• Issue front cover thumbnail

      Volume 44, Issue 1

      March 2019

    • Delivery of BACE1 siRNA mediated by TARBP-BTP fusion protein reduces β-amyloid deposits in a transgenic mouse model of Alzheimer’s disease

      MOHAMED MOHAMED HAROON KAMAL SABA VENKATA HARSHAVARDHAN BODDEDDA JERALD MAHESH KUMAR ANANT BAHADUR PATEL VIJAYA GOPAL

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      Systemic delivery of nucleic acids to the central nervous system (CNS) is a major challenge for the development of RNAinterference-based therapeutics due to lack of stability, target specificity, non-permeability to the blood–brain barrier (BBB),and lack of suitable carriers. Using a designed bi-functional fusion protein TARBP-BTP in a complex with siRNA, weearlier demonstrated knockdown of target genes in the brain of both AbPP-PS1 (Alzheimer’s disease, AD) and wild-typeC57BL/6 mice. In this report, we further substantiate the approach through an extended use in AbPP-PS1 mice, which upontreatment with seven doses of b-secretase AbPP cleaving Enzyme 1 (BACE1) TARBP-BTP:siRNA, led to target-specificeffect in the mouse brain. Concomitant gene silencing of BACE1, and consequent reduction in plaque load in the cerebralcortex and hippocampus ([60%) in mice treated with TARBP-BTP:siRNA complex, led to improvement in spatial learningand memory. The study validates the efficiency of TARBP-BTP fusion protein as an efficient mediator of RNAi, givingconsiderable scope for future intervention in neurodegenerative disorders through the use of short nucleic acids as genespecific inhibitors.

    • Bone marrow mesenchymal stem cells protect lungs from smoke inhalation injury by differentiating into alveolar epithelial cells via Notch signaling

      YUAN LIANG CUNPING YIN XI LU HUA JIANG FAGUANG JIN

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      To examine the protective effect of transplanting bone marrow mesenchymal stem cells (BMSCs) in treating lung injuryinduced by smoke exposure and to investigate the underlying mechanisms of this protection. SD rats were randomlydivided into four groups: normal group, normal + BMSCGFP group, smoke group, and smoke +BMSCGFP group. Todetect lung injury, we measured arterial blood gas, the wet-to-dry weight ratio, and levels of interleukin-1b, tumor necrosisfactor-a, interleukin-10, and interleukin-13 in bronchoalveolar lavage fluid and lung tissues. We also conductedhistopathology examinations. The protein markers of alveolar epithelial cells were measured to determine the BMSCdifferentiation. The protein levels of Notch1, Jagged-1, and Hes-1 also were detected. In the present study, BMSCtransplantation significantly decreased the wet-dry weight ratio of the lung, reduced the production of inflammatorymediators, and alleviated lung injury simply through differentiating into alveolar type II cells and alveolar type I cells.Western blot analysis confirmed that the protein expression of Notch-1, Jagged-1, and Hes-1 increased significantly aftersystemic BMSC transplantation. No significant difference was observed between the normal group and the normal

      + BMSCGFP group. Our findings indicate that systemic transplantation of BMSCs alleviated lung injury induced bysmoke exposure, which may be associated with BMSCs’ ability to differentiate into alveolar-type cells via the Notchsignaling pathway.

    • D-Tryptophan governs biofilm formation rates and bacterial interaction in P. mendocina and S. aureus

      SAHELI GHOSH ASIFA QURESHI HEMANT J PUROHIT

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      Biofilm genesis by Pseudomonas and Staphylococcus sp is associated with biofouling in natural settings. D-Tryptophan (DTrp)inhibits bacterial biofilms and have been proposed for biofouling control applications. In this study, D-Trp significantlyinhibited Pseudomonas mendocina and Staphylococcus aureus cell attachment (biofilm formation) rates on polystyrene96-well microtiter plates in comparison with L-Tryptophan (L-Trp) and mixtures of D-/L-Tryptophan (D-/L-Trp). Theinhibitory effect was greater on P. mendocina, where the rate of cell adherence was declined to 8.7 X 10E5 cells/h from8.0 X 10E6 cells/h (control) in P. mendocina. In S. aureus it was declined to 4.2 9 107 cells/h from 9.2 X 10E7 cells/h(control) at 1 mM concentration. It hindered the intracellular communication and adherence in both the strains, as confirmedby SEM and real time PCR analysis. Addition of D-Trp to preformed biofilms also caused partial disassembly. Intraand interbacterial aggregation were decreased subsequently upon treatment with D-Trp. It repressed the genes involved incell–cell communication, which could be responsible for the diminished biofilm formation of the selected strains. HenceD-Tryptophan has proved to be an effective strategy to control biofilm and may support in the development of surfacecoating technologies.

