Resveratrol-loaded chitosan-pectin core-shell nanoparticles as novel drug delivery vehicle for sustained release and improved antioxidant activities.

Resveratrol, chemically known as 3,5,4'-trihydroxy-trans-stilbene, is a natural polyphenol with promising multi-targeted health benefits. The optimal therapeutic uses of resveratrol are limited due to its poor solubility, rapid metabolism and low bioavailability. To address the issues, we have encapsulated resveratrol inside the nanosized core made of chitosan and coated this core with pectin-shell in order to fabricate a drug delivery vehicle which can entrap resveratrol for a longer period of time. The core-shell nanoparticles fabricated in this way were characterized with the help of Fourier transform infrared spectrometer, field-emission scanning electron microscope, field-emission transmission electron microscopy/selected area electron diffraction, high-resolution transmission electron microscope, dynamic light scattering and zeta potential measurements. In vitro drug release study showed the ability of the core-shell nanoparticles to provide sustained release of resveratrol for almost 30 h. The release efficiency of the drug was found to be pH dependent, and a sequential control over drug release can be obtained by varying the shell thickness. The resveratrol encapsulated in a nanocarrier was found to have a better in vitro antioxidant activity than free resveratrol as determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging method. This work finally offers a novel nano-based drug delivery system.

Green synthesis of silver nanoplates using the special category of plant leaves showing the lotus effect.

This paper reports the first ever green synthesis of silver nanoplates using plant leaves having the special feature of showing the lotus effect. Eichhornia crassipes and Colocasia esculenta plant leaves were chosen for the purpose. The aqueous leaf extract of these plants was used as a reducing as well as stabilizing agent in the hydrothermal synthesis of silver nanoplates using silver nitrate as the precursor. Well dispersed silver nanoplates were formed. The appearance of two SPR (Surface Plasmon Resonance) bands corresponding to in-plane and out of plane vibration confirmed the formation of anisotropic nanostructures. The blue shift in peaks of the nanostructures in UV-visible spectra gave information about the stability of the nanoplates with time. Dynamic Light Scattering (DLS) and powder XRD were used to evaluate the ultimate average diameter and crystal structure of these nanostructures respectively. FESEM/EDX and HRTEM/SAED images also confirmed the formation of silver nanoplates. The FT-IR spectra helped to identify the reducing and stabilizing component of plant leaves extract in the formation of 2-D nanostructures. Preliminary antibacterial activity was examined using these nanoplates. A significant zone of inhibition was observed for S. aureus, a Gram positive bacterium.

Simultaneous determination of metoprolol succinate and amlodipine besylate in human plasma by liquid chromatography-tandem mass spectrometry method and its application in bioequivalence study.

A simple, sensitive and specific liquid chromatography-tandem mass spectrometry method was developed and validated for quantification of metoprolol succinate (MPS) and amlodipine besylate (AM) using hydrochlorothiazide (HCTZ) as IS in human plasma. Both the drugs were extracted by simple liquid-liquid extraction with chloroform. The chromatographic separation was performed on a reversed-phase peerless basic C18 column with a mobile phase of methanol-water containing 0.5% formic acid (8:2, v/v). The protonated analyte was quantitated in positive ionization by multiple reaction monitoring with a mass spectrometer. The method was validated over the concentration range of 1-100 ng/ml for MPS and 1-15 ng/ml AM in human plasma. The MRM transition of m/z 268.10-103.10, m/z 409.10-334.20 and m/z 296.00-205.10 were used to measure MPS, AM and HCTZ (IS), respectively. This method was successfully applied to the pharmacokinetic study of fixed dose combination (FDC) of MPS and AM formulation product after an oral administration to Indian healthy human volunteers.

Production and phenotypic analysis of rice transgenics with altered levels of pyruvate decarboxylase and alcohol dehydrogenase proteins.

