Genome sequencing and assembly of Indian golden silkmoth, Antheraea assamensis Helfer (Saturniidae, Lepidoptera).

Muga silkworm (Antheraea assamensis), one of the economically important wild silkmoths, is unique among saturniid silkmoths. It is confined to the North-eastern part of India. Muga silk has the highest value among the other silks. Unlike other silkmoths, A. assamensis has a low chromosome number (n = 15), and ZZ/ZO sex chromosome system. Here, we report the first high-quality draft genome of A. assamensis, assembled by employing the Illumina and PacBio sequencing platforms. The assembled genome of A. assamensis is 501.18 Mb long, with 2697 scaffolds and an N50 of 683.23 Kb. The genome encompasses 18,385 protein-coding genes, 86.29% of which were functionally annotated. Phylogenetic analysis of A. assamensis revealed its divergence from other Antheraea species approximately 28.7 million years ago. Moreover, an investigation into detoxification-related gene families, CYP450, GST, and ABC-transporter, revealed a significant expansion in A. assamensis as compared to the Bombyx mori. This expansion is comparable to Spodoptera litura, suggesting adaptive responses linked to the polyphagous behavior observed in these insects. This study provides valuable insights into the molecular basis of evolutionary divergence and adaptations in muga silkmoth. The genome assembly reported in this study will significantly help in the functional genomics studies on A. assamensis and other Antheraea species along with comparative genomics analyses of Bombycoidea insects.

Molecular-iodine catalyzed selective construction of cyclopenta[b]indoles from indoles and acetone: a green gateway to indole-fused cycles.

Molecular-iodine catalyzed access to an important class of bio-relevant indole derivatives, cyclopenta[b]indoles, has been achieved via a cascade addition/intramolecular cyclization reaction of indoles and acetone. Explorations of diverse substitution patterns revealed an essential substrate-control in the reaction. The high-density electronic core of indole is pivotal in favouring the formation of indolyl-cyclopenta[b]indole derivatives; in contrast, the electron deficiency of the core hindered the cyclization process, directing the formation of bis(indolyl)propanes. Investigations on the mechanistic pathway revealed that bis(indolyl)alkanes were the intermediates for the addition-cyclization process. This simple experimental method provides sustainable synthetic access to cyclopentannulated indoles.

Halogen bonding assisted site-selective C-3 triaryl methylation of indoles and N-triaryl methylation of imidazoles.

Halogen bonding triggered by the Lewis basic nature of acetonitrile catalyzes the site-selective C-3 triaryl methylation of indoles and N-triaryl methylation of imidazoles with trityl chlorides under catalyst-, metal-, and additive-free conditions at room temperature. This method generates a quaternary carbon centre appended to a heterocyclic moiety. UV-Vis and FT-IR analyses indicate the existence of halogen bonding which is the driving force of the reaction. This approach is suitable for a wide range of substrates, furnishing moderate to excellent yields (up to 100%) of triaryl methylated products under ambient reaction conditions. Equimolar amounts of reactants are sufficient to obtain the optimum yield and in some cases pure products can be obtained without column chromatography.

N,N'-Dimethylurea as an efficient ligand for the synthesis of pharma-relevant motifs through Chan-Lam cross-coupling strategy.

N,N'-Dimethylurea (DMU) is introduced as a ligand to aid the Chan-Lam N-arylation of primary amides, amines, and 3-aminophenols with arylboronic acids and its ester derivative as the arylating associate. The developed methodology is catalyzed by Cu and its in situ complexation with DMU brings about efficient synthesis of N-arylated anilines, 3-aminophenols, and primary amides in moderate to good yields (50-90%). The [Cu2(OAc)4(DMU)2] complex is synthesized and characterized by single crystal structure elucidation. The catalyst is cheap, free from prior synthesis of a metal complex, provides chemoselectivity towards the N-arylation of 3-aminophenols, and is suitable for mono-arylation of primary amides. The synthetic utility of the methodology is tested in the post-modification of two active pharmaceutical ingredients (APIs). The developed catalytic system extends the scope of N,N'-dimethylurea as an auxiliary in inexpensive and versatile Cu catalysis.

