Exploring novel potential of mycosynthesized magnetic nanoparticles for phosphatase immobilization and biological activity.

Among different microbes, fungi are proficient candidates for the extracellular synthesis of iron nanoparticles. For biogenic synthesis of iron nanoparticles, a thermophilic mould Myceliophthora thermophila BJTLRMDU7 was used in this study. Mycogenic magnetic nanoparticles were used for phosphatase immobilization and therapeutic applications such as antimicrobial and antimalarial activity. Firstly, the phosphatase was immobilized on biogenic iron nanoparticles with an efficiency of >56 %. Immobilized enzyme was optimally active at 60 °C and pH 5. Immobilized phosphatase was recycled using external magnetic field up to 4th cycle retaining >50 % activity. The immobilized phosphatase efficiently released inorganic phosphate from different flours such as wheat, maize and gram at 37 °C and 60 °C. There was continuous increase in the release of inorganic phosphorus from all samples with incubation time at 37 °C and slight reduction at 60 °C. These nanoparticles showed the effective antimicrobial activity against Bacillus subtilis, Escherichia coli and Myceliophthora thermophila. Further, the synthesized iron nanoparticles showed antimalarial potential against Plasmodium falciparum. Biogenic nanoparticles did not exhibit hemolytic activity and cytotoxicity. Therefore, biogenic iron nanoparticles could be used as a suitable matrix for immobilization of enzymes and safe therapeutics.

Antimalarial potential of Kerala Ayurvedic Water.

BACKGROUND OBJECTIVES: The persistent threat of drug resistant malaria demands new cures. Low prevalence of malaria in the Indian state of Kerala compared with other proximal states made us explore if there is any traditional practice in Kerala which may confer protection against malaria. In this context, our attention was drawn to 'Pathimugam' i.e., Ceasalpinia sappan whose heartwood is used to prepare a red aqueous extract which is a uniquely popular drink in Kerala. METHODS: Aqueous and methanolic extracts of various organs of C. sappan were prepared and tested against Plasmodium falciparum grown in vitro culture using SYBR Green-I assay. The cytotoxicity of active extracts/fractions was studied using mammalian HeLa cell line. in vivo efficacy was determined using P. berghei ANKA infected mice. RESULTS: The highest antiplasmodial activities in the alcoholic and aqueous extracts were observed in leaf methanolic extract (IC50 2 µg/ml) and heartwood aqueous extract (IC50 12.5 µg/ml). Ceasalpinia sappan extracts were equipotent against both chloroquine-sensitive Pf3D7 and resistant PfINDO strains and showed suppression of percentage parasitemia in P. berghei infected mice. Activity- guided chromatographic fractionation of aqueous wood extract led to the fortification of antiplasmodial activity (IC50 5 µg/ml). INTERPRETATION CONCLUSION: Our results establish the antiplasmodial potential of C. sappan and suggest that its regular use might have prophylactic or curative actions that may assist in keeping check on malaria in the Indian state of Kerala.

Vaccine development: Current trends and technologies.

Despite the effectiveness of vaccination in reducing or eradicating diseases caused by pathogens, there remain certain diseases and emerging infections for which developing effective vaccines is inherently challenging. Additionally, developing vaccines for individuals with compromised immune systems or underlying medical conditions presents significant difficulties. As well as traditional vaccine different methods such as inactivated or live attenuated vaccines, viral vector vaccines, and subunit vaccines, emerging non-viral vaccine technologies, including viral-like particle and nanoparticle vaccines, DNA/RNA vaccines, and rational vaccine design, offer new strategies to address the existing challenges in vaccine development. These advancements have also greatly enhanced our understanding of vaccine immunology, which will guide future vaccine development for a broad range of diseases, including rapidly emerging infectious diseases like COVID-19 and diseases that have historically proven resistant to vaccination. This review provides a comprehensive assessment of emerging non-viral vaccine production methods and their application in addressing the fundamental and current challenges in vaccine development.

Green synthesis of iron nanoparticles: Sources and multifarious biotechnological applications.

