activity. The biochemical and molecular characterization were done for endophytic bacteria with cellulolytic activity. The consortium of cellulose-degrading endophytic bacteria from A. mexicana showed endoglucanase activity (0.462 IU/ml) and FPCase enzyme activity (0.269 IU/ml) and from P. rhoeas gave endoglucanase activity (0.439 IU/ml) and FPCase enzyme activity (0.253 IU/ml). Degraded carboxy methylcellulose and filter paper were further treated by Saccharomyces cerevisiae and bioethanol was produced. Cellulose-degrading endophytic bacteria were also tested for auxin, siderophore production, and phosphate solubilization activities. Individual cellulose-degrading endophytic bacteria with plant growth-promoting activities were used as biofertilizers, tested for plant growth-promoting activities using Basmati Pusa 1121 rice, and plant growth parameters were recorded. The degraded paper enhances the growth of rice plants. Selected bacterial endophytes and their consortia from A. mexicana and P. rhoeas were powerful cellulose degraders, which can be further employed for ethanol production and as significant biofertilizers in agriculture.

Efficient Ru-Catalyzed Electrochemical Homo- and Heterocoupling Reaction of Terminal Alkynes: Synthesis, In Vitro Anticancer Activity, and Docking Study.

With the objective to identify novel anticancer leads, herein ruthenium-catalyzed electrochemical homo- and heterocoupling reactions of terminal alkynes have been developed for the synthesis of the desired products. Among the synthesized 1,3-diynes, some of them were rigorously examined for possible in vitro anticancer activity against HeLa (human cervical cancer) and L6 normal (rat skeletal muscle) cell lines. Additionally, the docking study was also performed toward 16 ovarian cancer targets with binding affinity calculations with respect to the standard. To the best of our knowledge, this is the first scientific report on the ruthenium-catalyzed electrochemical homocoupling reaction between terminal alkynes with its in vitro anticancer and in silico docking studies.

Major chemical constituents from Illicium griffithii Hook. f. & Thoms of North East India and their cytotoxicity and antimicrobial activities.

Illicium griffithii Hook. f. & Thoms is an endemic medicinal plant of North East India found in the Eastern Himalayan region of biodiversity mega centre. Herein, chemical investigation of I. griffithii, afforded five compounds and their structures were determined through extensive use of NMR, HRMS, and FT-IR spectroscopy. The complete proton-proton, proton-carbon coupling network of compound 1 was determined using 1H-1H COSY, HSQC and NOESY NMR experiments. All the compounds were evaluated for their cytotoxic activity by MTT assay and antimicrobial activity by Agar well diffusion method. Compound 1 exhibited significant cytotoxicity activity against Lung cancer (A549) and pancreatic cancer (MIAPaCa2) cell lines with IC50 values of 15.01 ± 2.69 µg/mL and 47.77 ± 2.38 µg/mL, respectively. Further, the compound 1 exhibited good antimicrobial activities against Escherichia coli and Candida albicans with MIC 7.50 ± 0.28 µg/mL and 7.50 ± 0.86 µg/mL, respectively. The other isolated compounds along with the extracts of I. griffithii also displayed moderate anticancer and antimicrobial activities against respective strains. To the best of our knowledge, this is the first study of isolation of compounds from bark, wood, and leaf along with cytotoxicity and antimicrobial activities of I. griffithii from the North Eastern region of India and could be a potential herbal medicine in near future.

Anticancer and antimicrobial compounds from Croton caudatus Gieseler and Eurya acuminata DC: Two edible plants used in the traditional medicine of the Kuki tribes.

