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Additional file 1: Table S1. Antimalarial Screening of compounds. Figure S1. Molecular structure of all the compounds (n =30) used in this study. Figure S2. Antimalarial effect of galactal fused pyrano-pyranone carbohybrid 12 in chloroquine resistant RKL 9 strain.

Biofilm related bacterial infection is one of the primary causes of implant failure. Limiting bacterial adhesion and colonization of pathogenic bacteria is a challenging task in health care. Here, a highly simplistic processing technique for imparting antibacterial properties on a biomedical grade stainless steel is demonstrated. Low-temperature high strain-rate deformation achieved using submerged friction stir processing resulted in a nearly single phase ultra-fine grain structure. The processed stainless steel demonstrated improved antibacterial properties for both Gram-positive and Gram-negative bacteria, significantly impeding biofilm formation during the in vitro study. Also, the processed stainless steel showed better compatibility with human fibroblasts manifested through apparent cell spreading and proliferation. The substantial antibacterial properties of the processed steel are explained in terms of the favorable electronic characteristics of the metal-oxide and by using classical Derjaguin–Landau–Verwey–Overbeek (DLVO) and the extended DLVO (XDLVO) approach at the cell–substrate interface.

Lignin is a complex organic material connecting cells, fibers, and vessels which constitute wood and other lignified components of the vascular plants. Because of its complex structure consolidated by three-dimensional cross-linking, it is highly inflexible and recalcitrant. The degradation of lignin is required for the efficient utilization of lignocellulosic biomass for the production of biofuels and other cellulose-based products. Removal of lignin is also necessary for the production of paper from wood during the process of delignification and bleaching. Existing physico-chemical methods of lignin degradation are cost and energy incentive and result in generation of toxic waste products. Biodegradation of lignin involving lignin-degrading microbes and their enzyme system is more efficient and environmentally sound approach. Biodegradation of lignin is considered as sustainable technology for the disposal of lignocellulosic biomass and its utilization for the production of value-added products. Among microbes, white rot fungi efficiently degrade lignin with the help of their oxidative extracellular ligninolytic enzymes in general and laccases (benzenediol oxygen oxidoreductase, EC 1.10.3.2) in particular. Laccases are multinuclear enzymes which effectively degrade lignin. In the presence of suitable mediators, laccases hold the potential to oxidize non-phenolic proportion of lignin and other substrates. In the backdrop of aforesaid context, this chapter is an attempt to put forth the details of role of fungal laccases and laccase mediator system in degradation of lignin.

Background: Artesunate the most potent antimalarial is widely used for the treatment of multidrug-resistant malaria. The antimalarial cytotoxicity of Artesunate has been mainly attributed to its selective, irreversible and iron-catalyzed radical mediated damage of parasite biomolecules. In the present research, iron nanoparticle fortified artesunate was tested in P. falciparum and in an experimental malaria mouse model for enhancement in the selectivity and toxicity of Artesunate towards parasite. Artesunate was fortified with nontoxic biocompatible surface modified iron nanoparticle which is specially designed and synthesized for sustained pH-dependent release of Fe2+ within the parasitic food vacuole for enhanced ROS spurt. Methods: Antimalarial efficacy of Iron nanoparticle fortified Artesunate was evaluated in Plasmodium falciparum culture grown in O +ve human blood and in Plasmodium berghei ANKA infected swiss albino mice. Infected mice were daily administered intraperitoneally with Artesunate, nanoparticle fortified Artesunate and monitored for parasitemia. Internalization of iron nanoparticles, pH dependent release of Fe2+, production of reactive oxygen species and parasite biomolecule damage by iron nanoparticle fortified Artesunate was studied using various biochemical, biophysical, ultra-structural and fluorescence microscopy. Results: The nanoparticle fortified Artesunate was highly efficient in 1/8th concentration of Artesunate IC50 and led to retarded growth of P. falciparum with significant damage to macromolecules mediated via enhanced ROS production. Similarly, preclinical In vivo studies also signified a radical reduction in parasitemia with ~8-10-fold reduced dosage of Artesunate when fortified with iron nanoparticles. Interpretation: Surface coated iron-nanoparticle fortified Artesunate can be developed into a potent therapeutic agent towards multidrug-resistant malaria in humans. Funding Statement: This study is supported by Center for Study of Complex malaria in India funded by National Institute of Health, USA, and by Department of Science and Technology (DST-INDIA) and LRE JNU. DK and NY are supported by Shiv Nadar Foundation fellowships. SA is funded by ICMR, Government of India. Declaration of Interests: The authors declare that there are no conflicts of interest. Ethics Approval Statement: Animal studies were performed in accordance with guidelines of the Institutional Animal Ethics Committee (IEAC) of Jawaharlal Nehru University, Delhi and Committee for Control and Supervision of Experiments on Animals (CPCSEA).

Plasmodial resistance to a variety of plant-based antimalarial drugs has led toward the discovery of more effective antimalarial compounds having chemical or biological origin. Since natural compounds are considered as safer drugs, in this study, yeast strains were identified and compared for the production of carotenoids that are well-known antioxidants and this metabolite was tested for its antiparasitic activity. Plasmodium falciparum 3D7 strain was selected as the target parasite for evaluation of antimalarial activity of yeast carotenoids using in vitro studies. Data were analyzed by FACS (fluorescence-activated cell sorter) and counted via gold standard Giemsa-stained smears. The extracted yeast carotenoids showed a profound inhibitory effect at a concentration of 10–3 µg/µl and 10−4 µg/µl when compared to β- carotene as control. SYBR Green1 fluorescent dye was used to confirm the decrease in parasitaemia at given range of concentration. Egress assay results suggested that treated parasite remained stalled at schizont stage with constricted morphology and were darkly stained. Non-toxicity of carotenoids on erythrocytes and on human liver hepatocellular carcinoma cells (HepG2 cells) was shown at a given concentration. This report provides strong evidence for antimalarial effects of extracted yeast carotenoids, which can be produced via a sustainable and cost-effective strategy and may be scaled up for industrial application.