Revisiting the synthesis of aryl nitriles: a pivotal role of CAN.

Facilitated by the dual role of Ceric Ammonium Nitrate (CAN), herein we report a cost-effective approach for the cyanation of aryl iodides/bromides with CAN-DMF as an addition to the existing pool of combined cyanation sources. In addition to being an oxidant, CAN acts as a source of nitrogen in our protocol. The reaction is catalyzed by a readily available Cu(ii) salt and the ability of CAN to generate ammonia in the reaction medium is utilized to eliminate the additional requirement of a nitrogen source, ligand, additive or toxic reagents. The mechanistic study suggests an evolution of CN- leading to the synthesis of a variety of aryl nitriles in moderate to good yields. The proposed mechanism is supported by a series of control reactions and labeling experiments.

RNAinsecta: A tool for prediction of precursor microRNA in insects and search for their target in the model organism Drosophila melanogaster.

INTRODUCTION AND BACKGROUND: Pre-MicroRNAs are the hairpin loops from which microRNAs are produced that have been found to negatively regulate gene expression in several organisms. In insects, microRNAs participate in several biological processes including metamorphosis, reproduction, immune response, etc. Numerous tools have been designed in recent years to predict novel pre-microRNA using binary machine learning classifiers where prediction models are trained with true and pseudo pre-microRNA hairpin loops. Currently, there are no existing tool that is exclusively designed for insect pre-microRNA detection. AIM: Application of machine learning algorithms to develop an open source tool for prediction of novel precursor microRNA in insects and search for their miRNA targets in the model insect organism, Drosophila melanogaster. METHODS: Machine learning algorithms such as Random Forest, Support Vector Machine, Logistic Regression and K-Nearest Neighbours were used to train insect true and false pre-microRNA features with 10-fold Cross Validation on SMOTE and Near-Miss datasets. miRNA targets IDs were collected from miRTarbase and their corresponding transcripts were collected from FlyBase. We used miRanda algorithm for the target searching. RESULTS: In our experiment, SMOTE performed significantly better than Near-Miss for which it was used for modelling. We kept the best performing parameters after obtaining initial mean accuracy scores >90% of Cross Validation. The trained models on Support Vector Machine achieved accuracy of 92.19% while the Random Forest attained an accuracy of 80.28% on our validation dataset. These models are hosted online as web application called RNAinsecta. Further, searching target for the predicted pre-microRNA in Drosophila melanogaster has been provided in RNAinsecta.

Towards integrated sustainable biofuel and chemical production: An application of banana pseudostem ash in the production of biodiesel and recovery of lignin from bamboo leaves.

This study investigated an integrated approach to the biowaste transformation and valorization of byproducts. Biochar obtained from the banana pseudostem was calcined to synthesize a heterogeneous catalyst and sustainably prepare a highly alkaline solution. The ash was utilized directly as a heterogeneous catalyst in biodiesel production from waste cooking oil. At the same time, an alkaline solution prepared from the ash was used for delignification and recovery of lignin from bamboo leaves by the hydrothermal reaction. Techniques like Fourier-transform infrared spectroscopy (FTIR), Field emission scanning electron microscopy (FESEM), Brunauer-Emmett-Teller (BET), Transmission electron microscopy (TEM), and Energy dispersive X-ray (EDX) were applied to characterized the catalyst. The alkaline solution was analyzed with Atomic absorption spectroscopy (AAS). The Response surface methodology (RSM) technique was considered for the optimization of different parameters in the transesterification and hydrothermal reaction. Under the optimized condition, waste cooking oil (WCO) to Fatty acid methyl ester (FAME) conversion was 97.56 ± 0.11%, and lignin recovery was 43.20 ± 0.45%. While at the best operating pyrolysis temperature, the liquid fraction yield from the banana pseudostem (500 °C) was 38.10 ± 0.31 wt%. This integrated study approach encourages the inexpensive, sustainable, and environment-friendly pathway for synthesizing catalysts and preparing a highly alkaline solution for the valorization of biowaste into biofuel and biochemicals.

