Browsing by Author "Bhushan, Satej"
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Item Caulerpa taxifolia inhibits cell proliferation and induces oxidative stress in breast cancer cells(Springer, 2018) Mehra, Richa; Bhushan, Satej; Yadav, Umesh Prasad; Bast, Felix; Singh, SandeepCaulerpa taxifolia (M. Vahl) C. Agardh or killer alga is known to possess several bioactive secondary metabolites with unique structural modifications. We investigated anti-oxidant and anti-proliferative activity of C. taxifolia extract (CTE) on breast and lung cancer cells, along with possible effects on mitochondrial membrane potential (MMP) and cell cycle progression. The results revealed up to 6-folds increase in reactive oxygen species (ROS), 2-folds increase in glutathione reductase (GR) activity, 1.7-fold increase in superoxide dismutase (SOD) activity and 1.8-fold change in catalase activity w.r.t. untreated cells i.e. 10.72 to 21.44 nmol/min/mL, 2.0 to 3.49 U/mL and 37.51 to 69.26 U/min/g FW, respectively, in MDA-MB-cells. Likewise, selective anti-proliferative activity with IC50 0.19 + 0.1, 0.27 + 0.1, and 0.43 + 0.1 μg/μL, was recorded in MDA-MB-231, T-47D, and H1299 cells. In addition, dose-dependent increase in MMP of up to 40% and G1/S phase mitotic arrest was documented by CTE treatment in MDA-MB-231 cells. The results suggest an anti-proliferative and oxidative stress inducing activity of CTE. Changes in MMP and cell cycle arrest further support the anti-cancer effects of CTE. It is believed that C. taxifolia may be considered as a potent source of anti-cancer drugs, subject to further validations.Item Cladophora goensis sp. nov. (Cladophorales, Ulvophyceae) –a bloom forming marine algae from Goa, India(CSIR- NISCAIR, 2015) Bast, Felix; John, Aijaz Ahmad; Bhushan, SatejA new species of green seaweed, Cladophora goensis sp. nov. (Cladophorales, Ulvophyceae), had been discovered from Vasco-da-Gamma, Goa, India. This species formed algal bloom of moderate intensity in the Bay of Mormugao, on the west coast of India. Observations suggest combination of a number of morphological characteristics of this alga distinct from previously described members of this genus, including parietal chloroplast surrounding central hollow and bilenticular pyrenoids. Molecular sequence data at Nuclear ribosomal DNA Internal Transcribed Spacer-1 and 2 (ITS1 and ITS2) regions along with intervening 5.8S rRNA indicated Kimura-2-Parameter (T3P) pair-wise distance of 1.77 x 10-1 between this species and the nearest phylogenetic accession of Cladophora glomerata. In phylogenetic reconstructions using Bayesian Inference and Maximum Likelihood, this species was not part of any monophyletic clades comprising any of the previously described species of this genus at the locus studied, thereby ascertaining conformity with phylogenetic species concept. With this discovery, a new phylogenetically primitive morphological synapomorphy of “pseudo dichotomous profuse branching” has been revealed for cladophoralean algae, and this is the single most important morphological characteristic of this bloom- forming seaweed.Item Cladophora goensis- a bloom forming marine algae from Goa, India.(2015) Bast, Felix; John,Aijaz A.; Bhushan, SatejA new species of green seaweed, Cladophora goensis sp. nov. (Cladophorales, Ulvophyceae), had been discovered in our last field trip to Vasco-da-Gamma, Goa, India. This species formed algal bloom of moderate intensity in the Bay of Mormugao, on the west coast of India. Our observations suggest combination of a number of morphological characteristics of this alga distinct from previously described members of this genus, including parietal chloroplast surrounding central hollow and bilenticular pyrenoids. Molecular sequence data at Nuclear ribosomal DNA Internal Transcribed Spacer-1 and 2 (ITS1 and ITS2) regions along with intervening 5.8S rRNA indicated Kimura-2-Parameter (K2P) pair-wise distance of 1.77 x 10-1 between this species and the nearest phylogenetic accession of Cladophora glomerata. In our phylogenetic reconstructions using Bayesian Inference and Maximum Likelihood, this species was not part of any monophyletic clades comprising any of the previously described species of this genus at the locus studied, thereby