Green synthesis of iron nanoparticles is a highly fascinating research area and has gained importance due to reliable, sustainable and ecofriendly protocol for synthesizing nanoparticles, along with the easy availability of plant materials and their pharmacological significance. As an alternate to physical and chemical synthesis, the biological materials, like microorganisms and plants are considered to be less costly and environment-friendly. Iron nanoparticles with diverse morphology and size have been synthesized using biological extracts. Microbial (bacteria, fungi, algae etc.) and plant extracts have been employed in green synthesis of iron nanoparticles due to the presence of various metabolites and biomolecules. Physical and biochemical properties of biologically synthesized iron nanoparticles are superior to that are synthesized using physical and chemical agents. Iron nanoparticles have magnetic property with thermal and electrical conductivity. Iron nanoparticles below a certain size (generally 10-20 nm), can exhibit a unique form of magnetism called superparamagnetism. They are non-toxic and highly dispersible with targeted delivery, which are suitable for efficient drug delivery to the target. Green synthesized iron nanoparticles have been explored for multifarious biotechnological applications. These iron nanoparticles exhibited antimicrobial and anticancerous properties. Iron nanoparticles adversely affect the cell viability, division and metabolic activity. Iron nanoparticles have been used in the purification and immobilization of various enzymes/proteins. Iron nanoparticles have shown potential in bioremediation of various organic and inorganic pollutants. This review describes various biological sources used in the green synthesis of iron nanoparticles and their potential applications in biotechnology, diagnostics and mitigation of environmental pollutants.

Linking population-based cohorts with cancer registries in LMIC: a case study and lessons learnt in India.

OBJECTIVES: In resource-constrained settings, cancer epidemiology research typically relies on self-reported diagnoses. To test a more systematic alternative approach, we assessed the feasibility of linking a cohort with a cancer registry. SETTING: Data linkage was performed between a population-based cohort in Chennai, India, with a local population-based cancer registry. PARTICIPANTS: Data set of Centre for Cardiometabolic Risk Reduction in South-Asia (CARRS) cohort participants (N=11 772) from Chennai was linked with the cancer registry data set for the period 1982-2015 (N=140 986). METHODS AND OUTCOME MEASURES: Match*Pro, a probabilistic record linkage software, was used for computerised linkages followed by manual review of high scoring records. The variables used for linkage included participant name, gender, age, address, Postal Index Number and father's and spouse's name. Registry records between 2010 and 2015 and between 1982 and 2015, respectively, represented incident and all (both incident and prevalent) cases. The extent of agreement between self-reports and registry-based ascertainment was expressed as the proportion of cases found in both data sets among cases identified independently in each source. RESULTS: There were 52 self-reported cancer cases among 11 772 cohort participants, but 5 cases were misreported. Of the remaining 47 eligible self-reported cases (incident and prevalent), 37 (79%) were confirmed by registry linkage. Among 29 self-reported incident cancers, 25 (86%) were found in the registry. Registry linkage also identified 24 previously not reported cancers; 12 of those were incident cases. The likelihood of linkage was higher in more recent years (2014-2015). CONCLUSIONS: Although linkage variables in this study had limited discriminatory power in the absence of a unique identifier, an appreciable proportion of self-reported cases were confirmed in the registry via linkages. More importantly, the linkages also identified many previously unreported cases. These findings offer new insights that can inform future cancer surveillance and research in low-income and middle-income countries.

Antiplasmodial activity of the bulbs of Fritillaria cirrhosa D.Don (Syn: Fritillaria roylei Hook.): UPLC-IM-Q-TOF-MS/MS-based biochemometric approach for the identification of marker compounds.