Eurya acuminata DC and Croton caudatus Gieseler are two ethno-medicinal plants used by Kuki community of North East India. From these plants, we have characterized fifteen phytochemicals (1-15) by extensive use of chromatographic and spectroscopic methods. They were also tested for in vitro cytotoxic effects against A549 and MIAPACA2 cell lines and antimicrobial activities against Mycobacterium smegmatis and Candida albicans. All compounds showed moderate activity against the MIAPACA2 cell lines. Compounds tricosan-1-ol (6), octacosanoic acid (7), ß-sitosterol (10) and (E)-dodec-3-en-1-ol (14) exhibited promising activity against A549 cell lines with IC50 of 16.72, 4.5, 4.42 and 4.5 µg/ml respectively. Further, hexatriacontan-1-ol (2) exhibited lowest MIC of 50 µg/ml against C. Albicans and henicosan-1-ol (3) at 25 µg/ml against M. smegmatis. They were also screened through docking analysis against two Phosphatidylinositol-3-Kinase nodal proteins and three feedback loop proteins of cancers. Thus, this study validates their traditional uses as herbal anticancer and antimicrobial agents.

Bio-transformation of artemisinin using soil microbe: Direct C-acetoxylation of artemisinin at C-9 by Penicillium simplissimum.

Potent antimalarial compound artemisinin, 1 was bio-transformed to C-9 acetoxy artemisinin, 2 using soil microbe Penicillium simplissimum along with C-9 hydroxy derivative 3. The products were characterized using high field NMR and MS-MS data. The absolute stereochemistry of the newly generated chiral centers has been ascertained by COSY and 1D NOESY experiments. This is the first Letter of direct C-acetoxylation of artemisinin using microbial strains.

Au/NiFe2O4 nanoparticle-decorated graphene oxide nanosheets for electrochemical immunosensing of amyloid beta peptide.

In the present work, an electrochemical immunosensor has been fabricated for the detection of amyloid beta peptide (ßA1--42) based on a gold nanoparticle/nickel ferrite decorated graphene oxide-chitosan nanocomposite (Au/NiFe2O4@GO-Ch) modified glassy carbon electrode (GCE) as an effective sensing platform. ßA1-42 has been analyzed as a potential biomarker for its application in Alzheimer's disease monitoring. The combination of highly conducting Au and NiFe2O4 nanoparticles on two-dimensional GO nanosheets provides an excellent platform for sensitive and selective sensing applications. A miniaturized Au/NiFe2O4@GO-Ch/GCE immunosensor was prepared by immobilization of ßA antibody onto Au//NiFe2O4@GO-Ch/GCE via carbodiimide coupling. Various characterization techniques were utilized in the study to estimate the morphological and electronic attributes of the components used to fabricate the immunosensor. Differential pulse voltammetry (DPV) was performed to study the amperometric response of the developed immunosensor as a function of ßA1-42 concentration. The DPV results confirmed that the immunosensor detected ßA1-42 selectively and demonstrated a wide linear range from 1 pg mL-1 to 1 ng mL-1 and a detection limit of 3.0 pg mL-1. Furthermore, the immunosensor also indicated its clinical viability by detecting ßA1-42 in cerebrospinal fluid.

Acid mine drainage in an Indian high-sulfur coal mining area: Cytotoxicity assay and remediation study.

Opencast mining causes significant environmental concern due to acid mine drainage (AMD) caused by the oxidation of pyrites and other sulfur-bearing minerals. The present study intends to determine the seasonal variability of AMD in the affected area of the Ledo opencast mining, the cytotoxicity of the AMD, and the AMD remediation process. The physicochemical properties of the collected samples were analyzed by using laboratory-based methods and sophisticated instrumental tools. The cytotoxicity study of AMD water was performed by using different cell lines such as normal rat muscle and human carcinoma cells. The study demonstrates that the mine water samples have high conductivity (1.30-2.49 ms cm-1) with high total dissolved solids (1068-1339 ppm) which can change the ionic composition of water. The concentration level of trace elements are also found to be higher than the permissible limit during monsoon season. A simple laboratory-based remediation process of AMD has been carried out in the current study by using size segregated pulverized limestone and the process reveals the decrease in elemental concentrations of AMD water. This study will be useful to develop a remediation technique to minimize the concentration levels of hazardous elements and ions in the AMD water.

