The rapid increase in the number of diabetic patients globally and exploration of alternate insulin delivery methods such as inhalation or oral route that rely on higher doses, is bound to escalate the demand for recombinant insulin in near future. Current manufacturing technologies would be unable to meet the growing demand of affordable insulin due to limitation in production capacity and high production cost. Manufacturing of therapeutic recombinant proteins require an appropriate host organism with efficient machinery for posttranslational modifications and protein refolding. Recombinant human insulin has been produced predominantly using E. coli and Saccharomyces cerevisiae for therapeutic use in human. We would focus in this review, on various approaches that can be exploited to increase the production of a biologically active insulin and its analogues in E. coli and yeast. Transgenic plants are also very attractive expression system, which can be exploited to produce insulin in large quantities for therapeutic use in human. Plant-based expression system hold tremendous potential for high-capacity production of insulin in very cost-effective manner. Very high level of expression of biologically active proinsulin in seeds or leaves with long-term stability, offers a low-cost technology for both injectable as well as oral delivery of proinsulin.

Chitosan is a deacetylated chitin which is found naturally, particularly in fungal cell walls and crustacean shells. Chitosan is biocompatible and fully biodegradable and is extensively analyzed for antimicrobial property. Chitosan has also been explored as a drug carrier due to its biocompatible properties. Some studies have demonstrated that use of chitosan to coat nanoparticles made of other materials would help in reducing their impact on the body and also increase their bioavailability. The molecular weight of chitosan and the degree of deacetylation can be modified to derive different physicomechanical properties. Chitosan exhibits potent antifungal activity against several fungal strains including Rhizopus oryzae, Aspergillus niger and Alternaria alternata. Various factors such as molecular weight, dose and functional groups attached to the chitosan have been shown to modulate the antifungal activity of chitosan. Chitosan is known to exert antifungal activity without the need for any chemical modification, however, new derivatives of chitosan can be created to target specific microbial pathogens. The development of novel and ecofriendly methods of chitosan nanoparticles (CSNPs) preparation is in progress for developing chitosan as an efficient antimicrobial agent and in drug delivery system. This review is to focus on recent application of CSNPs as antibacterial and antifungal agent and to highlight the effectiveness of employing chitosan with silver and other metal nanoparticles.

Rhazya stricta is a unique medicinal plant source for many indole alkaloids, non-alkaloids, flavonoids, triterpenes and other unknown molecules with tremendous potential for therapeutic applications against many diseases. In the present article, we generated computational data on predictive properties and activity across two key therapeutic areas of cancer and obesity, and corresponding cheminformatics studies were carried out to examine druggable properties of these alkaloids. Computed physiochemical properties of the 78 indole alkaloids from R. stricta plant using industry-standard scientific molecular modeling software and their predictive anti-cancer activities from reliable web-source technologies indicate their plausible therapeutic applications. Their predictive ADME properties are further indicative of their drug-like-ness. We believe that the top-ranked molecules with anti-cancer activity are clearly amenable to chemical modifications for creating potent, safe and efficacious compounds with the feasibility of generating new chemical entities after pre-clinical and clinical studies.

Increase in the incidence of Insulin Dependent Diabetes Mellitus (IDDM) among people from developed and developing countries has created a large global market for insulin. Moreover, exploration of new methods for insulin delivery including oral or inhalation route which require very high doses would further increase the demand of cost-effective recombinant insulin. Various bacterial and yeast strains have been optimized to overproduce important biopharmaceuticals. One of the approaches we have taken is the production of recombinant human insulin along with C-peptide in yeastPichia pastoris. We procured a cDNA clone of insulin from Origene Inc., USA. Insulin cDNA was PCR amplified and cloned into yeast vector pPICZ-α. Cloned insulin cDNA was confirmed by restriction analysis and DNA sequencing. pPICZ-α-insulin clone was transformed intoPichia pastoris SuperMan₅strain. Several Zeocin resistant clones were obtained and integration of insulin cDNA inPichiagenome was confirmed by PCR using insulin specific primers. Expression of insulin inPichiaclones was confirmed by ELISA, SDS-PAGE, and Western blot analysis. In vivo efficacy studies in streptozotocin induced diabetic mice confirmed the activity of recombinant insulin. In conclusion, a biologically active human proinsulin along with C-peptide was expressed at high level usingPichia pastorisexpression system.

The emergence of multidrug-resistant (MDR) microbes has become one of the major threat globally. Infectious diseases are the second leading cause of death, two-third of which are caused by Gram-negative bacteria. The increasing number of multidrug resistant (MDR) microbes is quite alarming and has raised the necessity of development of new antibacterial drugs. Escherichia coli and Klebsiella have been reported among the top most resistance-developing pathogens. Ricinus communis is an important medicinal plant reported to possess antimicrobial phytochemicals such as α-pinene. The hexane treated crude ethanolic extract of R. communis was evaluated against Gram-negative bacteria E. coli and Klebsiella oxytoca. The agar well diffusion assay was used to determine the antibacterial activity. In the present study, we have shown experimentally that leaf extract of R. communis can induce the deterioration of the inner and outer cell membranes of E. coli and K. oxytoca and decrease their viability at a concentration of 50 mg/ml. Transmission electron microscopic results revealed cell membrane damage, cellular disintegration and release of cytoplasmic content, leading to cell death. To our knowledge, this is the first study of the antibacterial activity of R. communis against E. coli and K. oxytoca by Transmission electron microscopy. The ultramicroscopic observations showed that the phytochemical present in the leaf extract of R. communis could penetrate the bacterial cell, causing rupture of cell membranes and hence confirm the cytotoxic and antimicrobial property of R. communis.

Ricinus communis is a traditional medicinal plant which has been utilized for centuries for treatment of various conditions. Due to the presence of diverse phytochemicals, Ricinus is an outstanding natural resource to discover new drugs for various diseases such as diabetes, cancer, arthritis, ulcer and asthma. In this study, we performed whole-genome gene expression profiling using RNA-Seq to determine the differentially expressed genes in a mammalian cell line after exposure to Ricinus leaf extract and elucidate their pharmacological effects in order to support its ethnomedicinal uses. Various genes involved in cancer, inflammation, atherosclerosis and diabetes were found to be differentially regulated after exposure to sub-lethal concentrations of the Ricinus extract in MCF7 cells. An important gene involved in cancer progression and metastasis, that is, PIK3R3 (Phosphatidylinositol 3-kinase regulatory subunit gamma), was downregulated in MCF7 cells after treatment with Ricinus extract. PIK3R3 is an important component of the PI3K/AKT signalling pathway which is essential for cell proliferation, angiogenesis, inhibition of apoptosis and metastasis to distant organs. The Ricinus extract downregulated the expression of DPP4 (Dipeptidyl peptidase-4) and upregulated the expression of PPAR-γ (Peroxisome proliferator-activated receptor gamma) which are crucial in controlling blood glucose levels. Expression of TNFAIP6 (Tumor necrosis factor-inducible gene 6), which is shown to mediate anti-inflammatory and protective effects, was increased after treatment with Ricinus extract. We also analyzed the genes which might also confer toxicity. Our gene expression profiling data corroborate the potential therapeutic benefits of Ricinus communis plant.