ETHNOPHARMACOLOGICAL RELEVANCE: Fritillaria cirrhosa D.Don (Syn: Fritillaria roylei Hook.) (Hindi name: Kshirakakoli) is a critically endangered Himalayan medicinal plant, well documented in Ayurveda for its therapeutic uses against various disorders such as jvara (fever), kasa (respiratory tract disease) etc. Its bulbs are also used as Szechuan-Pei-Mu for their antipyretic properties in the traditional Chinese medicine. However, despite its ethnomedicinal usage, the therapeutic use of F. cirrhosa bulbs for jvara (fever) related conditions such as malaria has remained unexplored. Hence in the context of increasing global concerns about drug-resistant malaria, it is important to investigate the antiplasmodial activity of F. cirrhosa bulbs for novel antimalarial agents. AIM OF THE STUDY: To investigate the antiplasmodial effects of the extracts/fractions of F. cirrhosa bulbs by the biochemometric approach and to rationalize its ethnopharmacological usage for jvara (fever) related conditions such as malaria. MATERIAL AND METHODS: This study involves the UHPLC-MS-based plant material selection, preparation, quantification, and assessment of F. cirrhosa bulb extracts against CQ-sensitive Pf 3D7 & CQ-resistant Pf INDO strains. Further, UPLC-IM-Q-TOF-MS-based biochemometric approach has been applied for the identification of marker compounds responsible for the observed antiplasmodial effects. The identified marker compounds were also assessed for their in silico ADMET properties and binding efficacy with the drug transporter Pf CRT. RESULTS: Different F. cirrhosa bulb extracts/fractions showed promising antiplasmodial activity with IC50 values 2.71-19.77 µg/mL for CQ-resistant Pf INDO strain and 1.76-21.52 µg/mL for CQ-sensitive Pf 3D7 strain. UPLC-IM-Q-TOF-MS/MS-based biochemometric analysis revealed four marker compounds i.e., peimine (m/z 432.3448), peimisine (m/z 428.3504), puqiedinone (m/z 414.3379), and puqiedine (m/z 416.3509) responsible for the observed antiplasmodial activity. The identified marker compounds showed excellent binding efficacy with Pf CRT and suitable drug-like properties in silico. CONCLUSIONS: The study demonstrated promising antiplasmodial activity of the chloroform and alkaloid enriched fractions of F. cirrhosa bulbs and further identified the four marker compounds responsible for the promising antiplasmodial activity. These marker compounds i.e., peimine, peimisine, puqiedinone and puqiedine were identified by the biochemometric analysis as the putative antiplasmodial constituents of the F. cirrhosa bulbs. Further, in silico studies indicated the good binding affinity of the marker compounds with Pf CRT along with suitable ADMET properties. Overall, the study elucidates the antiplasmodial activity of F. cirrhosa bulbs from the western Himalayan region and provides nascent scientific evidence for their ethnopharmacological usage in jvara (fever) related conditions such as malaria.

Exploration of Anti-plasmodial Activity of Prunus cerasoides Buch.-Ham. ex D. Don (family: Rosaceae) and Its Wood Chromatographic Fractions.

PURPOSE: Increasing resistance to the currently available antimalarial drugs is a leading cause of failure to control malaria. Plant-based medicines are commonly used to manage numerous infections, making medicinal plants the best possible source of alternative antimalarial drugs. The objective of this study is therefore to identify antimalarial potential of Prunus cerasoides. METHODS: Here, anti-plasmodial activity of crude methanolic and aqueous extracts of Prunus cerasoides and fractions obtained by reverse-phase high performance liquid chromatography (RPHPLC) were tested for in vitro activity against chloroquine sensitive Plasmodium falciparum 3D7 and chloroquine resistant INDO & Dd2 strains using SYBR Green I assay. The cytotoxic activity of active extracts/fractions was evaluated against mammalian cell lines-HeLa using MTT assay. RESULTS: Aqueous extracts of leaves, wood, bark and fruit of P. cerasoides showed poor to no activity up to 100 µg/ml, however methanolic extract showed moderate (IC50: 21-60 µg/ml) to poor (IC50: 61-100 µg/ml) anti-plasmodial activity. Fractionation of wood methanolic extract led to enrichment in antimalarial activity in some of its fractions as out of 17 fractions collected, good anti-plasmodial activity (IC50: 1-20 µg/ml) was shown by three fractions and nine fractions showed moderate anti-plasmodial activity. However, five fractions showed poor to no activity against Plasmodium falciparum (IC50:61-100 and >100 µg/ml respectively). Furthermore, these active fractions showed no cytotoxic effects on mammalian cell lines. CONCLUSION: Findings of this study elucidate the anti-plasmodial potential of P. cerasoides and validate its traditional usage suggesting that it could be a possible source of a drug candidate in combating this disease.

Assessment of in vitro and in vivo antimalarial efficacy and GC-fingerprints of selected medicinal plant extracts.