A successful root canal treatment depend upon so many factor but the most important factor which determine the efficacy of all subsequent procedures such as debridement ,medicament delivery, and obturation is mechanical preparation of root canal system .Traditionally stainless steel files were used, but with the introduction of nickel titanium (NiTi) alloys significantly broadened the instrument design. So many factors affect the separation of nickel titanium instrument and the rate of separation of nickel titanium is 30-60% and breakage rate 9.4%. The aim of this review article is to discuss the methods that prevent the separation of nickel titanium instrument and improve the success of root canal treatment to save teeth.

Bioisosterism is a useful strategy in rational drug design to improve pharmacodynamic and pharmacokinetic properties of lead compounds. Imidazolidinones have been reported as potent kinase inhibitors and antileishmanial agents. In this study, bioisosteres of imidazolidinones (compounds 1-3) were evaluated for their antileishmanial properties. The modified imidazolidinones exhibited potent antileishmanial activity against extracellular as well as intracellular Leishmania donovani parasites in nanomolar concentrations. The selectivity index of these compounds on host cells was found to be more than 1000, emphasizing their specificity toward the parasite. Using SwissTargetPrediction software, we assessed the potential targets of these compounds and found MAPK as the most probable target. To in vitro validate, we developed a novel in vitro kinase assay that mimics the in vivo nature of the functional kinome. Compounds 1-3 displayed specific inhibition of parasite kinase activity accompanied by an increase in intracellular sodium levels in the parasites. This might be the effect of kinase inhibition that regulates sodium homeostasis through Na-ATPases. Finally, the compound-treated parasites underwent apoptosis-like death. This study represents bioisoterism as a novel approach for drug design to establish the structure-activity relationship, which in turn helps to improve the therapeutic activity of lead compounds.

Petroleum refinery effluents (PRE) are generally the wastes generated from industries primarily engaged in refining crude oil, manufacturing fuels, lubricants and petrochemical intermediates. These effluents or wastewater, generated, are considered as a major source of aquatic environmental pollution. The effluents are mainly composed of oil, grease and many other toxic organic compounds. The process of crude oil refining consumes large volume of water. Consequently, significant volume of wastewater is generated. The requirement of water depends upon on the size, crude products and complexity of operation. Petroleum refining units need water for distillation, desalting, thermal cracking, catalytic and treatment processes in order to produce useful products such as LPG (Liquefied Petroleum Gas), gasoline, asphalt, diesel, jet fuel, petroleum feedstock etc. Wastewater generated through petroleum refineries contains various hydrocarbons. It has been estimated that the demand for world oil is expected to rise to 107 mbpd (million barrels per day) in the next two decades. By 2030 oil will account for 32% of the world’s energy supply. The increasing demand of oil clearly shows that effluents produced from the oil industry will continue to be produced and discharged into the water bodies. The pollutants found in the effluent are seriously toxic and hazardous to the environment. Techniques used for effluent treatment include adsorption, coagulation, chemical oxidation, biological techniques as well as contemporary technologies like membranes and microwave-assisted catalytic wet air oxidation and Advanced oxidation processes (AOP) like heterogeneous photo-catalytic degradation which is based on its potential to completely mineralize the organic effluents beside being cost effective, readily available and the catalyst used itself is non-toxic in nature. The review provides a detailed description on nature of effluent or wastewater produced from the oil refinery units, its discharge into the water bodies, toxicological effects of the effluent on terrestrial and aquatic ecosystem and the various treatment technologies designed for the treatment and recycling of wastewater generated during operation.

Currently, there is no efficient vaccine available against clinical malaria. However, continuous efforts have been committed to develop powerful antimalarial vaccine by discovery of novel antigens with in-depth understanding of its nature, immunogenicity, and presentation (delivery adjuvants). Moreover, another important part of vaccine development includes discovery of better immunostimulatory formulation components (immunostimulants). A protective vaccine against malaria requires antigen-specific B and T helper cell responses as well as cytotoxic T lymphocyte (CTL) responses. A long-lasting B and T memory cell production is also required for effective malaria vaccine. Since activation of Toll-like receptors (TLRs) promotes both innate inflammatory responses as well as the induction of adaptive immunity, several initiatives have been mounted during the last few years for the use of TLR agonists as malaria vaccine adjuvants.The review summarizes reports related to the use and development of TLR agonists as malaria vaccine adjuvants and describes various strategies involved for the selection of specific antigens and TLR agonists.TLR agonists are promising adjuvants for the development of effective malaria vaccine, allowing for both innate inflammatory responses as well as the induction of adaptive immunity.