    • MusatransSSRDB (a transcriptome derived SSR database) – An advanced tool for banana improvement

      BACKIYARANI SUTHANTHIRAM ARUMUGAM CHANDRASEKAR SUBBARAYA UMA MARIMUTHU SOMASUNDARAM SARASWATHI

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      Availability of transcriptome datasets for use in accelerated molecular-based breeding in Musa species is limited. IlluminaHiseq technology was employed to determine differential gene expression between the contrasting cultivars for threedifferent stresses (Eumusae leaf spot –Mycosphaerella eumusae, root lesion nematode – Pratylenchus coffeae and moisturedeficit stress) under challenged and unchallenged conditions. An average of 34.72 million of reads was assembled into

      *47629 contigs, and *5,466 simple sequence repeats (SSR) from each library were identified. GO annotation and KEGGpathway analysis were carried for all the transcripts and the SSR, SNPs were also detected. Based on this information, aMusatransSSRDB has been developed. Currently, the database consists of 32,800 SSRs with the unique information likeputative function of the SSR-containing genes and their metabolic pathway and expression profiling under various stressconditions. This database provides information on in silico polymorphic SSRs (2830 SSRs) between the contrastingcultivars for each stress and within stress. Information on in silico polymorphic SSRs specific to differentially expressedgenes under challenged condition for each stress can also be accessed. This database facilitates the retrieval of results bynavigating the tabs for cultivars, stress and polymorphism. This database was developed using HTML, Java and PHP;datasets are stored in MySQL database and accessible in the public domain (http://bioinfnrcb.byethost7.com/nrcbbio/). Thisunique information facilitates the banana breeder to select the SSR primers based on specific objectives. MusatransSSRDBalong with other genomics databases will facilitate the genetic dissection and breeding for complex traits in banana. Thus,this database is a step forward in economizing cost, time, manpower and other resources.

    • Carbohydrates and polyphenolics of extracts from genetically altered plant acts as catalysts for in vitro synthesis of silver nanoparticle

      ABHISHEK KUMAR AARAM A KUMAR ADITYA P NAYAK PRIYANKA MISHRA MADHUSMITA PANIGRAHY PRATAP K SAHOO KISHORE C S PANIGRAHI

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      Eco-friendly biosynthetic approach for silver nanoparticles production using plant extracts is an exciting advancement inbio-nanotechnology and has been successfully attempted in nearly 41 plant species. However, an established model plantsystem for systematically unraveling the biochemical components required for silver nanoparticles production is lacking.Here we used Arabidopsis thaliana as the model plant for silver nanoparticles biosynthesis in vitro. Employing biochemical,spectroscopic methods, selected mutants and over-expressor plants of Arabidopsis involved in pleotropicfunctions and sugar homeostasis, we show that carbohydrates, polyphenolics and glyco-proteins are essential componentswhich stimulated silver nanoparticles synthesis. Using molecular genetics as a tool, our data enforces the requirement ofsugar conjugated proteins as essentials for AgNPs synthesis over protein alone. Additionally, a comparative analysis ofAgNPs synthesis using the aqueous extracts of some of the plant species found in a brackish water ecosystem (Gracilaria,Potamogeton, Enteromorpha and Scendesmus) were explored. Plant extract of Potamogeton showed the highest potentialof nanoparticles production comparable to that of Arabidopsis among the species tested. Silver nanoparticles production inthe model plant Arabidopsis not only opens up a possibility of using molecular genetics tool to understand the biochemicalpathways and components in detail for its synthesis.