Molecular Docking: Challenges, Advances and its Use in Drug Discovery Perspective.

Molecular docking is a process through which small molecules are docked into the macromolecular structures for scoring its complementary values at the binding sites. It is a vibrant research area with dynamic utility in structure-based drug-designing, lead optimization, biochemical pathway and for drug designing being the most attractive tools. Two pillars for a successful docking experiment are correct pose and affinity prediction. Each program has its own advantages and drawbacks with respect to their docking accuracy, ranking accuracy and time consumption so a general conclusion cannot be drawn. Moreover, users don't always consider sufficient diversity in their test sets which results in certain programs to outperform others. In this review, the prime focus has been laid on the challenges of docking and troubleshooters in existing programs, underlying algorithmic background of docking, preferences regarding the use of docking programs for best results illustrated with examples, comparison of performance for existing tools and algorithms, state of art in docking, recent trends of diseases and current drug industries, evidence from clinical trials and post-marketing surveillance are discussed. These aspects of the molecular drug designing paradigm are quite controversial and challenging and this review would be an asset to the bioinformatics and drug designing communities.

Established and In-trial GPCR Families in Clinical Trials: A Review for Target Selection.

The largest family of drug targets in clinical trials constitute of GPCRs (G-protein coupled receptors) which accounts for about 34% of FDA (Food and Drug Administration) approved drugs acting on 108 unique GPCRs. Factors such as readily identifiable conserved motif in structures, 127 orphan GPCRs despite various de-orphaning techniques, directed functional antibodies for validation as drug targets, etc. has widened their therapeutic windows. The availability of 44 crystal structures of unique receptors, unexplored non-olfactory GPCRs (encoded by 50% of the human genome) and 205 ligand receptor complexes now present a strong foundation for structure-based drug discovery and design. The growing impact of polypharmacology for complex diseases like schizophrenia, cancer etc. warrants the need for novel targets and considering the undiscriminating and selectivity of GPCRs, they can fulfill this purpose. Again, natural genetic variations within the human genome sometimes delude the therapeutic expectations of some drugs, resulting in medication response differences and ADRs (adverse drug reactions). Around ~30 billion US dollars are dumped annually for poor accounting of ADRs in the US alone. To curb such undesirable reactions, the knowledge of established and currently in clinical trials GPCRs families can offer huge understanding towards the drug designing prospects including "off-target" effects reducing economical resource and time. The druggability of GPCR protein families and critical roles played by them in complex diseases are explained. Class A, class B1, class C and class F are generally established family and GPCRs in phase I (19%), phase II(29%), phase III(52%) studies are also reviewed. From the phase I studies, frizzled receptors accounted for the highest in trial targets, neuropeptides in phase II and melanocortin in phase III studies. Also, the bioapplications for nanoparticles along with future prospects for both nanomedicine and GPCR drug industry are discussed. Further, the use of computational techniques and methods employed for different target validations are also reviewed along with their future potential for the GPCR based drug discovery.

Electrochemical detection of monosodium glutamate in foodstuffs based on Au@MoS2/chitosan modified glassy carbon electrode.

We report an amperometric immunosensor for the detection of monosodium glutamate (MSG) using a glassy carbon electrode modified with gold nanoparticle decorated on a molybdenum disulfide/chitosan (Au@MoS2/Ch) nanocomposite. In the present detection technique, Au@MoS2/Ch was used as a conductive matrix and anti-glutamate antibody was immobilized on to its surface via carbodiimide coupling method. Chemical and morphological attributes of the various components of the immunosensor were confirmed by UV-vis spectroscopy, SEM, TEM and XRD analysis. Electrochemical characterizations were carried out by CV, DPV and EIS. Overall results showed the effective fabrication of highly conductive Au@MoS2/Ch nanocomposite for sensitive electrochemical detection of MSG. A linear relationship was perceived between the change in current and concentration of MSG. The relationship was found to be consistent in the detection range of 0.05-200 µM. Statistical validation of the assay showed limit of detection and limit of quantification values as 0.03 and 0.1 µM, respectively (R2 = 0.99).