A hallmark of mortality and morbidity, malaria is affecting nearly half of the world's population. Emergence of drug-resistant strains of malarial parasite prompts identification and evaluation of medicinal plants and their constituents that may hold the key to a new and effective anti-malarial drug. In this context, nineteen methanolic extracts from seventeen medicinal plants were evaluated for anti-plasmodial potential against Plasmodium falciparum strain 3D7 (Chloroquine (CQ) sensitive) and INDO (CQ resistant) using fluorescence based SYBR-Green assay and for cytotoxic effects against mammalian cell lines. Leaf extract of two plants showed promising in vitro anti-malarial activity (Pf3D7 IC50 ≤ 10 µg/ml); one plant extract showed good activity (Pf3D7 IC50 = 10.1-20 µg/ml); seven were moderately active (IC50 = 20.1-50 µg/ml), four plant extracts showed poor activity (PfD7 IC50 = 50.1-100 µg/ml) and five extracts showed no activity up to IC50 = 100 µg/ml. Further, six extracts were found equipotent to PfINDO (resistance index ranging 0.4-2) and relatively nontoxic to mammalian cell lines HEK293 (cytotoxicity index ranging 1.4-12.5). Based on good resistance and selectivity indices, three extracts were evaluated for in vivo activity in Plasmodium berghei ANKA infected mice at a dose of 500 mg/kg and they showed significant suppression of P. berghei parasitemia. Further, these active plant extracts were fractionated using silica-gel chromatography and their fractions were evaluated for anti-plasmodial action. Obtained fractions showed enrichment in antimalarial activity. Active fractions were analyzed by gas chromatography and mass-spectrometery. Results suggests that the three active plant extracts could serve as potent source of anti-malarial agent and therefore require further analysis.

In silico Potential of Approved Antimalarial Drugs for Repurposing Against COVID-19.

Although the coronavirus disease 2019 (COVID-19) pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is wreaking havoc and resulting in mortality and morbidity across the planet, novel treatments are urgently needed. Drug repurposing offers an innovative approach in this context. We report here new findings on the in silico potential of several antimalarial drugs for repurposing against COVID-19. We conducted analyses by docking the compounds against two SARS-CoV-2-specific targets: (1) the receptor binding domain spike protein and (2) the main protease of the virus (MPro) using the Schrödinger software. Importantly, the docking analysis revealed that doxycycline (DOX) showed the most effective binding to the spike protein of SARS-CoV-2, whereas halofantrine and mefloquine bound effectively with the main protease among the antimalarial drugs evaluated in the present study. The in silico approach reported here suggested that DOX could potentially be a good candidate for repurposing for COVID-19. In contrast, to decipher the actual potential of DOX and halofantrine against COVID-19, further in vitro and in vivo studies are called for. Drug repurposing warrants consideration as a viable research and innovation avenue as planetary health efforts to fight the COVID-19 continue.

Dysbiosis Disrupts Gut Immune Homeostasis and Promotes Gastric Diseases.

Perturbation in the microbial population/colony index has harmful consequences on human health. Both biological and social factors influence the composition of the gut microbiota and also promote gastric diseases. Changes in the gut microbiota manifest in disease progression owing to epigenetic modification in the host, which in turn influences differentiation and function of immune cells adversely. Uncontrolled use of antibiotics, chemotherapeutic drugs, and any change in the diet pattern usually contribute to the changes in the colony index of sensitive strains known to release microbial content in the tissue micromilieu. Ligands released from dying microbes induce Toll-like receptor (TLR) mimicry, skew hypoxia, and cause sterile inflammation, which further contributes to the severity of inflammatory, autoimmune, and tumorous diseases. The major aim and scope of this review is both to discuss various modalities/interventions across the globe and to utilize microbiota-based therapeutic approaches for mitigating the disease burden.

Multilayer ReS2 Photodetectors with Gate Tunability for High Responsivity and High-Speed Applications.

Rhenium disulfide (ReS2) is an attractive candidate for photodetection applications owing to its thickness-independent direct band gap. Despite various photodetection studies using two-dimensional semiconductors, the trade-off between responsivity and response time under varying measurement conditions has not been studied in detail. This report presents a comprehensive study of the architectural, laser power and gate bias dependence of responsivity and speed in supported and suspended ReS2 phototransistors. Photocurrent scans show uniform photogeneration across the entire channel because of enhanced optical absorption and a direct band gap in multilayer ReS2. A high responsivity of 4 A W-1 (at 50 ms response time) and a low response time of 20 µs (at 4 mA W-1 responsivity) make this one of the fastest reported transition-metal dichalcogenide photodetectors. Occupancy of intrinsic (bulk ReS2) and extrinsic (ReS2/SiO2 interface) traps is modulated using gate bias to demonstrate tunability of the response time (responsivity) over 4 orders (15×) of magnitude, highlighting the versatility of these photodetectors. Differences in the trap distributions of suspended and supported channel architectures, and their occupancy under different gate biases enable switching the dominant operating mechanism between either photogating or photoconduction. Further, a new metric that captures intrinsic photodetector performance by including the trade-off between its responsivity and speed, besides normalizing for the applied bias and geometry, is proposed and benchmarked for this work.