    • Isolation of a putative virulence agent, cytotoxic serine-elastase, from a newly isolated Pseudomonas aeruginosa ZuhP13

      ESSAM KOTB YEHIA A EL-ZAWAHRY GHADEER E SALEH

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      A 48 kDa ZuhP13 elastase from P. aeruginosa isolated from a urine sample was successfully purified to 8.8-fold and 39%recovery by DEAE-Sepharose CL-6B and Sephadex G-100 chromatography. Its ideal reaction values were pH 7.5 and40 degC. It showed stability at pH 6–9 for 1 h and up to 60 degC for 30 min with midpoint temperature (Tm) at 61.3 degCand isoelectric value (pI) at 5.6±0.2. Its Km and catalytic efficiency (Kcat/Km) for the substrate azocasein were 1.3 mg/mLand 4.62X107 M-1s-1, respectively. On contrary to most P. aeruginosa proteases, Zn2?, EDTA, 2,20-bipyridine ando-phenanthroline showed slight inhibition upon its activity, while, the elastase inhibitors (elastatinal and elastase inhibitorII) and the serine protease inhibitors (TLCK, PMSF, SBTI, and aprotinin) markedly decreased the enzymatic activity. Takentogether, we suggest that ZuhP13 is a serine elastase-type. Interestingly, the tested enzyme showed both hemolytic andhemorrhagic activities in vivo. Furthermore, it induced nuclear lysis yielding hyperchromatism within leaky and malformedhepatocytes, suggesting ZuhP13 elastase as a high molecular weight potential pathological agent.

    • Influence of some environmental variables and addition of r-lysozyme on efficacy of Vibrio harveyi phage for therapy

      TANMOY GON CHOUDHURY BISWAJIT MAITI MN VENUGOPAL INDRANI KARUNASAGAR

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      Bacteriophage therapy is a viable proposition for controlling luminous vibriosis caused by Vibrio harveyi in shrimpaquaculture. However, environmental factors influence the growth and activity of phage and affect its efficiency incontrolling bacterial diseases. An essential problem in the use of vibrio phage as a therapeutic agent was the development ofresistance to phage attachment, rendering them resistant to the lytic action of phage. This problem could be overcome byapplying a cocktail of phages. This study aimed to evaluate the effect of salinity and pH on the phage activity and also tostudy the role of recombinant shrimp lysozyme on the performance of the V. harveyi phage. Out of three different levels ofsalinity (20, 25 and 30 ppt) and pH (6, 7 and 8) tested, optimum phage activity was observed at a salinity of 25 ppt and atneutral pH. Application of recombinant shrimp lysozyme in combination with V. harveyi phage significantly improved theactivity of phage in in vitro assay as well as in microcosm study using seawater. The application of phage along withlysozyme can be a useful approach to overcome the inability of phage to enter the bacteria and thus eliminate or reduce fish/shrimp pathogenic bacteria in aquaculture.

    • Construction of a dietary-cure Saccharomyces cerevisiae expressing long-acting glucagon-like peptide-1 and investigation of its hypoglycemic activity in type 2 diabetes mellitus mouse model

      PINGZHE JIANG CHAO MA JUNFENG HAO YIKE HAN MINGGANG LI

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      Bio-drug is a new type of beneficial biology expressing therapeutic peptides (protein) as orally administrated medicine totreat diseases, in particular, chronic diseases like diabetes. In order to develop recombinant yeast strains as a bio-drug whichcould effectively ameliorate type 2 diabetes, an integrating expression vector pNK1-PGK that could successfully expressgreen fluorescent protein (GFP) in Saccharomyces cerevisiae was constructed to demonstrate the normality of the function.A pNK1-PGK vector, which expresses 10 tandem repeats of long-acting glucagon-like peptide-1(10laGLP-1), was clonedand then transformed into the S. cerevisiae INVSc1. The long-acting GLP-1 hypoglycemic yeast (LHY168) that grewrapidly and expressed 10laGLP-1 stably was screened by uracil-deficient plates and Western blot. The expression quantityof 10laGLP-1 reached 1.56 mg/g cell wet weight. Moreover, the oral administration of LHY168 significantly declined theblood glucose in type 2 diabetic mice that were constructed through co-induction of streptozotocin (STZ) and high-fat andhigh-sugar diet.

    • Measles virus phosphoprotein inhibits apoptosis and enhances clonogenic and migratory properties in HeLa cells

      SANKHAJIT BHATTACHARJEE RISHI KUMAR JAISWAL PRAMOD KUMAR YADAVA

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      Measles virus is the causative agent of measles, a major cause of child mortality in developing countries. Two majorproteins, coded by the viral genome, are nucleocapsid protein (N) and phosphoprotein (P). The N protein protects the viralgenomic RNA and forms ribonucleoprotein complex (RNP) together with P protein. MeV-P protein recruits the largeprotein (L), i.e. viral RNA-depended RNA polymerase (RdRp), to ensure viral replication in host cell. Apoptogenicproperties of N protein of Edmonston vaccine strain have been established in our lab previously. We investigated the role ofMeV-P protein of Edmonston vaccine strain as modulator of apoptosis in cervical cancer cell line (HeLa) and found thatMeV-P protein is anti-apoptotic and enhances cell proliferation. Measles virus is considered to be innately oncotropic virus.However, the anti-apoptotic property of MeV-P protein raises important concerns while adopting this virus as an anti-cancertherapeutic tool.

