Current Pharmaceutical Biotechnology - Volume 15, Issue 2, 2014

In this work, the stability of Bevacizumab (Avastin®) repackaged in individual 1 mL single-use syringes and stored at different conditions was assessed. Bevacizumab repackaged in single-use syringes results from the off-label use of the drug as an intravitreal agent in the treatment of retinal diseases. Bevacizumab stability was assessed by assaying the anti-VEGF activity using an indirect ELISA method and a Dynamic Light Scattering study. The thermal stability of the drug was also studied by calorimetric analysis, aimed to evaluate thermodynamic parameters associated to the thermal unfolding process. Furthermore, microbiological and fungal tests on the Bevacizumab syringes were performed. As a result, a significant decrease of the anti-VEGF activity was detected when syringes were exposed to UV light at a temperature of 37°C. Under these conditions, the Dynamic Light Scattering study showed an increase of the average size of Bevacizumab; probably due to aggregation. In conclusion, Bevacizumab stability, when stored under different conditions, was assessed considering three different aspects: anti-VEGF activity, microbial contamination and physico-chemical properties. Bevacizumab was found to be stable, under sterile conditions, for 3 months at 4°C and for 7 days at room temperature, exposed to indirect light sources, while a brief exposure of the drug to direct UV radiation proved detrimental to drug stability.

Platelet-rich plasma (PRP) is widely used to promote tissue repair and accelerate osteogenesis, but there is no agreement about its mechanism of action. We characterized the modulatory effect of PRP on the in vitro osteoblast model SaOS-2, by using cell motility/chemoattraction and osteogenesis/mineralization assays, and a series of osteogenic/ osteoclastogenic genomic markers. Scratch wound assay showed that PRP stimulates cell motility, while transwell assay revealed a strong chemoattraction. Alkaline phosphatase (ALP) and alizarin red-S assays showed that PRP induces slight, but significant, stimulations of ALP activity and mineralization. The TGF-β inhibitor SB431542 reversed these effects, showing a main role for TGF-β1 released by PRP. Analyses of gene expression by qRT-PCR, showed the upregulation of osteocalcin, osteopontin, osteoprotegerin, receptor activator of NFαB (RANK), and runt-related transcription factor 2 (RUNX2) genes, with a total reversion by SB431542 for osteoprotegerin and RANK, and a partial reversion for ostecalcin, osteopontin, and RUNX2. The use of PCR array technique revealed the upregulation of the cathepsin K gene. These data show that PRP induces the development of mixed osteogenic/osteoclastogenic traits in the SaOS-2 model. Such a behavior may favour in vivo bone resorption and reconstitution at post-surgery or post-traumatic sites.

Leukemia remains a major cause of death in both developing as well as developed nations worldwide. Development of novel therapeutic approaches is perceived to be the best strategy to combat this disease. Availability of limited effective drugs with least side effects is the major hurdle in treating leukemia. Another major concern remains initiation and engagement of therapeutic strategies that may subvert or overcome drug-resistance. Deployment of plant-based molecules in therapeutic engagements is regarded to be the most apt approach to combat different forms of leukemia. Diterpenoids of natural origin have a broad range of therapeutic activities including anti-leukemic effect. In vitro and in vivo studies establish many diterpenoids from natural sources and their synthetic derivatives to have appreciable cytotoxic and apoptotic efficacies against a series of leukemic cells. This paper attempts to identify and describe diterpenoids with their plant sources and analyze the molecular mechanistic approaches underlying their effective apoptotic activities against leukemic cells studied till date. The present review article summarizes different categories of diterpenoids having various degrees of anti-leukemic activities and discusses the measures of overcoming the bio-availability issues of these natural molecules so that they can serve as extremely viable options in future leukemia therapy.