Palmitoylation has been recently identified as an important post-translational rheostat for controlling protein function in eukaryotes. However, the molecular machinery underlying palmitoylation remains unclear in the neglected tropical parasite, Leishmania donovani. Herein, we have identified a catalog of 20 novel palmitoyl acyltransferases (PATs) and characterized the promastigote-specific PAT (LdPAT4) containing the canonical Asp-His-His-Cys (DHHC) domain. Immunofluorescence analysis using in-house generated LdPAT4-specific antibody demonstrated distinct expression of LdPAT4 in the flagellar pocket of promastigotes. Using metabolic labeling-coupled click chemistry method, the functionality of this recombinant enzyme could be authenticated in E. coli strain expressing LdPAT4-DHHC domain. This was evident by the cellular uptake of palmitic acid analogs, which could be successfully inhibited by 2-BMP, a PAT-specific inhibitor. Using CSS-Palm based in-silico proteomic analysis, we could predict up to 23 palmitoylated sites per protein in the promastigotes, and further identify distinctive palmitoylated protein clusters involved in microtubule assembly, flagella motility and vesicular trafficking. To highlight, proteins such as Flagellar Member proteins (FLAM1, FLAM5), Intraflagellar Transport proteins (IFT88), and flagellar motor assembly proteins including the Dynein family were found to be enriched. Furthermore, analysis of global palmitoylation in promastigotes using Acyl-biotin exchange purification identified a set of S-palmitoylated proteins overlapping with the in-silico proteomics data. The attenuation of palmitoylation using 2-BMP demonstrated several phenotypic alterations in the promastigotes including distorted morphology, reduced motility (flagellar loss or slow flagellar beating), and inefficient invasion of promastigotes to host macrophages. These analyses confirm the essential role of palmitoylation in promastigotes. In summary, the findings suggest that LdPAT4 acts as a functional acyltransferase that can regulate palmitoylation of proteins involved in parasite motility and invasion, thus, can serve as a potential target for designing chemotherapeutics in Visceral Leishmaniasis.

Microglial cells form a context-dependent network of brain immunoeffector cells. Despite their indispensable roles, unresolved questions exist around biomarker discovery relevant to their cellular localization, self-renewing potential, and brain developmental dynamics. To resolve the existent gap in the annotation of candidate biomarkers, we conducted a meta-analysis of brain cells using available high-throughput data sets for deciphering microglia-specific expression profiles. We have identified 3,290 significant genes specific to microglia and further selected the top 20 dysregulated genes on the basis of p-value and log2FC. To this list, we added 7 known microglia-specific markers making the candidate list comprising 27 genes for further downstream analyses. Next, we established a connectome of these potential markers with their putative protein partners, which demonstrated strong associations of upregulated genes like Dedicator of cytokinesis 2 (DOCK2) with early/mature microglial markers such as Sphingosine kinase 1 (SPHK1), CD68, and CD45. To elucidate their respective brain anatomical location, we deconvoluted the BrainSpan Atlas expression data. This analysis showed high expression of the majority of candidate genes in microglia-dense regions (Amygdala, Hippocampus, Striatum) in the postnatal brain. Furthermore, to decipher their localized expression across brain ages, we constructed a developmental dynamics map (DDM) comprising extensive gene expression profiles throughout prenatal to postnatal stages, which resulted in the discovery of novel microglia-specific gene signatures. One of the interesting readout from DDM is that all the microglia-dense regions exhibit dynamic regulation of few genes at 37 post conception week (pcw), the transition period between pre- and postnatal stages. To validate these findings and correlate them as potential biomarkers, we analyzed the expression of corresponding proteins in hESC-derived human microglia precursors. The cultured microglial precursors showed expression of Pentraxin 3 (PTX3) and SPHK1 as well as several known markers like CD68, Allograft inflammatory factor 1 (AIF1/IBA1). In summary, this study has furnished critical insights into microglia dynamics across human brain ages and cataloged potential transcriptomic fingerprints that can be further exploited for designing novel neurotherapeutics.

Natural products from medicinal plants have always attracted a lot of attention due to their diverse and interesting therapeutic properties. We have employed the principles of green chemistry involving isomerization, coupling and condensation reaction to synthesize a class of compounds derived from eugenol, a naturally occurring bioactive phytophenol. The compounds were characterized structurally by 1H-, 13C-NMR, FT-IR spectroscopy and mass spectrometry analysis. The purity of compounds was detected by HPLC. The synthesized compounds exhibited anti-cancer activity. A 10–12-fold enhancement in efficiency of drug molecules (~ 1 µM) was observed when delivered with graphene oxide (GO) as a nanovehicle. Our data suggest cell death via apoptosis in a dose-dependent manner due to increase in calcium levels in specific cancer cell lines. Interestingly, the benzoxazine derivatives of eugenol with GO nanoparticle exhibited enhanced therapeutic potential in cancer cells. In addition to anti-cancer effect, we also observed significant role of these derivatives on parasite suggesting its multi-pharmacological capability.

This study was carried out to compare the different techniques of placement of polyethylene fibre (Ribbond) on reinforcement of endodontically treated teeth with MOD cavities in vitro. Forty extracted human premolars were randomly assigned to four groups (n = 10). Teeth in Groups I-IV received root canal treatment and a MOD cavity preparation, with gingival cavosurface margin 1.5 mm in coronal to cementoenamel junction. Group I served as no fibre group, Group II as occlusal fibre group, Group III as base fibre group and Group IV as dual-fibre group (occlusal and base both). Subsequent to restoring with composite resin and thermocycling, a vertical compressive force was applied at a cross-head speed of 0.5 mm min(-1) using universal testing machine until fracture. Data were analysed using one-way analysis of variance and Tukey's post hoc tests. Fracture resistance was significantly highest in dual-fibre group (P

Herein we have reported design, synthesis and in vitro biological evaluation of a library of bicyclic lactams that led to the discovery of compounds 6 and 7 as a novel class of α-glucosidase inhibitors. They inhibited α-glucosidase (yeast origin) in a mixed type of inhibition with an IC50 of ∼150 nM. Molecular docking studies further substantiated screening results. Interestingly phenotypic screening of this library against the human malaria parasite revealed 7 as a potent antiplasmodial agent.