    • Identification of the miRNA-mRNA regulatory network of antler growth centers

      BAOJIN YAO MEI ZHANG MEIXIN LIU BOCHENG LU XIANGYANG LENG YAOZHONG HU DAQING ZHAO YU ZHAO

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      Antler growth is a unique event compared to other growth and development processes in mammals. Antlers grow extremelyfast during the rapid growth stage when growth rate peaks at 2 cm per day. Antler growth is driven by a specificendochondral ossification process in the growth center that is in the distal region of the antler tip. In this study, we usedstate-of-art RNA-seq technology to analyze the expression profiles of mRNAs and miRNAs during antler growth. Ourresults indicated that the expression levels of multiple genes involved in chondrogenesis and endochondral ossification,including Fn1, Sox9, Col2a1, Acan, Col9a1, Col11a1, Hapln1, Wwp2, Fgfr3, Comp, Sp7 and Ihh, were significantlyincreased at the rapid growth stage. Our results also indicated that there were multiple differentially expressed miRNAsinteracting with differentially expressed genes with opposite expression patterns. Furthermore, some of the miRNAs,including miR-3072-5p, miR-1600, miR-34-5p, miR-6889-5p and miR-6729-5p, simultaneously interacted with andcontrolled multiple genes involved in the process of chondrogenesis and endochondral ossification. Therefore, we establisheda miRNA-mRNA regulatory network by identifying miRNAs and their target genes that were differentially expressedin the antler growth centers by comparing the rapid growth stage and the initial growth stage.

    • G-quadruplex forming region within WT1 promoter is selectively targeted by daunorubicin and mitoxantrone: A possible mechanism for anti-leukemic effect of drugs

      SAEEDEH GHAZAEY ZIDANLOO ABASALT HOSSEINZADEH COLAGAR HOSSEIN AYATOLLAHI ZAHRA BAGHERYAN

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      Wilms tumor 1 (WT1) has long been overexpressed in acute myeloid leukemia and has a prognostic value in its diagnosis.Lately, the formation of G-quadruplexes in oncogenic promoters like WT1 has been widely investigated since stabilizationof these structures leads to transcriptional inhibition of the oncogene. Daunorubicin and mitoxantrone considered as crucialcomponents of almost all standard acute myeloid leukemia induction regimens. Herein we have proposed a probablemolecular mechanism of action through which the drugs may stabilize WT1 promoter G-quadruplexes. Differential pulsevoltammetry, circular dichroism, and polyacrylamide gel electrophoresis, electrophoretic mobility shifts assay, polymerasechain reaction (PCR) stop assays, and quantitative RT-PCR were performed in order to better understanding the nature ofinteractions between the drugs and G-quadruplexes. Data revealed that both drugs had potential to stabilize G-quadruplexesand down-regulate WT1 transcription but daunorubicin exposed more silencing impact. The results illustrated the therapeuticassociation of these two commercial FDA-approved drugs in WT1 transcriptional down-regulation. Since WT1 hasknown as a transcriptional regulator of at least 137 target genes, so the new data are significant for the development of newapproaches to regulating WT1 and other target genes by employing special drugs in cancer treatment.

    • Nest architecture and colony characteristics of Meliponula (Axestotrigona) ferruginea (Hymenoptera, Apidae, Meliponini) in Cameroon

