Malik Zainul Abdin

Concentration-dependent toxicity of iron oxide nanoparticles mediated by increased oxidative stress

Iron oxide nanoparticles with unique magnetic properties have a high potential for use in several biomedical, bioengineering and in vivo applications, including tissue repair, magnetic resonance imaging, immunoassay, drug delivery, detoxification of biologic fluids, cell sorting, and hyperthermia. Although various surface modifications are being done for making these nonbiodegradable nanoparticles more biocompatible, their toxic potential is still a major concern. The current in vitro study of the interaction of superparamagnetic iron oxide nanoparticles of mean diameter 30 nm coated with Tween 80 and murine macrophage (J774) cells was undertaken to evaluate the dose- and time-dependent toxic potential, as well as investigate the role of oxidative stress in the toxicity. A 15–30 nm size range of spherical nanoparticles were characterized by transmission electron microscopy and zeta sizer. MTT assay showed >95% viability of cells in lower concentrations (25–200 μg/mL) and up to three hours of exposure, whereas at higher concentrations (300–500 μg/mL) and prolonged (six hours) exposure viability reduced to 55%–65%. Necrosis-apoptosis assay by propidium iodide and Hoechst-33342 staining revealed loss of the majority of the cells by apoptosis. H2DCFDDA assay to quantify generation of intracellular reactive oxygen species (ROS) indicated that exposure to a higher concentration of nanoparticles resulted in enhanced ROS generation, leading to cell injury and death. The cell membrane injury induced by nanoparticles studied using the lactate dehydrogenase assay, showed both concentration- and time-dependent damage. Thus, this study concluded that use of a low optimum concentration of superparamagnetic iron oxide nanoparticles is important for avoidance of oxidative stress-induced cell injury and death.

Chitinases: An update

Chitin, the second most abundant polysaccharide in nature after cellulose, is found in the exoskeleton of insects, fungi, yeast, and algae, and in the internal structures of other vertebrates. Chitinases are enzymes that degrade chitin. Chitinases contribute to the generation of carbon and nitrogen in the ecosystem. Chitin and chitinolytic enzymes are gaining importance for their biotechnological applications, especially the chitinases exploited in agriculture fields to control pathogens. Chitinases have a use in human health care, especially in human diseases like asthma. Chitinases have wide-ranging applications including the preparation of pharmaceutically important chitooligosaccharides and N-acetyl D glucosamine, preparation of single-cell protein, isolation of protoplasts from fungi and yeast, control of pathogenic fungi, treatment of chitinous waste, mosquito control and morphogenesis, etc. In this review, the various types of chitinases and the chitinases found in different organisms such as bacteria, plants, fungi, and mammals are discussed.

Lead-induced oxidative stress and metabolic alterations in Cassia angustifolia Vahl.

Forty-five-days old plants of Indian senna (Cassia angustifolia Vahl.) were subjected to 0–500 µM lead acetate (Pb-Ac) in pot culture. Changes in contents of thiobarbituric acid reactive substances (TBARS), ascorbate, glutathione, proline, sennosides (a+b), and activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), glutathione reductase (GR), and catalase (CAT) were studied at pre-flowering (60 d after sawing, DAS), flowering (90 DAS) and post-flowering (120 DAS) stages of plant development. Compared with the controls, the Pb-Ac treated plants showed an increase in contents of TBARS, dehydroascorbate, oxidized and total glutathione at all stages of growth. However, sennoside yield and contents of ascorbate and reduced form of glutathione declined. Proline content increased at 60 DAS but declined thereafter. Activities of SOD, APX, GR and CAT were markedly increased. Sennoside content was higher at 60 and 90 DAS but lower at 120 DAS, compared to the control.