Enhanced stability and performance of few-layer black phosphorus transistors by electron beam irradiation.

Few layer black phosphorus (BP) has recently emerged as a potential graphene analogue due to its high mobility and direct, appreciable, band gap. The fabrication and characterization of field effect transistors (FETs) involves exposure of the channel material to an electron beam (e-beam) in imaging techniques such as transmission electron microscopy (TEM) and scanning electron microscopy (SEM), and fabrication techniques like electron beam lithography (EBL). Despite this, the effect of e-beam irradiation on BP-FET performance has not been studied experimentally. In this work, we report the first experimental study on the impact of e-beam irradiation on BP-FETs. An electron beam is known to induce defects and structural changes in 2D materials like graphene, MoS2etc. resulting in the deterioration of the device quality. However, for BP-FETs, we observe an improvement in the on-current and carrier mobility (µ) along with a decrease in threshold voltage (Vth) on exposure to an e-beam with 15 keV energy for 80 seconds. These changes can be attributed to the capture of electrons by traps near the SiO2-BP interface and reduced BP surface roughness due to e-beam exposure. Hysteresis measurements and physical characterization (i.e. atomic force microscopy (AFM), X-ray photoelectron (XPS) and Raman spectroscopies) validate these mechanisms. Reduced hysteresis indicates occupation of the traps, AFM surface scans indicate reduced surface roughness and XPS data show a reduced phosphorus oxide (POx) peak immediately after exposure. Raman measurements indicate a probable structural change due to the interaction between e-beam and BP which could result in better stability.

Exploration of antiplasmodial activity in Acalypha wilkesiana Müller Argoviensis, 1866 (family: Euphorbiaceae) and its GC-MS fingerprint.

The plant kingdom continues to hold great promise for the eradication of Malaria infection following the challenges of insecticide resistance by the vector mosquito, drug resistance by the parasite, and the development of a vaccine still being a mirage. Acalypha wilkesiana Muller Argoviensis, 1866 (family: Euphorbiaceae) leaves have the ethnopharmacological reputation for use as a remedy against dermal microbial infections in Nigeria. Here, we have studied the antiplasmodial potential of the extract of the leaves of this ornamental plant. Aqueous methanol crude extract (70%) and Prep reversed-phase high-performance liquid chromatography (RPHPLC) fractions were tested in vitro against blood stage Plasmodium falciparum 3D7 strain parasites for antiplasmodial activity using the SYBR Green assay. Results obtained were validated through Giemsa stained microscopic blood smeared slides. An IC50 of < 0.39 µg/ml for fractions of the RPHPLC together with TC50 of > 100 µg/ml against mammalian HUH-7 cell lines and a HC50 of > 100 µg/ml against red blood cells indicate a high selectivity of this plant against Plasmodium. This is the first report of the antiplasmodial activity of this plant and a GC-MS fingerprinting of the same, opening the possibilities of identifying novel pharmacophores against the malaria parasite.

Few-Layer MoS2 p-Type Devices Enabled by Selective Doping Using Low Energy Phosphorus Implantation.

P-type doping of MoS2 has proved to be a significant bottleneck in the realization of fundamental devices such as p-n junction diodes and p-type transistors due to its intrinsic n-type behavior. We report a CMOS compatible, controllable and area selective phosphorus plasma immersion ion implantation (PIII) process for p-type doping of MoS2. Physical characterization using SIMS, AFM, XRD and Raman techniques was used to identify process conditions with reduced lattice defects as well as low surface damage and etching, 4X lower than previous plasma based doping reports for MoS2. A wide range of nondegenerate to degenerate p-type doping is demonstrated in MoS2 field effect transistors exhibiting dominant hole transport. Nearly ideal and air stable, lateral homogeneous p-n junction diodes with a gate-tunable rectification ratio as high as 2 × 10(4) are demonstrated using area selective doping. Comparison of XPS data from unimplanted and implanted MoS2 layers shows a shift of 0.67 eV toward lower binding energies for Mo and S peaks indicating p-type doping. First-principles calculations using density functional theory techniques confirm p-type doping due to charge transfer originating from substitutional as well as physisorbed phosphorus in top few layers of MoS2. Pre-existing sulfur vacancies are shown to enhance the doping level significantly.