      MOSES TITA MOGHO NJOYA SEINO RICHARD AKWANJOH DIETER WITTMANN

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      Stingless bees have evolved adaptive nest constructions strategies which have resulted in sophisticated nest architecture inmany species while others lack certain structural components. However, no information exists on the nest biology andecology on the genus Meliponula in Cameroon. This study aims to contribute to knowledge on the nest architecture andcolony characteristics of Meliponula (Axestotrigona) ferruginea. Meliponula ferruginea belongs to the genus Meliponulaand subgenus Axestotrigona. This species was first described by Le Peletier De Saini-Farrgeau (1836) and Michener (Thebees of the world, The Johns Hopkins University Press, Baltimore and London, 2000) recently. In Cameroon the specieshas been collected in the northern parts of the country, but there has been no attempt to describe the nest architecture. Ourstingless bee survey from the Bamenda highlands afromontane forests of Cameroon reveals that this species can either nestin tree trunks or in abandoned traditional hollow hives. Interestingly, 50% of colonies studied nested in traditional hollowhives originally baited for honeybees (Apis mellifera adansonii). The nest entrance of Meliponula (Axestotrigona) ferruginearanged from 1 to 1.5 cm (1.2±0.0.24 cm) in diameter, while the external entrance tube extended up to 1.6±0.0.4cm and the nest entrance tube can go up to 5 cm inward in depending on the thickness of the tree trunk. The nest is made upof sticky brown 5–7 (6±0.98) involucra sheets. The size of brood area depends on the age and condition of the nest. Thelength of the nest could be between 5.5 and 7.6 cm (6.8±0.98 cm) and up to 13± 4.16 cm combs with a comb diameter ofabout 7.5±1.29 cm. The cells and storage pots are oval in shape but the storage pots are much larger, about 3 times the sizeof cell. Generally, this study shows a lot of similarities with other previously studied species in terms of the nestcharacteristics and measurements. However, the above characteristics can be very useful in taxonomical, phylogenetic andecological studies of M. ferruginea.

    • AtMBD4: A methylated DNA binding protein negatively regulates a subset of phosphate starvation genes

      ADWAITA PRASAD PARIDA AMRAPALI SHARMA ARUN KUMAR SHARMA

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      DNA methylation is an important epigenetic modification that governs transcriptional regulation. The methylation mark isread by a special class of proteins called methyl-CpG-binding domain proteins. The role of DNA methylation has beenfound in X-chromosome inactivation, genomic imprinting, transposon silencing, and self-incompatibility. Recently,remodeling of global DNA methylation was demonstrated in Arabidopsis during low phosphate availability. The presentstudy reports that AtMBD4 gene of Arabidopsis negatively regulates phosphate starvation. The T-DNA insertion mutation atthe AtMBD4 locus exhibited altered root architecture as compared to wild-type plants. Using microarray hybridization andanalysis, an increased transcript accumulation of 242 genes was observed in the mutant. Many of these genes were relatedto phosphate transporters and transcription factors, involved in phosphate starvation response. Comparison of data ofatmbd4 mutant with publicly available microarray data of phosphate starvation response indicated the role of AtMBD4protein in phosphate starvation response. Further, promoter analysis of up-regulated genes suggested that cis-regulatoryelements like MBS, W-box, and B1BS are more prominent in the promoters of up-regulated genes. Upon performing amethylation-specific PCR, a decreased DNA methylation in the promoter regions of up-regulated genes was observed. Theaccumulation of anthocyanin and inorganic phosphate in the atmbd4 mutant was found to be higher than the wild-typeplant. Altered root morphology, up-regulation of phosphate starvation-induced genes in atmbd4 mutant suggests thatAtMBD4 negatively regulates the phosphate starvation response.

    • Characterization and DNA methylation modulatory activity of gold nanoparticles synthesized by Pseudoalteromonas strain

      YUGANDHARA M PATIL SHRIRAM N RAJPATHAK DEEPTI D DEOBAGKAR

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      Marine extremophiles are shown to tolerate extreme environmental conditions and have high metal reducing properties.Here, we report intracellular synthesis of gold nanoparticles (AuNP) by marine extremophilic bacteria Pseudoalteromonassp. Bac178 which was isolated from the OMZ of Arabian Sea. Preliminary observations suggest that these bacteria usedifferent pathways which may involves the membrane as well as intracellular proteins for the gold salt reduction. Characterizationof the biosynthesised nanoparticles by various techniques such as Scanning electron microscopy (SEM),Transmission electron microscopy (TEM), X-ray diffraction (XRD) and Energy-dispersive X-ray spectroscopy (EDS)confirmed the presence of crystalline gold. These biologically synthesized AuNP were investigated for cytotoxicity andoxidative stress generation in human normal fibroblast and melanoma cells (A375). As AuNP are envisaged to find manyapplications in the medical field, it was of interest to study the effect of AuNP at the epigenetic level. They were found to benon-cytotoxic, non-genotoxic and non-oxidative stress generating over a range of concentrations. Exposure to these AuNPis observed to cause alterations in global DNA methylation as well as in the expression of DNA methyltransferase (DNMT)genes. Since biosynthesized AuNP are being used in various applications and therapies, their epigenetic modulatory activityneeds careful consideration.