ascertaining conformity with phylogenetic species concept. With this discovery, a new phylogenetically primitive morphological synapomorphy of “pseudodichotomous profuse branching” has been revealed for cladophoralean algae, and we argue that this is the single most important morphological characteristic of this bloom-forming seaweed.Item Cystathionine β-Lyase-Like Protein with Pyridoxal Binding Domain Characterized in Leishmania major by Comparative Sequence Analysis and Homology Modelling(Hindawi, 2013) Negi, Arvind; Bhushan, Satej; Gupta, Pawan; Garg, Prabha; Kumar, RajCystathionine β-lyase-like protein (CBLP), one of the key enzymes involved in methionine biosynthesis utilising pyridoxal phosphate (PLP) as a cofactor, has recently been reported in Leishmania major. Its presence in the parasite and absence in humans warrant its full characterisation and fruition as a potent, selective, and inevitable druggable target. Due to the unavailability of X-ray 3D structure of CBLP, a homology model for this protein was developed for the first time. The model was evaluated for PLP binding site and various conserve domain residues of the protein recommended by comparative sequence analyses by different protein analysis tools. The model was validated and discovered to be robust and statistically significant. The final model was superimposed on template of Arabidopsis thaliana (PDB ID: 1IBJ) and RMSD was found to be 0.486. The PLP binding site residues of both the proteins were ensued to be highly conserved indicated by Gly71, Met72, Tyr95, Asp169, and Ser193 as well as formation of aldimine bond with Lys194. This was further verified through molecular simulation of PLP into the cofactor binding site of the modelled protein. The present study may therefore play a directing role in the designing of novel, potential, and selective antileishmanial agents.Item DB IndAlgae: An on-line resource of marine algae from India identified on the basis of molecular and morphological features(Indian Academy of Sciences, 2016) Bhushan, Satej; Mehra, Richa; Rani, Pooja; Bast, FelixDbIndAlgae is a free on-line database of marine algae from India. It provides information about the geographical distribution, morphological characteristics and most importantly, sequence data of marine algae collected from different coasts. It is the only database which contains molecular data of the algal species from India. Identification of the species is based on both morphological as well as molecular information. The database also serves as an interface to the herbarium maintained at the Centre for Plant Sciences, Central University of Punjab, Bathinda. So far the database lists 45 marine algal species. Some algal species have been identified for the first time from India, but have already been reported from places other than India, whereas new species are those which have been identified by us for the first time as a part of this study and have never been reported elsewhere.Item DNA barcode-based identification and comparative anti-cancer effects of different species of brown seaweed Sargassum C. Agardh of Indian coasts(Central University of Punjab, 2018) Bhushan, Satej; Bast, Felix & Singh, SandeepSargassum C. Agardh is a ubiquitous, multicellular brown seaweed that represents the most species-rich genus of the brown algal order Fucales, with more than 500 species reported worldwide. The present study aimed to identify different Sargassum isolates from India by DNA barcoding of mitochondrial (cox3), chloroplast (rbcL), and nuclear (18S) regions and further phylogenetic analyses. Total of 17 geographical isolates were collected across Indian coasts. Phylogeny reconstruction using Bayesian Inference was done which suggested congruency with known taxonomic hierarchy of Sargassum. Total of five different species were identified (S. portierianum, S. cymosum, S.aquifolium, S. ilicifolium, S. polycystum). In addition, comparative evaluation of anti-cancer potential of all the isolates was carried out and putative relationship between phylogeny and anticancer potential was established. MTT assay with 3 different cell lines showed cytotoxicity with IC50 as low as 0.167 ± 0.01, 0.243 ± 0.007, 0.25 ± 0.03 µg/µL in MDA-MB-231 (Breast Cancer), T-47D (Breast Cancer), H1299 (Lung Cancer) cells respectively, while no toxicity was observed with human peripheral blood mononuclear cells (hPBMCs). I was also able to isolate one lead aliphatic compound (SA1) whch was identified to be a polysaccharide using NMR spectroscopy. Similar to the extract, purified compound SA1 also showed anticancer activity. Further evaluations revealed that SA1 as well as the extracts interfere with the antioxidant defence components of cancer cells (SOD, Catalase, and GR) which results in the induction of mitochondrial death pathway at G1 phase (for extracts) as well as at G2M phase (for SA1). Extracts as well as SA1 were also able to inhibit cancer cell migration at sub IC50 doses. In addition, sub IC50 treatments lead to decreased colony formation compared to the control. Overall, our results show that these extracts as well as SA1 are able to target multiple properties of cancerItem DNA Barcoding and phylogography of brown seaweeds of coasts of indian subcontinent(Central University of Punjab, 2013) Bhushan, Satej; Bast, FelixAlgae are one of the diverse groups of ubiquitous autotrophs. Their use as food was more or less initially confined to few East Asian and South American countries like China, Japan, Korea, Chile, Argentina and Brazil, but with the increased demand and limited supply for food combined with the medicinal properties of the marine macroalgae, they started getting the attention of policy makers and researchers alike all around the world. Brown seaweeds (Phaeophyceae) are mostly marine and characterized by presence of pigment fucoxanthin which gives them its coloured appearance. The present work aims to provide detailed molecular analysis of the brown seaweeds found in Indian coastal regions to study and characterize it taxonomically which has not been done till now in Indian context. Out of all the samples processed, one invasive species was detected, Sargassum zhangii, which is the first report of this algal species outside Chinese waters. The conspecificity was confirmed by a multi-faceted approach, including comparative morphology, microscopy, genetic distance analysis and computational phylogenetics using Maximum Likelihood and Bayesian Inference methods.Item DNA Barcoding of a new record of epi-endophytic green algaeUlvellaleptochaete(Ulvellaceae, Chlorophyta) in India(Indian Academy of Science, 2014) Bast, Felix; Bhushan, Satej; John, Aijaz AhmadEpi-endophytic green algae comprise one of the most diverse and phylogenetically primitive groups of green algae and are considered to be ubiquitous in the world's oceans; however, no reports of these algae exist from India. Here we report the serendipitous discovery of Ulvella growing on intertidal green algae Cladophora glomerata and benthic red algae Laurencia obtusa collected from India. DNA barcodes at nuclear ribosomal DNA Internal Transcriber Spacer (nrDNA ITS) 1 and 2 regions for Indian isolates from the west and east coasts have been generated for the first time. Based on morphology and DNA barcoding, isolates were identified as Ulvella leptochaete. Phylogenetic reconstruction of concatenated dataset using Maximum Likelihood method differentiated Indian isolates from other accessions of this alga available in Genbank, albeit with low bootstrap support. Monophyly of Ulvella leptochaete was obvious in both of our phylogenetic analyses. With this first report of epi-endophytic algae from Indian territorial waters, the dire need to catalogue its cryptic diversity is highlighted and avenues of future research are discussed.Item European Species of Subaerial Green Alga Trentepohlia annulata (Trentepohliales, Ulvophyceae) Caused Blood Rain in Kerala, India(OMICS, 2015) Bast, Felix; Bhushan, Satej; John, Aijaz Ahmad; Achankunju, Jackson; MV Panikkar, Nadaraja; Hametner, Christina; Stocker-W�rg�tter, ElfriedeIn 2011, isolated parts in south Indian state of Kerala as well as neighboring Sri Lanka experienced sporadic spell of red colored “blood rain”, cause of which was later attributed to terrestrial subaerial microalgae of the genus Trentepohlia. Green algae of this genus is commonly found living in symbiosis as phycobionts of lichens in the freeliving form in adjoining tropical rain forests, however, specific identity of which have never