Expression of synthetic proteins from intergenic regions of E. coli and their functional association was recently demonstrated (Dhar et al. in J Biol Eng 3:2, 2009. doi: 10.1186/1754-1611-3-2 ). This gave birth to the question: if one can make ‘user-defined’ genes from non-coding genome—how big is the artificially translatable genome? (Dinger et al. in PLoS Comput Biol 4, 2008; Frith et al. in RNA Biol 3(1):40–48, 2006a; Frith et al. in PLoS Genet 2(4):e52, 2006b). To answer this question, we performed a bioinformatics study of all reported E. coli intergenic sequences, in search of novel peptides and proteins, unexpressed by nature. Overall, 2500 E. coli intergenic sequences were computationally translated into ‘protein sequence equivalents’ and matched against all known proteins. Sequences that did not show any resemblance were used for building a comprehensive profile in terms of their structure, function, localization, interactions, stability so on. A total of 362 protein sequences showed evidence of stable tertiary conformations encoded by the intergenic sequences of E. coli genome. Experimental studies are underway to confirm some of the key predictions. This study points to a vast untapped repository of functional molecules lying undiscovered in the non-expressed genome of various organisms.

Malaria a global pandemic has engulfed nearly 0.63 million people globally. It is high time that a cure for malaria is required to stop its ever increasing menace. Our commentary discusses the advent and contribution of diversity oriented synthesis (DOS) in the drug discovery efforts towards developing cure for malaria. DOS based on chemical genetics focusses on design and synthesis of molecular libraries which covers large tracts of biologically relevant chemical space. Herein we will discuss the applications, advantages, disadvantages and future directions of DOS with respect to malaria.

Here we described a natural product inspired modular DOS strategy for the synthesis of a library of hybrid systems that are structurally and stereochemically disparate. The main scaffold is a pyrroloisoquinoline motif, that is synthesized from tandem Pictet-Spengler lactamization. The structural diversity is generated via “privileged scaffolds” that are attached at the appropriate site of the motif. Screening of the library compounds for their antiplasmodial activity against chloroquine sensitive 3D7 cells indicated few compounds with moderate activity (20–50 μM). A systematic comparison of structural intricacy between the library members and a natural product dataset obtained from ZINC ® revealed comparable complexity.

Serpentine receptors with G-protein coupled receptor like seven transmembrane (7 TM) topology are identified in Plasmodium. A class of 7 TM receptors known as purinergic receptors binds to purines such as ADP, ATP and UTP and mediates important physiological functions including regulation of calcium signaling. Here we performed in silico analysis of Plasmodium falciparum serpentine receptors and found that one of the P. falciparum serpentine receptors, PfSR12 possess nucleotide binding consensus P-loop sequence in addition to seven transmembrane domains. The presence of conserved seven transmembrane domains and a consensus nucleotide binding sequence (P-loop) suggest that PfSR12 is a putative purinergic receptor. On further analysis using docking programmes we found four active binding residues Asn149, Lys150, Asn151 and Gly152 in P-loop of PfSR12, interact with ATP. This work gives insights into the interactions between putative purinergic receptor PfSR12 and its ligand ATP which can be explored in structure based drug designing against malaria.

Malaria, a leading parasitic killer, is caused by Plasmodium spp. The pathology of the disease starts when Plasmodium merozoites infect erythrocytes to form rings, that matures through a large trophozoite form and develop into schizonts containing multiple merozoites. The number of intra-erythrocytic merozoites is a key-determining factor for multiplication rate of the parasite. Counting of intraerythrocytic merozoites by classical 2-D microscopy method is error prone due to insufficient representation of merozoite in one optical plane of a schizont. Here, we report an alternative 3-D microscopy based automated method for counting of intraerythrocytic merozoites in entire volume of schizont. This method offers a considerable amount of advantages in terms of both, ease and accuracy.

Peptides are increasingly used as inhibitors of various disease specific targets. Several naturally occurring and synthetically developed peptides are undergoing clinical trials. Our work explores the possibility of reusing the non-expressing DNA sequences to predict potential drug-target specific peptides. Recently, we experimentally demonstrated the artificial synthesis of novel proteins from non-coding regions of Escherichia coli genome. In this study, a library of synthetic peptides (Synpeps) was constructed from 2500 intergenic E. coli sequences and screened against Beta-secretase 1 protein, a known drug target for Alzheimer’s disease (AD). Secondary and tertiary protein structure predictions followed by protein–protein docking studies were performed to identify the most promising enzyme inhibitors. Interacting residues and favorable binding poses of lead peptide inhibitors were studied. Though initial results are encouraging, experimental validation is required in future to develop efficient target specific inhibitors against AD.

The human malaria parasite Plasmodium falciparum possesses sophisticated systems of protein secretion to modulate host cell invasion and remodeling. In the present study, we provide insights into the function of the AP-1 complex in P. falciparum. We utilized GFP fusion constructs for live cell imaging, as well as fixed parasites in immunofluorescence analysis, to study adaptor protein mu1 (Pfμ1) mediated protein trafficking in P. falciparum. In trophozoites Pfμ1 showed similar dynamic localization to that of several Golgi/ER markers, indicating Golgi/ER localization. Treatment of transgenic parasites with Brefeldin A altered the localization of Golgi-associated Pfμ1, supporting the localization studies. Co-localization studies showed considerable overlap of Pfμ1 with the resident rhoptry proteins, rhoptry associated protein 1 (RAP1) and Cytoadherence linked asexual gene 3.1 (Clag3.1) in schizont stage. Immunoprecipitation experiments with Pfμ1 and PfRAP1 revealed an interaction, which may be mediated through an intermediate transmembrane cargo receptor. A specific role for Pfμ1 in trafficking was suggested by treatment with AlF4, which resulted in a shift to a predominantly ER-associated compartment and consequent decrease in co-localization with the Golgi marker GRASP. Together, these results suggest a role for the AP-1 complex in rhoptry protein trafficking in P. falciparum.