    • LIM kinase 1 acts as a profibrotic mediator in permanent atrial fibrillation patients with valvular heart disease

      QINHUA CHEN RYAN C GIMPLE GUANNAN LI JIANZHOU CHEN HAN WU RAN LI JUN XIE BIAO XU

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      Atrial fibrillation (AF) is the most frequently diagnosed cardiac arrhythmia worldwide. Patients with permanent atrialfibrillation are at an increased risk of developing valvular heart disease. Atrial fibrosis occurs in this pathophysiologicalsetting. LIM kinase 1 (LIMK1) is a serine/threonine kinase that regulates microtubule stability and actin polymerization infibroblasts. LIMK1 has been implicated in the pathogenesis of atrial fibrillation. Clinical data and biopsies of the right atrialappendage were collected from 50 patients with valvular heart disease who underwent heart valve replacement surgery.Data from patients with permanent atrial fibrillation (AF) and patients with sinus rhythm (SR) were compared. We foundthat AF patients had upregulated expression of LIMK1 as well as higher fibrosis. Transforming growth factor-b (TGF-b)stimulation induced the differentiation of cardiac fibroblasts into myofibroblasts as well as upregulated expression ofLIMK1. Downregulation of LIMK1 by siRNA inhibited TGF-b induced fibroblast-myofibroblast transition, as evidencedby the downregulation of the expression of several differentiation markers, namely alpha-smooth muscle actin and type Iand III collagen. Our findings revealed that increased LIMK1 protein levels may contribute to atrial fibrosis, and suggestedthat LIMK1 might be involved in AF development by promoting fibrogenesis associated with TGF-b.

    • Autophagy requires Tip20 in Saccharomyces cerevisiae

      LEI CHEN CHUNLING ZHANG YUANCUN LIANG AIXIN LIU HANSONG DONG SHENSHEN ZOU

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      Autophagy is a highly conserved intracellular degradation pathway in eukaryotic cells that responds to environmentalchanges. Genetic analyses have shown that more than 40 autophagy-related genes (ATG) are directly involved in thisprocess in fungi. In addition to Atg proteins, most vesicle transport regulators are also essential for each step of autophagy.The present study showed that one Endoplasmic Reticulum protein in Saccharomyces cerevisiae, Tip20, which controlsGolgi-to-ER retrograde transport, was also required for starvation-induced autophagy under high temperature stress. Intip20 conditional mutant yeast, the transport of Atg8 was impaired during starvation, resulting in multiple Atg8 punctadispersed outside the vacuole that could not be transported to the pre-autophagosomal structure/phagophore assembly site(PAS). Several Atg8 puncta were trapped in ER exit sites (ERES). Moreover, the GFP-Atg8 protease protection assayindicated that Tip20 functions before autophagosome closure. Furthermore, genetic studies showed that Tip20 functionsdownstream of Atg5 and upstream of Atg1, Atg9 and Atg14 in the autophagy pathway. The present data show that Tip20,as a vesicle transport regulator, has novel roles in autophagy.

    • Insights into the evolution of extracellular leucine-rich repeats in metazoans with special reference to Toll-like receptor 4

      DIPANJANA DHAR DEBAYAN DEY SOUMALEE BASU

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      The importance of the widely spread leucine-rich repeat (LRR) motif was studied considering TLRs, the LRR-containingprotein involved in animal immune response. The protein connects intracellular signalling with a chain of molecularinteractions through the presence of LRRs in the ectodomain and TIR in the endodomain. Domain analyses with humanTLR1-9 reported ectodomain with tandem repeats, transmembrane domain and TIR domain. The repeat number variedacross members of TLR and remained characteristic to a particular member. Analysis of gene structure revealed absence ofcodon interruption with TLR3 and TLR4 as exceptions. Extensive study with TLR4 from metazoans confirmed thepresence of 23 LRRs in tandem. Distinct clade formation using coding and amino acid sequence of individual repeatsillustrated independent evolution. Although ectodomain and endodomain exhibited differential selection pressure, withinthe ectodomain, however, the individual repeats displayed positive, negative and neutral selection pressure depending ontheir structural and functional significance.