been determined. It is known that lichens disperse small algal-hyphal packages, so-called soredia, for vegetative reproduction, which can explain the content of Trentepohlia-spores in the water. Given its extraordinary spore-dispersal mechanism via rain, we were specifically interested to know species-level identity of a randomly collected Trentepohlialean algae of “blood rain” region, and to investigate the possibility that this might had been introduced from elsewhere through areal route. Results of our comparative morphology and molecular phylogenetic analyses using the nuclear ribosomal DNA Internal Transcribed Spacers region concluded that this alga is Trentepohlia annulata – a species of which no previous reports exist from India. These two geographical isolates were separated by Kimura-2-Parameter pair-wise distance of 0.06-which in turn indicate a low rate of evolution at these loci that are renowned for rapid molecular evolution, suggestive of a recent introduction. Phylogeny reconstruction using Bayesian Inference and Maximum Likelihood methods resulted in well-resolved phylograms with a robust clade composed of these isolates. This finding indicates the existence of areal dispersal of algal spores on continental and global scales through “clouds over oceans”- a phenomenon earlier reported for bacteria and fungi, but for the first time in algae.Item Molecular assessment of invasive Carrageenophyte Kappaphycus alvarezii from India based on ITS-1 sequences(Taylor and Francis Ltd., 2016) Bast, Felix; John, Aijaz Ahmad; Bhushan, SatejThis is the first report on the molecular systematic characterization of the invasive red alga Kappaphycus alvarezii in India. Cultivated for the production of carrageenan around the Gulf of Mannar, southeast India, this exotic alga is causing havoc in coastal ecosystems by invading the habitats of a number of endemic species, especially soft corals. We have sequenced the nuclear ribosomal DNA internal transcribed spacer 1 barcode of this introduced species. In our phylogenetic analyses using Bayesian inference and maximum likelihood, this alga clustered within a well-supported clade of K.?alvarezii of mixed geographic origin. Phylogenetic analyses were congruent in their separation of K.?alvarezii and Kappaphycus striatum. Analyses also suggest that this species might have originated in the Malay Archipelago and spread elsewhere. Our results provide insights into the evolutionary and biogeographic patterns of this alga around the world. ? 2016 Dipartimento di Biologia, Universit? di Firenze.Item Morphological and molecular assessment of native carrageenophyte Hypnea valentiae (Cystocloniaceae, Gigartinales) in Indian Subcontinent(Phycological Society, 2014) Bast, Felix; Bhushan, Satej; John, Aijaz AhmadHypnea valentiae is an important red alga commercially cultivated in various parts of the world for the production of carrageenan. Presented in this report is findings of morphological and molecular observations of naturally-occurring populations of this alga collected from west and east coasts of India. Both the isolates had similar external as well as microscopic morphology. Nuclear ribosomal DNA Internal Transcribed Spacer-1 (ITS1) sequences from these geographical isolates had 4.35 x 10 -1 Tamura-3-Parameter (T3P) pairwise distance between them, which indicate significant evolutionary differences accumulated over time. In comparison, T3P distance between related genera Kappaphycus and Eucheuma was 1.85 x 10 -1 . In our phylogeny reconstruction using Bayesian Inference, both the isolates formed a well- supported clade along with the only available accession of this genus at ITS1 locus, indicating affiliation of both the isolates in this genus. Interestingly, isolate from the west coast was more basal in the phylogram, which suggests phylogenetically primitive position of this population. Newly generated DNA barcodes of the geographic isolates of this native carragenophyte in this study is expected to be a key in tracing its further dispersal routes, either natural or deliberate. This is the first report on the comparative morphological and molecular assessment of Hypnea from India.Item New record of Sargassum zhangii (Sargassaceae, Fucales) in India based on nuclear and