Papaya latex mediated synthesis of prism shaped proteolytic gold nanozymes.

Beyond natural enzymes, the artificially synthesized nanozymes have attracted a significant interest as it can overcome the limitations of the former. Here, we report synthesis of shape controlled nanozymes showing proteolytic activity using Carica papaya L. (papaya) latex. The nanozymes synthesized under optimized reaction conditions exhibited sharp SPR peak around 550 nm with high abundance (45.85%) of prism shaped particles. FTIR analysis and coagulation test indicated the presence of papaya latex enzymes as capping agents over the gold nanoprisms. The milk clot assay and the inhibition test with egg white confirmed the proteolytic activity of the nanozymes and the presence of cysteine protease on it, respectively. The nanozymes were found to be biocompatible and did not elicit any toxic response in both in-vitro and in-vivo study. Based on our findings, we envisage that these biocompatible, shape-specific nanozymes can have potential theragnostic applications.

Microwave-mediated enzyme-linked immunosorbent assay procedure.

Here we demonstrate a novel microwave-mediated enzyme-linked immunosorbent assay (MELISA) method that has dramatically reduced the enzyme-linked immunosorbent assay (ELISA) timing to less than 5 min with a result comparable to that obtained by 18-h conventional ELISA. Efficacy of the MELISA procedure is demonstrated by detecting human immunoglobulin G (IgG), rabbit IgG, human immunoglobulin E (IgE), human interleuken 1ß (IL-1ß), Entamoeba histolytica antibody, and Aspergillus fumigatus antibody. MELISA could be an excellent substitute for time-consuming conventional ELISA for rapid diagnosis of diseases in cases of medical urgency, outbreak of infectious diseases, and screening of samples in blood banks or emigration counters.

Antioxidant, antibacterial, cytotoxic, and apoptotic activity of stem bark extracts of Cephalotaxus griffithii Hook. f.

BACKGROUND: Cephalotaxus spp. are known to possess various therapeutic potentials. Cephalotaxus griffithii, however, has not been evaluated for its biological potential. The reason may be the remoteness and inaccessibility of the habitat where it is distributed. The main aim of this study was to: (1) evaluate multiple biological potentials of stem bark of C. griffithii, and (2) identify solvent extract of stem bark of C. griffithii to find the one with the highest specific biological activity. METHODS: Dried powder of stem bark of C. griffithii was exhaustively extracted serially by soaking in petroleum ether, acetone and methanol to fractionate the chemical constituents into individual fractions or extracts. The extracts were tested for total phenolic and flavonoid content, antioxidant (DPPH radical scavenging, superoxide radical scavenging, and reducing power models), antibacterial (disc diffusion assay on six bacterial strains), cytotoxic (MTT assay on HeLa cells), and apoptotic activity (fluorescence microscopy, DNA fragmentation assay, and flow cytometry on HeLa cells). RESULTS: Among the three extracts of stem bark of C. griffithii, the acetone extract contained the highest amount of total phenolics and flavonoids and showed maximum antioxidant, antibacterial, cytotoxic (IC50 of 35.5 ± 0.6 µg/ml; P < 0.05), and apoptotic (46.3 ± 3.6% sub-G0/G1 population; P < 0.05) activity, followed by the methanol and petroleum ether extracts. However, there was no significant difference observed in IC50 values (DPPH scavenging assay) of the acetone and methanol extracts and the positive control (ascorbic acid). In contrast, superoxide radical scavenging assay-based antioxidant activity (IC50) of the acetone and methanol extracts was significantly lower than the positive control (P < 0.05). Correlation analysis suggested that phenolic and flavonoid content present in stem bark of C. griffithii extracts was responsible for the high antioxidant, cytotoxic, and apoptotic activity (P < 0.05). CONCLUSIONS: Stem bark of C. griffithii has multiple biological effects. These results call for further chemical characterization of acetone extract of stem bark of C. griffithii for specific bioactivity.