    • A glucose-insulin-potassium solution improves glucose intake in hypoxic cardiomyocytes by a differential expression of glucose transporters in a metabolic syndrome model

      R CARBÓ E RODRÍGUEZ

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      Among the last consequences of metabolic syndrome are cardiovascular complications such as infarcts. The hypoxic heartswitches its lipid-based metabolism to carbohydrates, and a glucose-insulin-potassium (GIK) solution can be the metabolicsupport to protect the organ. Due to the physiology and cardiac risks associated with the metabolic syndrome, we studied theeffect of GIK solution during hypoxia in a metabolic syndrome model by observing the participation of glucose transporters(GLUTs). The metabolic syndrome characteristics were established by giving a 30% sucrose drinking solution to Wistar ratsfor 24 weeks. The GIK solution’s effect on myocyte glucose uptake during hypoxia and oxygenation was observed using acolorimetric method, and Western blot technique visualized the GLUT participation. Oxygenated control myocytes consumed1.7 ± 0.2 μg of glucose per gram of fresh tissue per hour using the GLUT1, and during hypoxia, they incorporated 41.1%more glucose by GLUT1 and GLUT4. The GIK solution improved glucose uptake in oxygenation by 70.5% through GLUT1.In hypoxia, the uptake was 21% more than the hypoxic control group and by both GLUTs too. Oxygenated metabolicsyndrome myocytes uptake was similar to control cells but achieved by both carriers in oxygenation and hypoxia. Also, theGIK solution had a better response in both oxygenation (113%) and hypoxia (71%). Despite the metabolic energy disorders ofthis syndrome, the GIK solution protects cardiomyocytes, in conditions of hypoxia, through the modulation of both GLUTs.So, this solution can be considered a useful resource during a heart attack in cases of metabolic syndrome.

    • Comparative physiological and leaf proteome analysis between drought-tolerant chickpea Cicer reticulatum and drought-sensitive chickpea C. arietinum

      SERTAN ÇEVIK GÜRLER AKPINAR AYTUNÇ YILDIZLI MURAT KASAP KÜBRA KARAOSMANOĞLU SERPIL ÜNYAYAR

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      Comparative physiological and proteomic analysis were performed to understand the stress responses of two chickpeaspecies (C. reticulatum and C. arietinum) against drought. Our study revealed that drought stress reduced root length, leafwater content, and enhanced free proline content in both species. Effect of drought stress appeared to be greater in C.arietinum compared to C. reticulatum. A total of 24 differently expressed proteins were identified by using MALDI-TOF/TOF-MS/MS in response to drought. The proteins involved in photosynthesis and energy mechanisms were up-regulated inC. reticulatum and down-regulated in C. arietinum under drought. Our results suggest that the photosynthesis capacity of C.reticulatum is greater than that of C. arietinum under drought stress. Abundance of proline and sucrose biosynthesis relatedproteins, glutamine synthetase and cyctosolic fructose-bisphosphate aldolase, respectively, also increased in C. reticulatumunder drought stress. The findings of this proteome analysis will help in understanding the mechanism of drought resistancein chickpea and may be also helpful in developing drought-resistant transgenic plants.

    • Overview of genomics and post-genomics research on type 2 diabetes mellitus: Future perspectives and a framework for further studies

      BATTINI MOHAN REDDY RAYABARAPU PRANAVCHAND S A A LATHEEF

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      In this review, we briefly outlined salient features of pathophysiology and results of the genetic association studies hithertoconducted on type 2 diabetes. Primarily focusing on the current status of genomic research, we briefly discussed the limitedprogress made during the post-genomic era and tried to identify the limitations of the post-genomic research strategies. Wesuggested reanalysis of the existing genomic data through advanced statistical and computational methods and recommendedintegrated genomics-metabolomics approaches for future studies to facilitate understanding of the gene-environmentinteractions in the manifestation of the disease. We also propose a framework for research that may be apt fordetermining the effects of urbanization and changing lifestyles in the manifestation of complex genetic disorders like type 2diabetes in the Indian populations and offset the confounding effects of both genetic and environmental factors in thenatural way.

    • Emerging roles of long non-coding RNAs in cancer

      MANJIMA CHATTERJEE SONALI SENGUPTA

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      Cancer is a physiological condition that has both the endogenous and exogenous influences on its progression. It originatesfrom unusual cell growth, where the cells undergo massive genetic alterations, bypass the signaling machinery andcompromise its genetic cohesion. Literature has well narrated the DNA damage studies including driver mutations thatinterfere with the treatment strategies. However, with evolving medical excellence, recent day studies are trying to unveilthe contribution of RNAs in the progression of tumor malignancies. A number of non-coding RNAs have been identified asan active component in cancer genomics. This article aims to review the role of long non-coding RNAs in the spectra ofcancers and its prognostic value as the biomarkers in molecular targeting with clinical utility and therapeutic beneficence.