Interfacial n-Doping Using an Ultrathin TiO2 Layer for Contact Resistance Reduction in MoS2.

We demonstrate a low and constant effective Schottky barrier height (ΦB ∼ 40 meV) irrespective of the metal work function by introducing an ultrathin TiO2 ALD interfacial layer between various metals (Ti, Ni, Au, and Pd) and MoS2. Transmission line method devices with and without the contact TiO2 interfacial layer on the same MoS2 flake demonstrate reduced (24×) contact resistance (RC) in the presence of TiO2. The insertion of TiO2 at the source-drain contact interface results in significant improvement in the on-current and field effect mobility (up to 10×). The reduction in RC and ΦB has been explained through interfacial doping of MoS2 and validated by first-principles calculations, which indicate metallic behavior of the TiO2-MoS2 interface. Consistent with DFT results of interfacial doping, X-ray photoelectron spectroscopy (XPS) data also exhibit a 0.5 eV shift toward higher binding energies for Mo 3d and S 2p peaks in the presence of TiO2, indicating Fermi level movement toward the conduction band (n-type doping). Ultraviolet photoelectron spectroscopy (UPS) further corroborates the interfacial doping model, as MoS2 flakes capped with ultrathin TiO2 exhibit a reduction of 0.3 eV in the effective work function. Finally, a systematic comparison of the impact of selective doping with the TiO2 layer under the source-drain metal relative to that on top of the MoS2 channel shows a larger benefit for transistor performance from the reduction in source-drain contact resistance.

Evaluation of antiplasmodial activity of medicinal plants from North Indian Buchpora and South Indian Eastern Ghats.

BACKGROUND: Development of resistance against the frontline anti-malarial drugs has created an alarming situation, which requires intensive drug discovery to develop new, more effective, affordable and accessible anti-malarial agents. METHODS: Inspired by their ethnobotanical reputation for being effective against febrile diseases, antiplasmodial potential of ethyl acetate extracts (EAE) and methanol extracts (ME) of 17 medicinal plants collected from the Eastern Ghats of South India and Buchpora, North India were explored against Plasmodium falciparum in vitro using the SYBR Green assay. The results were validated both by confirmation that the fall in fluorescence signal was not due to quenching effects mediated by phytochemical extracts and by Giemsa-stained microscopy. RESULTS: Using EAE or ME, promising antiplasmodial activity (IC50 Pf3D7 ≤ 20 µg/ml), was seen in Aerva lanata (Whole aerial parts-EAE), Anisomeles malabarica (Leaf-EAE), Anogeissus latifolia (bark-EAE), Cassia alata (leaves-EAE), Glycyrrhiza glabra (root-EAE), Juglans regia (seed-ME), Psidium guajava (leaf-ME and EAE) and Solanum xanthocarpum (Whole aerial parts-EAE). EAEs from leaves of Couroupita guianensis, Euphorbia hirta, Pergularia daemia, Tinospora cordifolia and Tridax procumbens as also ME from Ricinus communis (leaf and seed) showed good antiplasmodial activity (Pf 3D7 IC50 21 - 40 µg/ml). Moderate activity (Pf 3D7 IC50: 40-60 µg/mL) was shown by the leaf EAEs of Cardiospermum halicacabum, Indigofera tinctoria and Ricinus communis while the remaining extracts showed marginal (Pf 3D7 IC50 60 to >100 µg/ml) activities. The promising extracts showed good resistance indices (0.41 - 1.4) against the chloroquine resistant INDO strain of P. falciparum and good selectivity indices (3 to > 22.2) when tested against the HeLa cell line. CONCLUSION: These results provide validity to the traditional medicinal usage of some of these plants and further make a case for activity-guided purification of new pharmacophores against malaria.

Antiplasmodial activity of medicinal plants from Chhotanagpur plateau, Jharkhand, India.