mitochondrial DNA barcodes(Taylor and Francis Ltd., 2016) Bast, Felix; Bhushan, Satej; Rani, Pooja; John, Aijaz AhmadBrown algal genus Sargassum encompasses some of the most invasive seaweeds in the oceans across the world. Here we report the occurrence of Sargassum zhangii in Palk Strait, southeast India. Hundreds of natural populations of this seaweed were observed in the collection site. BLASTn similarity search using nuclear DNA internal transcribed spacer1 (ITS1) and mitochondrial DNA cytochrome oxidase 1 (COX?1) gene of this isolate indicated Sargassum zhangii as the most homologous sequence available in the repository. Pairwise distances among these isolates were very low, 2.8?נ10?3 and 1.3?נ10?2 with ITS1 and COX?1, respectively, which suggests conspecificity and a recent introduction. Phylogenetic analyses along with other members of genus Sargassum conducted using Bayesian Inference resulted in well-resolved phylograms with robust clades comprising two isolates of S.?zhangii, further confirming conspecificity. With this first report of this seaweed outside China, invasive potential of S.?zhangii is highlighted that warrants immediate global attention. As this is the first time that molecular systematics have been used to catalogue species invasion in India, its efficacy, as well as the necessity for further cataloguing of species invasions using this approach, are highlighted. ? 2016 Dipartimento di Biologia, Universit? di Firenze.Item Strong endemism of bloom-forming tubular Ulva in Indian west coast, with description of Ulva paschima Sp Nov (Ulvales, Chlorophyta)(PLOS One, 2014) Bast, Felix; John, Aijaz Ahmad; Bhushan, SatejUlva intestinalis and Ulva compressa are two bloom-forming morphologically-cryptic species of green seaweeds widely accepted as cosmopolitan in distribution. Previous studies have shown that these are two distinct species that exhibit great morphological plasticity with changing seawater salinity. Here we present a phylogeographic assessment of tubular Ulva that we considered belonging to this complex collected from various marine and estuarine green-tide occurrences in a ca. 600 km stretch of the Indian west coast. Maximum Likelihood and Bayesian Inference phylogenetic reconstructions using ITS nrDNA revealed strong endemism of Indian tubular Ulva, with none of the Indian isolates forming part of the already described phylogenetic clades of either U. compressa or U. intestinalis. Due to the straightforward conclusion that Indian isolates form a robust and distinct phylogenetic clade, a description of a new bloom-forming species, Ulva paschima Bast, is formally proposed. Our phylogenetic reconstructions using Neighbor-Joining method revealed evolutionary affinity of this new species with Ulva flexuosa. This is the first molecular assessment of Ulva from the Indian Subcontinent.Item Strong endemism of bloom-forming tubular ulva in Indian west coast, with description of Ulva paschima sp. nov. (Ulvales, chlorophyta)(Public Library of Science, 2014) Bast, Felix; John, Aijaz Ahmad; Bhushan, SatejUlva intestinalis and Ulva compressa are two bloom-forming morphologically-cryptic species of green seaweeds widely accepted as cosmopolitan in distribution. Previous studies have shown that these are two distinct species that exhibit great morphological plasticity with changing seawater salinity. Here we present a phylogeographic assessment of tubular Ulva that we considered belonging to this complex collected from various marine and estuarine green-tide occurrences in a ca. 600 km stretch of the Indian west coast. Maximum Likelihood and Bayesian Inference phylogenetic reconstructions using ITS nrDNA revealed strong endemism of Indian tubular Ulva, with none of the Indian isolates forming part of the already described phylogenetic clades of either U. compressa or U. intestinalis. Due to the straightforward conclusion that Indian isolates form a robust and distinct phylogenetic clade, a description of a new bloom-forming species, Ulva paschima Bast, is formally proposed. Our phylogenetic reconstructions using Neighbor-Joining method revealed evolutionary affinity of this new species with Ulva flexuosa. This is the first molecular assessment of Ulva from the Indian Subcontinent. ? 2014 Bast et al.