    • Mechanistic role of perfusion culture on bone regeneration

      BHASKAR BIRRU NAVEEN KUMAR MEKALA SREENIVASA RAO PARCHA

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      Bone tissue engineering (BTE) aims to develop engineered bone tissue to substitute conventional bone grafts. To achievethis, culturing the cells on the biocompatible three-dimensional (3D) scaffold is one alternative approach. The newfunctional bone tissue regeneration could be feasible by the synergetic combinations of cells, biomaterials and bioreactors.Although the field of biomaterial design/development for BTE applications attained reasonable success, development ofsuitable bioreactor remains still a major challenge. Tissue engineering bioreactors provide the microenvironment requiredfor neo-tissue regeneration, and also can be used to study the physio-chemical cues effect on cell proliferation anddifferentiation in order to produce functional tissue. In this direction, various bioreactors have been developed andevaluated for the successful development of engineered bone tissue. Continues assessment of tissue development andlimitations of the bioreactors lead to the progression of perfusion flow bioreactor system. Improvements in perfusion reactorsystem were able to yield multiple tissue engineered constructs with uniform cell distribution, easy to operate protocols andalso effectively handled for the functional tissue development to meet the adequate supply of engineered graft for clinicalapplication.

    • Interrelation of Ca$^{2+}$ and PE_PGRS proteins during Mycobacterium tuberculosis pathogenesis

      LAXMAN S MEENA

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      In today’s era tuberculosis is a major threat to human population. The lethality of this disease is caused by very efficientlythrived bacteria Mycobacterium tuberculosis (M. tuberculosis). Ca2? plays crucial role in maintenance of cellular homeostasis.Bacilli survival in human alveolar macrophages majorly depends on disruption in Ca2? signaling. Bacilli sustainabilityin phagosome lies in the interruption of phagolysosomal fusion, which is possible because of low intracellularCa2? concentration. Bacilli contain various Ca2? binding proteins which help in regulation of Ca2? signaling for its ownbenefit. For the survival of pathogen, it requires alteration in normal Ca2? concentration in healthy cell. In this review weaim to find the various Ca2? binding domains which are present in several Ca2? binding proteins of M. tuberculosis andvariety of roles played by Ca2? to survive bacilli within host cell. This manuscript emphasizes the Ca2? binding domainspresent in PE_PGRS group of gene family and their functionality in M. tuberculosis survival and pathogenesis.

    • Decoding the biology of language and its implications in language acquisition

      D R RAHUL R JOSEPH PONNIAH

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      Associating human genetic makeup with the faculty of language has long been a goal for biolinguistics. This stimulated theidea that language is attributed to genes and language disabilities are caused by genetic mutations. However, application ofgenetic knowledge on language intervention is still a gap in the existing literature. In an effort to bridge this gap, this articlepresents an account of genetic and neural associations of language and synthesizes the genetic, neural, epigenetic andenvironmental facets involved in language. In addition to describing the association of genes with language, the neural andepigenetic aspects of language are also explored. Further, the environmental aspects of language such as language input,emotion and cognition are also traced back to gene expressions. Therefore, effective language intervention for languagelearning difficulties must offer genetics-informed solutions, both linguistic and medical.

    • Light and auxin signaling cross-talk programme root development in plants

      SONY KUMARI KISHORE C S PANIGRAHI

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      Root development in plants is affected by light and phytohormones. Different ranges of light wavelength influence rootpatterning in a particular manner. Red and white light promote overall root development, whereas blue light has bothpositive as well as negative role in these processes. Light-mediated root development primarily occurs through modulationof synthesis, signaling and transport of the phytohormone auxin. Auxin has been shown to play a critical role in rootdevelopment. It is being well-understood that components of light and auxin signaling cross-talk with each other. However,the signaling network that can modulate the root development is an intense area of research. Currently, limited informationis available about the interaction of these two signaling pathways. This review not only summarizes the current findings onhow different quality and quantity of light affect various aspects of root development but also present the role of auxin inthese developmental aspects starting from lower to higher plants.

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