Aims: Management of vertically fractured mandibular first molar by intra coronal splinting applying the finite centre of rotation effect of Pythagorus theorem. Case Presentations: A 28 year old male patient presented at the Department of Conservative Dentistry and Endodontics with the chief complaint of pain in the lower right back region of the mouth since last the 10 days. Past dental history revealed that affected tooth had root canal treatment done two years ago. History of present illness was that pain was elicited on chewing, and applying pressure on that area, with occasional bleeding from the same region. Intraoral examination revealed that tooth 46 was tender on percussion with vertically fractured crown in Case Study Tripathi et al.; BJMMR, 7(7): 623-629, 2015; Article no.BJMMR.2015.369 624 mesiodistal direction. Periapical Radiograph revealed that tooth was endodontically treated, with radiolucencies at apical and furcal area, and visible fracture line in mesiodistal direction at CEJ level. Technique Used in the Study: The tooth was endodontically retreated and Pythagorus theorem was used to locate the centres of rotation on both buccal and lingual sides of tooth 46 coronally. The technique was followed by drilling a vent at the same position approximating a fibre post (3M, Relyxfibre post). The centres were different both for buccal and lingual aspects which when approximated will provide antirotation. The post was placed in the buccolingual direction and the tooth was restored with posterior composites after which the restoration was checked for any high points. Follow up of the case was done for the period of one year. Discussion: The concept of the estimation of the finite centre of rotation is like the primary objective in tightening a screw joint to generate an optimum preload that will maximize the fatigue life of the screw while offering a reasonable degree of protection against loosening. The major advantage of this approach is that the fragment was stabilized properly and there was healthy development of periodontium over the year. Conclusion: Usually the vertically fractured tooth have poor prognosis and may likely be referred for extraction if not stabilized properly. This case report emphasizes the basic concept of stabilization and preventing rotation around an axis which will give the fractured tooth a better longevity and avoid the vibrations during crown preparation. The combined effect will restore the patient’s own natural tooth back and minimizes the chances of periodontal complication.

Bilateral inverted impacted third molar is a very rare incidence. Dearth of dental arch length and space are also considered as the chief cause for tooth impaction. Reviewing the literatures, mandibular third molar was the most frequently impacted tooth, followed by the maxillary third molars, the maxillary canines and the mandibular premolars.

Several natural product inspired fused and spiro-heterocyclic scaffolds were prepared by ring distortion and ring construction strategies and evaluated for anti-breast cancer activity. A facile domino Pictet–Spengler lactamization (PSL) afforded nine natural product inspired indolo[2,3-a]quinolizidine and indolo[8,7-b]indolizidine scaffolds which are converted to seven other scaffolds by functional group transformation, ring distortion and ring construction strategies. In vitro screenings of this library of sixteen scaffolds with six distinct architectures against MCF7 cell lines afforded two compounds (10 and 21) with modest activity. Principal component analysis of this library against databases of FDA approved drugs, commercial compounds and FDA approved breast cancer compounds indicated an eclectic mix of structures among the molecules.

The interaction of (7-chloroquinolin-4-yl)-(2,5-dimethoxyphenyl)-amine hydrochloride dihydrate (CQDPA), an amodiaquine analog and a combination partner in antimalarial therapy, with serum albumin proteins (BSA and HSA) was assessed using spectroscopic techniques. Fluorescence studies at three different temperatures confirmed the binding of CQDPA to the active sites of the proteins. The thermodynamic properties such as the enthalpy change (ΔH0), Gibbs free energy change (ΔG0) and entropy change (ΔS0) suggest that the CQDPA molecule binds to site I (subdomain II) of both BSA and HSA through hydrophobic interactions. Based on Forster’s theory of non-radiation energy transfer, the average binding distance r values between the donor (BSA or HSA) and acceptor (CQDPA) were found to be 2.63 and 2.77 for BSA and HSA respectively. A CD study revealed that the α-helical content remained intact in both BSA and HSA on the addition of the amodiaquine analogue but with decreased intensity. The computational analysis and molecular docking of CQDPA with BSA & HSA also corroborated the experimental results.

Kati Basti included under various external procedures of Ayurveda, having variety of actions like the Bahya Snehana (external oleation), Swedana Chikitsa (fomentation therapy). Kati Basti is indicated in various disorders of spine and back like backache, lumbar spondylosis, sciatica, degenerative disc changes etc. Low back pain is most common complaint with a prevalence of 65 to 90%. Improper sitting postures, traveling, use of two wheeler and sports activities are few important causes of backache. Sciatica often used to describe low back pain that spreads (radiates) through the hip, to the back of the thigh, and down the inside of the leg which closely resembles with Gridhrasi. In Sharanghadhara Samhita use of Prabhanjana Taila in Ghridhrasi Vyadhi has been indicated, hence an attempt was made to compare clinically the efficacy of Kati Basti with Prabhanjanam Taila and Moorchita Tila Taila in Gridhrasi with special reference to sciatica.

The human malaria parasite Plasmodium falciparum proliferates in red blood cells following repeated cycles of invasion, multiplication, and egress. P. falciparum serine repeat antigen 5 (PfSERA5), a putative serine protease, plays an important role in merozoite egress. However, regulation of its activity leading to merozoite egress is poorly understood. In this study, we show that PfSERA5 undergoes phosphorylation prior to merozoite egress. Immunoprecipitation of parasite lysates using anti-PfSERA5 serum followed by MS analysis identified calcium-dependent protein kinase 1 (PfCDPK1) as an interacting kinase. Association of PfSERA5 with PfCDPK1 was corroborated by co-sedimentation, co-immunoprecipitation, and co-immunolocalization analyses. Interestingly, PfCDPK1 phosphorylated PfSERA5 in vitro in the presence of Ca(2+) and enhanced its proteolytic activity. A PfCDPK1 inhibitor, purfalcamine, blocked the phosphorylation and activation of PfSERA5 both in vitroas well as in schizonts, which, in turn, blocked merozoite egress. Together, these results suggest that phosphorylation of PfSERA5 by PfCDPK1 following a rise in cytosolic Ca(2+) levels activates its proteolytic activity to trigger merozoite egress.