BACKGROUND: The alarmingly increasing problem of drug resistance in treatment of malaria has led to an urgent need for identifying new anti-malarial drugs for both prophylaxis and chemotherapy. AIM OF THE STUDY: The present study presents a systematic exploration of the ex vivo blood stage antiplasmodial potential of medicinal plants to corroborate their traditional usage against malaria in Jharkhand, India. METHODS: An ethnobotanical survey in and around Ranchi was done to grasp the traditional knowledge of medicinal plants used by local healers for malaria, other fevers and for other medicinal purposes like, antiamoebic, antihelmenthic, antidote to poisons, etc. Following the survey, the selected 22 plant samples were extracted in ethanol for studying ex vivo SYBR Green I fluorescence assay based anti-plasmodial activity against both chloroquine-sensitive Pf3D7 and chloroquine resistant PfINDO strains of Plasmodium falciparum grown in human red blood cell cultures. Cytotoxicity was determined against HeLa and L929 cells using MTT assay. Further the most potent extract was chromatographed on reverse phase HPLC towards antiplasmodial activity guided purification of metabolites. RESULTS: Of the 22 plant species assayed, the highest antiplasmodial activity (Pf3D7IC50 ≤ 5 µg/ml) was seen in leaf ethanol extracts of Corymbia citriodora (Hook.) K.D.Hill & L.A.S.Johnson, Calotropis procera (Aiton) Dryand. and Annona squamosa L. and bark ethanol extract of Holarrhena pubescens Wall. ex G.Don. Leaf ethanol extract of H. pubescens, bark ethanol extract of Pongamia pinnata (L.) Pierre and whole plant ethanol extract of Partheniumhysterophorus L. showed promising activity (IC50 6-10 µg/ml). Good antiplasmodial activity (IC50: 11-20 µg/ml) was observed in leaf ethanol extract of Bryophyllum pinnatum (Lam.) Oken and whole plant ethanol extract of Catharanthus roseus (L.) G.Don. The extracts of plants showing highest to good antiplasmodial activity exhibited HeLa/Pf3D7 selectivity indices of the order of 20-45. Bioassay guided fractionation of P. hysterophorus led to fivefold enrichment of antiplasmodial activities (IC50 ~450 ng/ml) in some fractions. CONCLUSION: These results provide confirmation to the traditional usage of some medicinal plants against malaria in areas around Ranchi, Jharkhand.

A search for antiplasmodial metabolites among fungal endophytes of terrestrial and marine plants of southern India.

Eighty four different fungal endophytes isolated from sea grasses (5), marine algae (36) and leaves or barks of forest trees (43) were grown in vitro and the secondary metabolites secreted by them were harvested by immobilizing them on XAD beads. These metabolites were eluted with methanol and screened using SYBR Green I assay for their antiplasmodial activity against blood stage Plasmodium falciparum in human red blood cell culture. Our results revealed that fungal endophytes belonging to diverse genera elaborate antiplasmodial metabolites. A Fusarium sp. (580, IC50: 1.94 µg ml(-1)) endophytic in a marine alga and a Nigrospora sp. (151, IC50: 2.88 µg ml(-1)) endophytic in a tree species were subjected to antiplasmodial activity-guided reversed phase high performance liquid chromatography separation. Purification led to potentiation as reflected in IC50 values of 0.12 µg ml(-1) and 0.15 µg ml(-1) for two of the fractions obtained from 580. Our study adds further credence to the notion that fungal endophytes are a potential storehouse for a variety of novel secondary metabolites vested with different bioactivities including some that can stall the growth of the malaria parasite.

Plasmodium falciparum merozoite surface protein 3: oligomerization, self-assembly, and heme complex formation.

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.

Evaluation of antiplasmodial activity of green synthesized silver nanoparticles.

In the present study silver nanoparticles (silver(np)) were synthesized from AgNO3 through simple green routes using either purified Alpha Amylase or aqueous leaf extracts of Ashoka and Neem respectively. The use of plant extract/enzyme for synthesis of nanoparticles is a single-step, cost effective and eco-friendly process. The silver(np) obtained by these three different ways were characterized using UV-visible spectroscopy, DLS, TEM, XRD and FTIR. These nanoparticles were found to be antiplasmodial with IC50 (µg/ml) 3.75 (Amylase(np)), 8 (Ashoka(np)) and 30 (Neem(np)) whereas plant extracts or amylase alone did not show any activity up to 40 µg/ml. Although AgNO3 was also found to have intrinsic antiplasmodial activity (IC50 0.5 µg/ml), the hemolytic tendencies appeared to be higher for AgNO3 (MHC10: 10 µg/ml) against the nanoparticulate preparations (MHC10: >40 µg/ml).