As sessile and filter-feeding metazoans, marine sponges represent an ecologically important and highly diverse component of marine benthic communities throughout the world. It has been suggested that marine sponges are hosts to many microorganisms which can constitute up to 40-60% of its biomass. Recently, sponges have attracted a high interest from scientific community because two important factors. First there is the fact that sponges have a wide range of associated bacteria; and, second, they are a rich source of bioactive substances. Since 1950, a number of bioactive substances with various pharmacological functions have been isolated from marine sponges. However, many of these substances were subsequently shown to be actually synthesized by sponge-associated bacteria. Bacteria associated with marine sponges constitute an interesting source of novel bioactive compounds with biotechnological potential such as antimicrobial substances, enzymes and surfactants. In addition, these bacteria may be biofilm forming and can act as bioindicators in bioremediation processes of environmental pollution caused by oil and heavy metals. This review focuses on the biotechnological applications of these microorganisms.

Dengue infections produce a distinct character of virus-induced intracellular membrane alterations which are associated with the viral replication machinery. Currently, the NS3 protein is being targeted for antiviral therapy against dengue. NS3 protein of dengue virus interacts with nuclear receptor binding protein (NRBP) of human causing cell trafficking between the Endoplasmic Reticulum (ER) and Golgi, which interacts with Rac3, a member of the Rho-GTPase family. No crystal structure of the NRBP is available for any species, thus limiting the complete understanding of structure- function relationships of this protein. The present study deals with the molecular modeling of the viral protein (NS3 of DENV1-4), the host protein (NRBP) and their interactions through protein-protein docking study. Theoretical threedimensional structures of the NRBP and NS3 were modeled using the Modeller 9v8, and the evaluated models were docked using GRAMM-X to study the mode of protein-protein interaction (NRBP as receptor and NS3 as ligand). The docked docking complexes were further evaluated for interaction analysis by the RosettaDock Server. Suface and interface residues were observed along with hydrogen and hydrophobic interaction. The conserved residues forming hydrogen interaction of NRBP with DENV1-4 serotypes were found to be GLN 305, SER 363 and GLN 379.

α-(-)-bisabolol is a natural monocyclic sesquiterpene present in the essential oil has generated considerable interest in the chemical and pharmaceutical industries and currently in use in various formulations, mainly in cosmetics. This study was undertaken to evaluate its therapeutic profile against skin inflammation using in-vitro, in-vivo and in-silico assays. Lipopolysachharide (LPS) and 12-O-tetradecanoyl-phorbol-13-acetate (TPA)-induced production of proinflammatory cytokines (TNF-α and IL-6) in macrophage cells as well as in TPA-induced skin inflammation in mice was significantly inhibited by α-(-)-bisabolol. TPA-induced ear thickness, ear weight and lipid peroxidation and histopathological damage in the ear tissue were also significantly inhibited by topical application of α-(-)-bisabolol in a dose dependent manner. In-vitro and in-vivo toxicity profiles indicate that it is safe for topical application on skin. Molecular docking study also revealed its strong binding affinity to the active site of the pro-inflammatory proteins. These findings suggested that α-(-)-bisabolol may be a useful therapeutic candidate for the treatment of skin inflammation.

The goal of the research work entertained herein was the development and characterization of a poly-(vinyl alcohol) (PVA) hydrogel cross-linked with glutaraldehyde and impregnated with 0.2% (w/w) nitrofurazone (NTZ), for topical applications. To verify the active principle release capability, one has determined (i) swelling profile, (ii) in vitro release of NTZ via UV-VIS spectrophotometry, and (iii) antimicrobial activity via exposure to the hydrogel of ATCC strains of Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa. The optimized hydrogel was further characterized via scanning electron microscopy (SEM), infrared spectroscopy with Fourier transform, moisture content determinations and thermal analyses via thermal gravimetry (TGA). Swelling tests revealed a mass increase from 100±5% up to 350±11%. Incorporated NTZ displayed bactericidal activity, as expected, being released in a linearly controlled fashion above 6 μg/mL during experiment timeframes of 14 h. SEM analyses allowed verification of a homogeneous surface morphology, while infrared spectra showed that NTZ did not bind strongly to the cross-linked polymer. Furthermore, results from thermal analyses suggested a loss of thermal stability arising from incorporation of NTZ in the hydrogel. The optimized hydrogel exhibited characteristics with high potential for (antimicrobial) treatment of skin lesions.