Development of new class of anti-malarial drugs is an essential requirement for the elimination of malaria. Bioactive components present in medicinal plants and their chemically modified derivatives could be a way forward towards the discovery of effective anti-malarial drugs. Herein, we describe a new class of compounds, 1,3-benzoxazine derivatives of pharmacologically active phytophenols eugenol (compound 3) and isoeugenol (compound 4) synthesised on the principles of green chemistry, as anti-malarials. Compound 4, showed highest anti-malarial activity with no cytotoxicity towards mammalian cells. Compound 4 induced alterations in the intracellular Na+ levels and mitochondrial depolarisation in intraerythrocytic Plasmodium falciparum leading to cell death. Knowing P-type cation ATPase PfATP4 is a regulator for sodium homeostasis, binding of compound 3, compound 4 and eugenol to PfATP4 was analysed by molecular docking studies. Compounds showed binding to the catalytic pocket of PfATP4, however compound 4 showed stronger binding due to the presence of propylene functionality, which corroborates its higher anti-malarial activity. Furthermore, anti-malarial half maximal effective concentration of compound 4 was reduced to 490 nM from 17.54 µM with nanomaterial graphene oxide. Altogether, this study presents anti-plasmodial potential of benzoxazine derivatives of phytophenols and establishes disruption of parasite sodium homeostasis as their mechanism of action.

Substrate-cell interactions for a bioimplant are driven by substrate's surface characteristics. In addition, the performance of an implant and resistance to degradation are primarily governed by its surface properties. A bioimplant typically degrades by wear and corrosion in the physiological environment, resulting in metallosis. Surface engineering strategies for limiting degradation of implants and enhancing their performance may reduce or eliminate the need for implant removal surgeries and the associated cost. In the current study, we tailored the surface properties of stainless steel using submerged friction stir processing (FSP), a severe plastic deformation technique. FSP resulted in significant microstructural refinement from 22 μm grain size for the as-received alloy to 0.8 μm grain size for the processed sample with increase in hardness by nearly 1.5 times. The wear and corrosion behavior of the processed alloy was evaluated in simulated body fluid. The processed sample demonstrated remarkable improvement in both wear and corrosion resistance, which is explained by surface strengthening and formation of a highly stable passive layer. The methylthiazol tetrazolium assay demonstrated that the processed sample is better in supporting cell attachment, proliferation with minimal toxicity, and hemolysis. The athrombogenic characteristic of the as-received and processed samples was evaluated by fibrinogen adsorption and platelet adhesion via the enzyme-linked immunosorbent assay and lactate dehydrogenase assay, respectively. The processed sample showed less platelet and fibrinogen adhesion compared with the as-received alloy, signifying its high thromboresistance. The current study suggests friction stir processing to be a versatile toolbox for enhancing the performance and reliability of currently used bioimplant materials.

During the blood stage, Plasmodium spp. merozoites invade host red blood cells (RBCs), multiply, exit, and reinvade uninfected RBCs in a continuing cycle that is responsible for all the clinical symptoms associated with malaria. Entry into (invasion) and exit from (egress) RBCs are highly regulated processes that are mediated by an array of parasite proteins with specific functional roles. Many of these parasite proteins are stored in specialized apical secretory vesicles, and their timely release is critical for successful invasion and egress. For example, the discharge of parasite protein ligands to the apical surface of merozoites is required for interaction with host receptors to mediate invasion, and the timely discharge of proteases and pore-forming proteins helps in permeabilization and dismantling of limiting membranes during egress. This review focuses on our understanding of the signaling mechanisms that regulate apical organelle secretion during host cell invasion and egress by malaria parasites. The review also explores how understanding key signaling mechanisms in the parasite can open opportunities to develop novel strategies to target Plasmodium parasites and eliminate malaria.

Aim: The present study attempted to find out the relationship between positive and negative clinical symptoms and Wisconsin Card Sorting Test (WCST) performance in a group of schizophrenia patients. Methodology: Fifty schizophrenia patients were assessed using the Positive and Negative Syndrome Scale (PANSS) by a trained psychiatrist (TKA) and two groups, each of 25 positive symptom and 25 negative symptom schizophrenia patients were formed. On these fifty patients with schizophrenia and 15 normal control groups, WCST measures were applied by a clinical psychologist (SS) who remained blind to the PANSS score. Results: Schizophrenia diagnosis significantly affects WCST performances. One-way analysis of variance (ANOVA) revealed schizophrenia patients showed a significant impairment on all WCST indices compared with normal subjects except versus total number of correct responses. Post hoc comparison (Tukey HSD Test) between means revealed that negative schizophrenia patients showed significantly worse performance on most WCST performance parameters: percent errors, perseverative responses, percent perseverative responses, perseverative errors, percent perseverative errors, and conceptual level responses. Conclusions: Both positive and negative symptom schizophrenia patients have some distinct WCST measures deficits.

Adult neurogenesis is primarily directed by neural progenitor cells, which reside in the subventricular zone (SVZ) and subgranular zone (SGZ) of the brain. Unfolding transcriptional heterogeneity and complexity of various neurodevelopmental stages can probe new insights into neurogenesis and neurodevelopmental disorders. Recent findings have suggested that epigenetic regulatory mechanisms in neural differentiation involve long noncoding RNAs (lncRNAs) as a new genre of regulators. Although many studies have addressed the overall consequences of the noncoding RNome (noncoding RNA content) on the genome, lesser is known about their specific roles and consequences in adult neurogenesis, neurodevelopmental stages, and onset of neuropathology. Recent advances in induced pluripotent stem cell (iPSC)-based neurological disease modeling have shed light on new avenues to investigate neuronal development as well as molecular paradigms underlying onset of neurological impairments. However, due to limited availability of brain tissues and gap in the understanding of lncRNA biomarkers in neurodevelopment, the study of lncRNA in neurogenesis still exists at its infancy. To further understand the lncRNA-mediated regulation in stage-specific development of pluripotent stem cell-derived neurons and other brain cells, we identified potential lncRNA signatures implicative in brain development or dysregulation using data mining and analyses. They may be used in monitoring disease progression and may serve as potential targets for novel therapeutic approaches.