Mitogenome-wise codon usage pattern from comparative analysis of the first mitogenome of Blepharipa sp. (Muga uzifly) with other Oestroid flies.

Uziflies (Family: Tachinidae) are dipteran endoparasites of sericigenous insects which cause major economic loss in the silk industry globally. Here, we are presenting the first full mitogenome of Blepharipa sp. (Acc: KY644698, 15,080 bp, A + T = 78.41%), a dipteran parasitoid of Muga silkworm (Antheraea assamensis) found in the Indian states of Assam and Meghalaya. This study has confirmed that Blepharipa sp. mitogenome gene content and arrangement is similar to other Tachinidae and Sarcophagidae flies of Oestroidea superfamily, typical of ancestral Diptera. Although, Calliphoridae and Oestridae flies have undergone tRNA translocation and insertion, forming unique intergenic spacers (IGS) and overlapping regions (OL) and a few of them (IGS, OL) have been conserved across Oestroidea flies. The Tachinidae mitogenomes exhibit more AT content and AT biased codons in their protein-coding genes (PCGs) than the Oestroidea counterpart. About 92.07% of all (3722) codons in PCGs of this new species have A/T in their 3rd codon position. The high proportion of AT and repeats in the control region (CR) affects sequence coverage, resulting in a short CR (Blepharipa sp.: 168 bp) and a smaller tachinid mitogenome. Our research unveils those genes with a high AT content had a reduced effective number of codons, leading to high codon usage bias. The neutrality test shows that natural selection has a stronger influence on codon usage bias than directed mutational pressure. This study also reveals that longer PCGs (e.g., nad5, cox1) have a higher codon usage bias than shorter PCGs (e.g., atp8, nad4l). The divergence rates increase nonlinearly as AT content at the 3rd codon position increases and higher rate of synonymous divergence than nonsynonymous divergence causes strong purifying selection. The phylogenetic analysis explains that Blepharipa sp. is well suited in the family of insectivorous tachinid maggots. It's possible that biased codon usage in the Tachinidae family reduces the effective number of codons, and purifying selection retains the core functions in their mitogenome, which could help with efficient metabolism in their endo-parasitic life style and survival strategy.

Green synthesis of gold nanoparticles using Nyctanthes arbortristis flower extract.

The present study explores the reducing and capping potentials of ethanolic flower extract of the plant Nyctanthes arbortristis for the synthesis of gold nanoparticles. The extract at different volume fractions were stirred with HAuCl4 aqueous solution at 80 °C for 30 min. The UV-Vis spectroscopic analysis of the reaction products confirmed successful reduction of Au(3+) ions to gold nanoparticles. Transmission electron microscope (TEM) revealed dominant spherical morphology of the gold nanoparticles with an average diameter of 19.8 ± 5.0 nm. X-ray diffraction (XRD) study confirmed crystalline nature of the synthesized particles. Fourier transform infra-red (FTIR) and nuclear magnetic resonance (NMR) analysis of the purified and lyophilized gold nanoparticles confirmed the surface adsorption of biomolecules during preparation and caused long-term (6 months) stability. Low reaction temperature (25 °C) favored anisotropy. The strong reducing power of the flower extract can also be tested in the green synthesis of other metallic nanoparticles.

Encapsulation of curcumin in alginate-chitosan-pluronic composite nanoparticles for delivery to cancer cells.

We report a nanoformulation of curcumin with a tripolymeric composite for delivery to cancer cells. The composite nanoparticles (NPs) were prepared by using three biocompatible polymers-alginate (ALG), chitosan (CS), and pluronic-by ionotropic pre-gelation followed by polycationic cross-linking. Pluronic F127 was used to enhance the solubility of curcumin in the ALG-CS NPs. Atomic force and scanning electron microscopic analysis showed that the particles were nearly spherical in shape with an average size of 100 +/- 20 nm. Fourier transform-infrared analysis revealed potential interactions among the constituents in the composite NPs. Encapsulation efficiency (%) of curcumin in composite NPs showed considerable increase over ALG-CS NPs without pluronic. The in vitro drug release profile along with release kinetics and mechanism from the composite NPs were studied under simulated physiological conditions for different incubation periods. A cytotoxicity assay showed that composite NPs at a concentration of 500 microg/mL were nontoxic to HeLa cells. Cellular internalization of curcumin-loaded composite NPs was confirmed from green fluorescence inside the HeLa cells. The half-maximal inhibitory concentrations for free curcumin and encapsulated curcumin were found to be 13.28 and 14.34 muM, respectively. FROM THE CLINICAL EDITOR: A nanoformulation of curcumin with a tri-component polymeric composite for delivery to cancer cells is reported in this paper. Cellular internalization of curcumin loaded composite nanoparticles was confirmed from green fluorescence inside the HeLa cells.