Chemical composition and antibacterial activity of essential oil from fruit of Micromelum integerrimum (Buch.-Ham. ex DC.) Wight & Arn. ex M. Roem.

The essential oil extracted from fruit of Micromelum integrrimum were evaluated through gas chromatography and gas chromatography-mass spectroscopy. 52 compounds were identified from the fruit oil representing 99.98% of the oil. The major components of the total fruit oil are monoterpene hydrocarbons (72.23%), oxygenated monoterpenes (14.78%) and sesquiterpene (11.54%) which were predominated by terpinolene (32.21%), α-pinene (17.24%), ß-pinene (17.24%), and camphene (4.05%). Moreover, other components that present in 1.45% were aromatic compounds, fatty acid, etc. The essential oil exhibited broad spectrum antimicrobial activity which is concentration dependent and 100 µL of the fruit oil showed the inhibition zones ranging from 7-16 mm. Fruit oil exhibited strong inhibition activity compared to standard anti-bacterial drug neomycin B (22 mm) against Bacillus subtilis MTCC 441 and Bacillus spizizenii ATCC 6633. This is the first hand report on the chemical profiles and promising anti-microbial activity of Micromelum integrrimum fruit essential oil towards Basillus Sp.

Blue-fluorescent and biocompatible carbon dots derived from abundant low-quality coals.

Coal is one of the most abundant natural carbonaceous materials. This paper reports a novel oxidative chemical method for the synthesis of high-value carbon dots (CDs) from cheap and abundant low-quality high­sulfur coals for use in high-end applications. These CDs were synthesized by using wet-chemical ultrasonic stimulation-induced process which is environmentally facile and less drastic compared to other chemical methods of production of CDs. The sizes of the synthesized CDs from different types of coal samples were estimated to be in the range of 1-4 nm, 1-6 nm, 2-5 nm, and 10-30 nm. The quantum yield (QY) of the CDs was determined and it was found to be around 3-14%. For high-end field application, the CDs were further tested for toxicity and were reported to be safe for environmental and biological applications. The cell image analysis under the fluorescence microscope further indicated that the synthesized CDs could be used as a promising bio-compatible material for optical-imaging as well as bio-imaging. The CDs showed promising fluorescent sensing property and can be utilized as a good probe for silver ion detection/sensing. The CDs is also found to be a promising reagent for silver nanoparticles synthesis. The results provide a new avenue for large-scale synthesis of CDs.

Volatile Inhibitors of Phosphatidylinositol-3-Kinase (PI3K) Pathway: Anticancer Potential of Aroma Compounds of Plant Essential Oils.