Overexpression of the HMG-CoA Reductase Gene Leads to Enhanced Artemisinin Biosynthesis in Transgenic Artemisia annua Plants

An effective and affordable treatment against malaria is still a challenge for medicine. Most contemporary drugs either are too expensive to produce or are not effective against resistant strains of the malaria parasite Plasmodium falciparum. The plant Artemisia annua L. is the source of artemisinin, an effective drug against malaria for which no resistant strains of the bacterium have been reported. However, the artemisinin content of A. annua is very low, which makes its production expensive. Here we report the use of transgenic technology to increase the artemisinin content of A. annua. We report the production of transgenic plants of A. annua into which we transferred 3-hydroxy-3-methylglutaryl CoA reductase (HMGR) gene from Catharanthus roseus (L.) G. Don using Agrobacterium-mediated gene transfer technology. Transgene integration and copy number were assessed by PCR and Southern hybridization, which confirmed the stable integration of multiple copies of the transgene in 7 different transgenic lines of A. annua. The leaf tissue of three of the A. annua transgenic lines possessed significantly higher HMGR activity compared with wild-type controls, and this activity was associated exclusively with microsomal membranes. The artemisinin content of the shoots of one of the transgenic lines depicted an increase of 22.5 % artemisinin content compared with wild-type control A. annua plants.

Identification and bioactive potential of endophytic fungi isolated from selected plants of the Western Himalayas

This study was conducted to characterize and explore the endophytic fungi of selected plants from the Western Himalayas for their bioactive potential. A total of 72 strains of endophytic fungi were isolated and characterized morphologically as well as on the basis of ITS1-5.8S-ITS2 ribosomal gene sequence acquisition and analyses. The fungi represented 27 genera of which two belonged to Basidiomycota, each representing a single isolate, while the rest of the isolates comprised of Ascomycetous fungi. Among the isolated strains, ten isolates could not be assigned to a genus as they displayed a maximum sequence similarity of 95% or less with taxonomically characterized organisms. Among the host plants, the conifers, Cedrus deodara, Pinus roxburgii and Abies pindrow harbored the most diverse fungi, belonging to 13 different genera, which represented almost half of the total genera isolated. Several extracts prepared from the fermented broth of these fungi demonstrated strong bioactivity against E. coli and S. aureus with the lowest IC50 of 18 μg/ml obtained with the extract of Trichophaea abundans inhabiting Pinus sp. In comparison, extracts from only three endophytes were significantly inhibitory to Candida albicans, an important fungal pathogen. Further, 24 endophytes inhibited three or more phytopathogens by at least 50% in co-culture, among a panel of seven test organisms. Extracts from 17 fungi possessed immuno-modulatory activities with five of them showing significant immune suppression as demonstrated by the in vitro lymphocyte proliferation assay. This study is an important step towards tapping the endophytic fungal diversity from the Western Himalayas and assessing their bioactive potential. Further studies on the selected endophytes may lead to the isolation of novel natural products for use in medicine, industry and agriculture.

SCAR markers: a potential tool for authentication of herbal drugs.

Randomly Amplified Polymorphic DNA (RAPD) is easy to develop and simple molecular marker, but lack of reproducibility makes it less reliable for authentication of herbal drugs. Besides RAPD, other popular PCR and non-PCR based markers like AFLP, ISSR, SSR and RFLP are also used for authentication. However, these also have disadvantages like use of radioactive isotopes, high cost and absolute requirement of sequence information. Therefore, it is a better option to improve the reproducibility of RAPD by converting RAPD amplicons into Sequence Characterized Amplified Region (SCAR) markers. SCAR markers are easy, reliable and reproducible thus, have an advantage over RAPD markers for authentication of medicinal herbs used in the preparation of traditional medicines. These markers however, have been developed for only a few medicinal herbs. This review is an attempt to evaluate critically the role of SCAR markers in authentication of medicinal herbs used in traditional formulations.