Towards sustainable biodiesel and chemical production: Multifunctional use of heterogeneous catalyst from littered Tectona grandis leaves.

Waste biomass derived heterogeneous catalyst is an excellent alternative to chemically synthesized catalysts. In this work, calcined Tectona grandis leaves were proposed as an eco-friendly, renewable and low cost heterogeneous base catalyst. The prepared catalyst was examined by FTIR, XRD, XPS, SEM, EDX, TEM, TGA, BET and Hammett indicator test. The catalyst has an appealing nature towards various chemical transformations due to its basic surface sites provided by alkali and alkaline earth metals. The efficiency of the catalyst was successfully investigated by its application in biodiesel production. The products were confirmed by 1H and 13C NMR. 100% FAME conversion was attained using a catalyst loading of 2.5 wt% under optimized reaction parameters. The catalyst was further explored for Knoevenagel condensation reaction, in which it showed its effectiveness and recyclability towards the formation of benzylidenemalononitrile derivatives of aryl aldehydes. Thus, it is a potential 'green catalyst' derived from waste biomass without any addition of chemicals that can replace the industrial base catalysts used for biodiesel production and Knoevenagel reaction and makes the protocol environmentally benign.

Molecular Iodine-Catalyzed Selective C-3 Benzylation of Indoles with Benzylic Alcohols: A Greener Approach toward Benzylated Indoles.

Iodine-catalyzed selective C-3 benzylation of indoles with benzylic alcohols is developed. The reaction proceeds with molecular iodine as the catalyst under ligand-, metal-, and base-free conditions and tolerates wide functionalities. The experimental observations account for the halogen-bond activation mechanistic pathway for the molecular iodine catalysis.

Effect of Substrates on Catalytic Activity of Biogenic Palladium Nanoparticles in C-C Cross-Coupling Reactions.

This work describes a practical methodology for C-C bond formation reactions with the aid of biogenic palladium nanoparticles, which are synthesized by using phytochemicals extracted from two common plant species. Comparative studies have been done on the activity of two plant species (Ocimum sanctum and Aloe vera) in generation of palladium nanoparticles via ex situ and in situ methods. The structural and morphological characteristics of the nanoparticles are examined by UV/visible spectroscopy, powder X-ray diffraction, X-ray photoelectron spectroscopy, Fourier transform infrared spectroscopy, and transmission electron microscopy analyses. We have observed a significant influence of the substrates on the catalytic activity of the palladium nanoparticles in Sonogashira and Suzuki cross-coupling reactions.

Fluorescence study of the curcumin-casein micelle complexation and its application as a drug nanocarrier to cancer cells.

In milk caseins exists a natural nanostructure, which can be exploited as a carrier of hydrophobic drugs. Here we investigated the complex formation of curcumin with bovine casein micelles (CMs) and its use as a vehicle for drug delivery to cancer cells. DLS studies of the CM suspension that was stable in buffer solution (pH 7.4) showed an average size distribution of <200 nm. SEM and AFM studies showed that the particles were roughly spherical in shape. Steady-state fluorescence spectroscopy of the CM-curcumin complex formation revealed that curcumin molecules formed complexes with CMs (CM-curcumin complex) through hydrophobic interactions. The binding constant for the CM-curcumin interaction was calculated to be 1.48 x 10(4) M(-1), as determined by the curcumin fluorescence. Fluorescence quenching showed that curcumin molecules quench the intrinsic fluorescence of caseins upon binding. We evaluated the utility of CMs as carriers of curcumin by using in vitro cultured HeLa cells. Cytotoxicity studies of HeLa cells revealed that the IC50 of free curcumin and the CM-curcumin complex was 14.85 and 12.69 microM, respectively.