BACKGROUND: Cancer is a grave health problem for the world as the global cancer burden rises to 14 million new cases with 8.2 million deaths every year which is expected to rise by 70% in the next 2 decades as reported by the WHO.These steady rises in death demand for rapid developments in anti-cancer agents. Essential oils, being natural and multi-component complex systems have recently attracted a lot of attention in this search for novel anti-cancer agents. MATERIALS AND METHODS: The pharmaceutical attributes of essential oil components, specifically focusing on their affinity towards COX, 5-LOX, AKT, MDM2, PDK1 and mTOR which defines the phosphatidylinositol-3- kinase (PI3K) pathway, were assessed. 123 compounds present in essential oils of different plants were analyzed for their drug like attributes which were then allowed to dock with PI3K dependent receptors crucial for the development of cancer malignancies. Among them, 21 compounds were filtered possessing high druglikeness with favourable metabolism offered by major cytochromeP450 isoforms. Finally, the best docked compounds with highest binding affinities were employed for building a ligand based pharmacophore. Being inhibitors of P-glycoproteins, these molecules also exhibited good absorption profiles and noncarcinogenic properties. Further from these 21, six compounds were evaluated against A549 lung cancer cells. RESULTS: The pharmacophoric feature obtained can be applied for both designing and screening moieties for active inhibitors of the phosphatidylinositol-3-kinase pathway specifically from essential oil compounds and these final 21 compounds can be further promoted to studies for anti-cancer drug development. Among these, six compounds exhibited promising inhibitory results against A549 lung cancer cells. Furthermore, immunoblotting assay confirmed the efficacy of the compounds for inhibiting mTOR and AKT enzymes which are bandmasters for downstream signaling of thePI3K pathway. CONCLUSION: Methyl nonanoate, (R)-citronellol, cis-carveol (L-carveol), 3-methyl-Cyclohexanone, 4-carene and thujopsene were finally screened for PI3K targeted anti-cancer therapies which may find direct application as inhalers or sprays against lung cancer as these compounds are highly volatile.

Antimalarial silver and gold nanoparticles: Green synthesis, characterization and in vitro study.

In the present study, we are reporting antimalarial potential of silver (AgNPs) and gold (AuNPs) nanoparticles synthesized by leaf and bark extract of Syzygium jambos (L.) Alston (Myrtaceae). AuNPs and AgNPs obtained by both the extracts were characterized using UV-vis spectroscopy, zeta potential, transmission electron microscopy (TEM), X-ray diffraction (XRD), and Fourier Transform Infrared spectroscopy (FTIR). NMR and FTIR spectra indicate that the saccharides and phenolics present in the S. jambos extracts were the major contributors responsible for the synthesis and stabilization of NPs. NPs were also synthesized by chemical methods and were compared for their antiplasmodial potential against chloroquine sensitive (3D7) and resistant (Dd2) strain of Plasmodium falciparum by using 24h schizont maturation assay. AgNPs synthesized by both the extracts showed higher antiplasmodial activity than the rest. Further, NPs synthesized by S. jambos extracts have shown insignificant cytotoxicity against human cervical cancer cell line (HeLa) and rat skeletal muscle cell line (L6), which proved their biocompatibility.

Arsenic-induced instrumental genes of apoptotic signal amplification in death-survival interplay.

Arsenic is a global health concern at present and it is well reported for causing systemic toxicity. It is also well known for generation of free radical and inducing apoptosis in different cell types. Paradoxically arsenic is reported to be a susceptible carcinogen as well. There are several reports demonstrating diverse mechanism of apoptosis in different cell types. However, the universal scenario of instrumental genes and their interaction leading to amplification of apoptotic signal are yet to be completely uncovered, which is predicted here. Conventional studies on signaling pathway aided by time and concentration kinetics data are inadequate for prediction of anchored genes for apoptotic signal amplification. Therefore, expression profile-based approach is adopted. Core apoptosis related and glutathione metabolism genes in 1 and 10 µM of arsenic-treated HepG2 cells were analyzed after 12 h of incubation. An arsenic treatment of 1 µM exhibits no cell death at 12 h, whereas 10 µM arsenic treatment reveals around 50% cell death at 12 h. Results depict 28 and 44 affected genes in 1 and 10 µM arsenic-treated cells, respectively. Early initiation of apoptotic signaling is detected in no cell death regimens (at 1 µM), whereas amplified apoptotic signal is demonstrated at 50% cell death regimens (at 10 µM). Instrumental genes involved in progression of apoptosis in the concourse of cell death and survival is designated from the responsive genes common to both the condition. We predict the initiation process is fairly aided by the activation of intrinsic pathway, which is amplified via TNF signaling and extrinsic pathway. Furthermore, regulatory genes involved in interplay between apoptosis/anti-apoptosis and their interactions are demonstrated here.