Ameliorative Effects of CaCl2 on Growth, Ionic Relations, and Proline Content of Senna Under Salinity Stress

ABSTRACT The effect of NaCl and CaCl2 on growth parameters, ionic relations, and proline level in senna (Cassia angustifolia) plant was investigated in a pot culture experiment using NaCl (80 mM and 160 mM), CaCl2 (5 mM and 10 mM), and the combined salt of NaCl + CaCl2 (80 mM + 10 mM and 160 mM + 10 mM). These treatments were given to the growing plant separately at pre-flowering (A1), flowering (A2) and post-flowering (A3) stages. Observations were recorded at 15-day intervals from the time of treatments till harvest. Compared to the controls, biomass of the root, shoot, and leaf decreased significantly with each NaCl treatment, whereas the reverse was observed with CaCl2 treatments. Combined treatments of NaCl + CaCl2 applied at different stages reduced the biomass, but this reduction was less than the one observed with NaCl treatments alone. However, proline accumulation in the leaves was 8 times higher than in the controls with treatment of 160 mM NaCl + 10 mM CaCl2, whereas it was 5 times higher with NaCl (160 mM) alone. The concentration of K+ and Ca2+ was inhibited with NaCl treatments, while Na+ and Cl− levels increased in the different plant parts. CaCl2 treatment enhanced the K+ and Ca2+ concentration, while the combined treatments mitigated the adverse effect caused by NaCl. Thus calcium could alleviate the NaCl-induced inhibition of plant growth via the maintenance of net K+ to Na+ selectivity and the enhancement of proline accumulation in the leaves.

Growth and Metabolism of Senna as Affected by Salt Stress

Pot culture experiments were conducted using different NaCl concentrations to assess their impact on the growth and metabolic changes in senna (Cassia angustifolia Vahl.). Five treatments (0, 40, 80, 120, and 160 mM NaCl) were given to the plants at three phenological stages, i.e. at pre-flowering, (45 days after sowing, DAS); flowering (75 DAS) and post-flowering (90 DAS) stages. A significant reduction in the biomass and length of the roots and shoots, photosynthetic rate, stomatal conductance, the total chlorophyll content, protein content, nitrate reductase activity, and reduced nitrogen content of the leaves was observed at each phenological stage with each salt concentration applied. Contrary to this, proline and nitrate contents of the leaves increased markedly. The post-flowering stage was most sensitive to NaCl treatment.

Enhancement of artemisinin content by constitutive expression of the HMG-CoA reductase gene in high-yielding strain of Artemisia annua L.

Artemisinin is effective against both chloroquine-resistant and -sensitive strains of Plasmodium species. However, the low yield of artemisinin from cultivated and wild plants is a serious limitation to the commercialization of this drug. Optimization of artemisinin yield either in vivo or in vitro is therefore highly desirable. To this end, we have overexpressed the 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase (HMGR) gene (hmgr) from Catharanthusroseus L. in Artemisia annua L. and analyzed its influence on artemisinin content. PCR and Southern blot analyses revealed that the transgenic plants showed stable integration of the foreign hmgr gene. The reverse transcriptase-PCR results suggested that the hmgr was expressed at the transcriptional level in transgenic lines of Artemisia annua L., while the high-performance liquid chromatography analysis showed that artemisinin content was significantly increased in a number of the transgenic lines. Artemisinin content in one of the A. annua transgenic lines was 38.9% higher than that in non-transgenic plants, and HMGR enzyme activity in transgenic A. annua L. was also higher than that in the non-transgenic lines.

Effect of split application of sulphur and nitrogen on growth and yield attributes of Brassica genotypes differing in time of flowering

According to prevalent agronomic practices for cultivation of Brassica genotypes, N is applied in split doses, while S is applied as a basal dose. This may create imbalance in the supply of these nutrients during the growth and development of the crop because metabolism of N and that of S are closely linked and play a central role in protein synthesis. The requirement of one depends on the supply of the other, and the imbalance in their supply causes a reduction in the yield because of reduced uptake and assimilation of the two nutrients. In the present investigation, therefore, S was applied in split doses, along with N, to study its effect on growth and yield attributes of Brassica juncea (L.) Czern. and Coss. (V1) and Brassica campestris L. (V2). In the experiment, conducted in the field, 40 kg S ha−1 as CaSO4 (gypsum) was applied either in a single basal application (S1) or in two (S2) or three (S3) split applications; and 100 kg N ha−1 as urea was applied either in two (N2) or three (N3) splits. Biom...