Pyruvate decarboxylase (Pdc) and alcohol dehydrogenase (Adh) enzymes are responsible for the operation of ethanolic fermentation pathway that appears to correlate to an extent with anoxia tolerance in plants. This study was undertaken with the objective of (a) analysing the rice pdc gene family and (b) altering the efficacy of the ethanolic fermentation process, through production of transgenic rice plants over- and under-expressing pyruvate decarboxylase (employing Ospdc1 gene from rice) as well as over-expressing alcohol dehydrogenase (employing Ghadh2 gene from cotton) proteins. Correlations noted in this study between the pattern of expression of the Pdc alpha-subunit and Ospdc2 transcript as well as between the Pdc beta-subunit and Ospdc1 transcript suggest the possibility that alpha-subunit is encoded by Ospdc2 and that beta-subunit is encoded by Ospdc1. The fact that levels of Pdc beta-subunit were particularly high in pUH-sPdc1 (plasmid construct designed for over-expression of Ospdc1) seedlings while levels of beta-subunit levels were negligible or lower in pUH-asPdc1 (plasmid construct designed for under-expression of Ospdc1) seedlings also support these observations. Transgenics raised for over-expression of Pdc and Adh and under-expression of Pdc were confirmed for the transgene presence and effects by PCR, Southern blotting, Northern blotting, Western blotting and isozyme assays. Pdc and Adh over-expressing rice transgenics at early seedling stage under unstressed control growth conditions showed slight, consistent advantage in root vigour as compared to that of wild-type seedlings.

Isolation and transcription profiling of low-O2 stress-associated cDNA clones from the flooding-stress-tolerant FR13A rice genotype.

BACKGROUND: and Aims Flooding stress leads to a significant reduction in transcription and translation of genes involved in basal metabolism of plants. However, specific genes are noted to be up-regulated in this response. With the aim of isolating genes that might be specifically involved in flooding stress-tolerance mechanism(s), two subtractive cDNA libraries for the flooding-stress-tolerant rice genotype FR13A have been constructed, namely the single and double subtraction libraries (SSL and DSL, respectively). METHODS: To construct the SSL, mRNAs present in the unstressed control FR13A roots were subtracted from the mRNA pool present in low O2-stressed roots of FR13A rice seedlings. The DSL was constructed from mRNAs isolated from the roots of low O2-stressed FR13A rice seedlings from which pools of low-O2-stress up-regulated mRNAs from Pusa Basmati 1 and constitutively expressed mRNAs from FR13A roots were subtracted. RESULTS: In all, 400 and 606 cDNA clones were obtained from the SSL and DSL, respectively. Global transcript profiling by reverse northern analysis revealed that a large number of clones from these libraries were up-regulated by anaerobic stress. Importantly, selective up-regulated clones showed characteristic cultivar- and tissue-specific expression profiles. Sequencing and annotation of the up-regulated clones revealed that specific signal proteins, hexose transporters, ion channel transporters, RNA-binding proteins and transcription factor proteins possibly play important roles in the response of rice to flooding stress. Also a significant number of novel cDNA clones was noted in these libraries. CONCLUSIONS: It appears that cellular functions such as signalling, sugar and ion transport and transcript stability play an important role in conferring higher flooding tolerance in the FR13A rice type.

Molecular characterization of rice hsp101: complementation of yeast hsp104 mutation by disaggregation of protein granules and differential expression in indica and japonica rice types.

HSP100 protein is an important component of the heat-shock response in diverse organisms. Using specific primers based on cDNA sequence, rice hsp101 gene was PCR-amplified and sequenced. Southern analysis revealed that there appears to be a single gene per haploid genome coding for HSP101 protein in rice. Northern analysis showed that expression of hsp101 transcript is strictly heat-inducible and induction is transient in nature. In the temperature regime tested, 45 degrees C treatment to intact rice seedlings for 2 h showed maximal levels of hsp101 mRNA. Rice full-length hsp101 cDNA complemented yeast mutant disrupted for its own hsp104 gene by insertional mutagenesis, with efficacy that was comparable with Arabidopsis hsp101 cDNA. Electron micrographic evidence suggested that rice hsp101 cDNA in yeast is active in re-solubilizing the stress-induced protein granules in the post-stress recovery period. Rice hsp101 cDNA expression in hsp104 deficient yeast also caused recovery in tolerance against arsenite. Western analyses showed that this protein is expressed more rapidly during the stress period and retained for longer duration in the post-stress recovery period in japonica rice as compared to indica rice types. This is the first report wherein plant HSP100 protein expression is correlated to disappearance of protein granules in the yeast cells and distinct rice type-dependent protein expression patterns are reported.