Phospholipases play multiple cellular roles including growth, stress tolerance, sexual development, and virulence in fungi.

Phospholipases are ubiquitous enzymes that hydrolyze phospholipids. Based on the cleavage site of the ester linkage in the substrate phospholipids, phospholipases are classified into four major types, phospholipase A (PLA), phospholipase B (PLB), phospholipase C (PLC), and phospholipase D (PLD), which are further classified into various subtypes. Phospholipases hydrolyze phospholipids into various signaling products including phosphatidic acid (PA), diacylglycerol (DAG), free fatty acids (FFAs), and lyso-phospholipids (LPLs). These signaling products regulate numerous processes such as cytoskeletal dynamics, growth, homeostasis, membrane remodeling, nutrient acquisition, secretion, signal transduction, stress tolerance, sexual development, and virulence in various organisms including fungi. Due to these key cellular roles, phospholipases are also promising targets in diagnostic and therapeutic applications. In this review, we discuss current knowledge about the cellular roles of different classes of phospholipases in fungi.

Greener Biogenic Approach for the Synthesis of Palladium Nanoparticles Using Papaya Peel: An Eco-Friendly Catalyst for C-C Coupling Reaction.

The development of a green and sustainable synthetic methodology still remains a challenge across the globe. Encouraging the prevailing challenge, herein, we have synthesized Pd nanoparticles (Pd NPs) in a green and environmentally viable route, using the extract of waste papaya peel without the assistance of any reducing agents, high-temperature calcination, and reduction procedures. The biomolecules present in the waste papaya peel extract reduced Pd(II) to nanosize Pd(0) in a one-pot green and sustainable process. As a catalyst, the new Pd NPs offer a simple and efficient methodology in direct Suzuki-Miyaura and Sonogashira coupling with excellent yields under mild reaction conditions.

The mitochondrial genome of Muga silkworm (Antheraea assamensis) and its comparative analysis with other lepidopteran insects.

Muga (Antheraea assamensis) is an economically important silkmoth endemic to the states of Assam and Meghalaya in India and is the producer of the strongest known commercial silk. However, there is a scarcity of genomic and proteomic data for understanding the organism at a molecular level. Our present study is on decoding the complete mitochondrial genome (mitogenome) of A. assamensis using next generation sequencing technology and comparing it with other available lepidopteran mitogenomes. Mitogenome of A. assamensis is an AT rich circular molecule of 15,272 bp (A+T content ~80.2%). It contains 37 genes comprising of 13 protein coding genes (PCGs), 22 tRNA and 2 rRNA genes along with a 328 bp long control region. Its typical tRNAMet-tRNAIle-tRNAGln arrangement differed from ancestral insects (tRNAIle-tRNAGln-tRNAMet). Two PCGs cox1 and cox2 were found to have CGA and GTG as start codons, respectively as reported in some lepidopterans. Interestingly, nad4l gene showed higher transversion mutations at intra-species than inter-species level. All PCGs evolved under strong purifying selection with highest evolutionary rates observed for atp8 gene while lowest for cox1 gene. We observed the typical clover-leaf shaped secondary structures of tRNAs with a few exceptions in case of tRNASer1 and tRNATyr where stable DHU and TΨC loop were absent. A significant number of mismatches (35) were found to spread over 19 tRNA structures. The control region of mitogenome contained a six bp (CTTAGA/G) deletion atypical of other Antheraea species and lacked tandem repeats. Phylogenetic position of A. assamensis was consistent with the traditional taxonomic classification of Saturniidae. The complete annotated mitogenome is available in GenBank (Accession No. KU379695). To the best of our knowledge, this is the first report on complete mitogenome of A. assamensis.