Malaria is a major global health problem, caused by Plasmodium sps. Clinical symptoms of the disease are associated with asexual stages of parasite life cycle in human erythrocytes whereas transmission of disease is attributed to sexual stages in mosquito. Increasing resistance to known antimalarial drugs has resulted in the evolution of newer approaches of drug discovery against the disease. The conventional phenotypic screening approaches of drug discovery enable high-throughput screening of a library of chemical compounds for antimalarial effect followed by target identification and its experimental validation. Alternatively, target-based approach for drug discovery involves screening of compounds against known parasite targets by in vitro and in vivo studies. This chapter focuses on the contemporary approaches in antimalarial drug discovery that promise the development of an effective strategy to combat malaria.

Histone lysine methyltransferase (HKMT) are histone-modifying enzymes that catalyze the transfer of methyl groups to lysine and arginine residues of histone protein. HKMTs have been involved in transcriptional regulation of various proteins in organisms. Malaria parasite also has HKMT, which plays a major role in parasite development and pathogenesis and also in regulation of various biological process and pathways. Our aim is to study fundamental biology of key molecules involved in the survival of Plasmodium falciparum and use these to develop efficient synthetic peptides and chemical compounds. As a first step in this direction, we computationally predicted the three-dimensional structure of HKMT of P. falciparum (PfHKMT) by using iterative threading assembly refinement. The PfHKMT three-dimensional model was validated using PROCHECK and docked with known HKMT inhibitor Bix01294 using Autodock. Our initial results are encouraging and indicate that structural analysis of PfHKMT could be important in developing novel synthetic molecules against malaria.

Egress of malaria parasites from the host cell requires the concerted rupture of its enveloping membranes. Hence, we investigated the role of the plasmodial perforin-like protein PPLP2 in the egress of Plasmodium falciparum from erythrocytes. PPLP2 is expressed in blood stage schizonts and mature gametocytes. The protein localizes in vesicular structures, which in activated gametocytes discharge PPLP2 in a calcium-dependent manner. PPLP2 comprises a MACPF domain and recombinant PPLP2 has haemolytic activities towards erythrocytes. PPLP2-deficient [PPLP2(-)] merozoites show normal egress dynamics during the erythrocytic replication cycle, but activated PPLP2(-) gametocytes were unable to leave erythrocytes and stayed trapped within these cells. While the parasitophorous vacuole membrane ruptured normally, the activated PPLP2(-) gametocytes were unable to permeabilize the erythrocyte membrane and to release the erythrocyte cytoplasm. In consequence, transmission of PPLP2( -) parasites to the Anopheles vector was reduced. Pore-forming equinatoxin II rescued both PPLP2(-) gametocyte exflagellation and parasite transmission. The pore sealant Tetronic 90R4, on the other hand, caused trapping of activated wild-type gametocytes within the enveloping erythrocytes, thus mimicking the PPLP2(-) loss-of-function phenotype. We propose that the haemolytic activity of PPLP2 is essential for gametocyte egress due to permeabilization of the erythrocyte membrane and depletion of the erythrocyte cytoplasm.

Merozoite surface protein 3 of Plasmodium falciparum, a 40-kDa protein that also binds heme, has been biophysically characterized for its tendency to form highly elongated oligomers. This study aims to systematically analyze the regions in MSP3 sequence involved in oligomerization and correlate its aggregation tendency with its high affinity for binding with heme. Through size exclusion chromatography, dynamic light scattering, and transmission electron microscopy, we have found that MSP3, previously known to form elongated oligomers, actually forms self-assembled filamentous structures that possess amyloid-like characteristics. By expressing different regions of MSP3, we observed that the previously described leucine zipper region at the C terminus of MSP3 may not be the only structural element responsible for oligomerization and that other peptide segments like MSP3(192–196) (YILGW) may also be required. MSP3 aggregates on incubation were transformed to long unbranched amyloid fibrils. Using immunostaining methods, we found that 5–15-μm-long fibrillar structures stained by anti-MSP3 antibodies were attached to the merozoite surface and also associated with erythrocyte membrane. We also found MSP3 to bind several molecules of heme by UV spectrophotometry, HPLC, and electrophoresis. This study suggested that its ability to bind heme is somehow related to its inherent characteristics to form oligomers. Moreover, heme interaction with a surface protein like MSP3, which does not participate in hemozoin formation, may suggest a protective role against the heme released from unprocessed hemoglobin released after schizont egress. These studies point to the other roles that MSP3 may play during the blood stages of the parasite, in addition to be an important vaccine candidate.

Plasma is the fourth state of matter and other states of matter are liquid, gas, and solid 4 . Plasma occurs as a natural phenomenon in the universe in the form of fire, in the polar aurora borealis and in the nuclear fusion reactions of the sun and also can be created artificially which has gained importance in the fields of plasma screens or light sources 1 . There are two types of plasma: thermal and non-thermal or cold atmospheric plasma. Thermal plasma has electrons and heavy particles (neutral and ions) at the same temperature. Cold Atmospheric Plasma (CAP) is said to be non-thermal because it has electron at a hotter temperature than the heavy particles that are at room temperature. . Cold Atmospheric Plasma is a specific type of plasma that is less than 104°F at the point of application 4 . So the bright future of dentistry with help of cold plasma. Key word:- cold plasma, non thermal atmospheric plasma

The mandibular first molars ( primary and permanent) usually have 2 roots located mesially and distally, but occasionally they have an additional root located distolingually requiring special attention when root canal treatment is being considered . The additional root is regarded as a normal racial and morphological variation rather than as an abnormality. It appears to be more common in races of Mongoloid origin, including Malay, Chinese, Japanese, Eskimo, and American and Canadian Indian populations. 1 To study about the rarest root and root canal anatomy using spiral ct and r.v.g. Computed tomography and radio visiography has long been used as a diagnostic aid in dentistry .With the advent of new diagnostic aids like spiral computed tomography it has become very easy for the dentist to diagnose different root and root canal anatomy and accordingly plan the treatment.

Plasmodium falciparum invades host erythrocytes by multiple invasion pathways. The invasion of erythrocytes by P. falciparum merozoites is a complex process that requires multiple interactions between host receptors and parasite ligands. A number of parasite proteins that mediate interaction with host receptors during invasion are localized to membrane-bound apical organelles referred to as micronemes and rhoptries. The timely release of these proteins to the merozoite surface is crucial for receptor engagement and invasion. It has been demonstrated previously that exposure of merozoites to a low potassium (K(+)) ionic environment as found in blood plasma leads to a rise in cytosolic calcium (Ca(2+)), which triggers microneme secretion. The signalling pathways that regulate microneme discharge in response to rise in cytosolic Ca(2+) are not completely understood. Here, we show that a P. falciparum Ca(2+)-dependent protein phosphatase, calcineurin (PfCN), is an essential regulator of Ca(2+)-dependent microneme exocytosis. An increase in PfCN activity was observed in merozoites following exposure to a low K(+) environment. Treatment of merozoites with calcineurin inhibitors such as FK506 and cyclosporin A prior to transfer to a low K(+) environment resulted in inhibition of secretion of microneme protein apical merozoite antigen-1 (PfAMA-1). Inhibition of PfCN was shown to result in reduced dephosphorylation and depolymerization of apical actin, which appears to be criticalfor microneme secretion. PfCN thus serves as an effector of Ca(2+)-dependent microneme exocytosis by regulating depolymerization of apical actin. Inhibitors that target PfCN block microneme exocytosis and limit growth of P. falciparum blood-stage parasites providing a novel approach towards development of new therapeutic strategies against malaria.

The present study attempted to find out the relationship between positive and negative clinical symptoms and various attentional task impairment in a group of schizophrenic patients. METHODS: Fifty schizophrenic patients were assessed using the Positive and Negative Syndrome Scale (PANSS) by a trained psychiatrist (TKA) who was blind to attentional test measures and two groups, each of 25 positive symptom and 25 negative symptom schizophrenic patients, were formed. On these 50 patients with schizophrenia and 15 normal control groups, various attentional test measures were applied by a clinical psychologist (SS) who remained blind to the PANSS score. RESULTS: It was found that schizophrenic patients were deficient in performing simple auditory and visual attentional tasks in comparison to normal subjects. The results of this study are inconsistent with the assumption that deficits in attention are uniquely associated with negative symptoms. The findings clearly support the hypothesis of a relationship between type of attentional processing and “dimensions” of schizophrenic symptomatology. The positive symptoms patients seem to be associated with attentional dysfunction especially selective attention and short term recall, whereas negative symptoms patients seem to be associated with different types of attentional deficits, e.g., sustained attention and visual attention. CONCLUSIONS: The findings of our study are consistent with the existing literature that schizophrenic patients in general perform poorly on various measures of attentional tasks. Positive and negative symptoms schizophrenics have some correlation with distinct attentional deficits.DOI: http://dx.doi.org/10.3126/jucms.v1i2.8402 Journal of Universal College of Medical Sciences Vol.1(2) 2013: 1-7

Summary Host cell invasion by Plasmodium falciparum requires multiple molecular interactions between host receptors and parasite ligands. A family of parasite proteins, which contain the conserved thrombospondin structural repeat motif (TSR), has been implicated in receptor binding during invasion. In this study we have characterized the functional role of a TSR containing blood stage protein referred to as P. falciparum thrombospondin related apical merozoite protein (PfTRAMP). Both native and recombinant PfTRAMP bind untreated as well as neuraminidase, trypsin or chymotrypsin-treated human erythrocytes. PfTRAMP is localized in the rhoptry bulb and is secreted during invasion. Adhesion of microneme protein EBA175 with its erythrocyte receptor glycophorin A provides the signal that triggers release of PfTRAMP from the rhoptries. Rabbit antibodies raised against PfTRAMP block erythrocyte invasion by P. falciparum suggesting that PfTRAMP plays an important functional role in invasion. Combination of antibodies against PfTRAMP with antibodies against microneme protein EBA175 provides an additive inhibitory effect against invasion. These observations suggest that targeting multiple conserved parasite ligands involved in different steps of invasion may provide an effective strategy forthe development of vaccines against blood stage malaria parasites.

The clinical symptoms of malaria are attributed to the blood stage life cycle of parasite in which merozoite invades erythrocyte, undergoes multiplication and exit to re-invade into new erythrocyte to continue its life cycle. The interaction of repertoire of parasite proteins with host cell receptors is essential for invasion process. Identification, characterization and localization of the proteins involved in invasion will enrich our understanding of this complex process. In the present study we have identified a novel Apical Rhoptry Neck Protein in Plasmodium falciparum, which harbours a predicted signal and transmembrane domain and is conserved across the species. The transcription and translation analysis confirmed its expression in schizont stage of asexual cycle of P. falciparum. Immunoflouresence microscopy in schizonts and merozoites revealed its localization in the neck of rhoptries of P. falciparum. Furthermore, PfARNP has been found at the tight junction during invasion of P. falciparum merozoite to erythrocyte.