Current Pharmaceutical Biotechnology - Volume 17, Issue 12, 2016

Stereospermum fimbriatum is one of the medicinal plants that has been claimed to be used traditionally to treat several illnesses such as stomachache, earache, skin irritation and postpartum illness. The genus of this plant is known to possess medicinal properties in every part of the plant. Therapeutic potential of S. fimbriatum is anticipated based on numerous previous studies that documented variety of phytochemical contents and bioactivity of the genus. The most reported bioactivities of its genus are antimicrobial, antioxidant, anti-diabetic, anti-inflammatory, anti-diarrheal and analgesic activities. S. fimbriatum is a rare species that has not been discovered yet. Thus, this review aims at highlighting the potentials of S. fimbriatum by collecting available data on the bioactivities of its genus and set the directions for future research on this plant.

Introduction: Development of new and improved mosquito control methods, that are economically sustainable and effective, is a critical requirement in the management of vector-borne epidemic diseases. Aedes albopictus is one of the main vectors of various important pathogens in the tropics, which now have the potential to also spread in temperate regions, owing to the environmental and climate changes in act. Materials and Methods: We report about the isolation of steroidal saponins from Dracaena arborea by fractionation followed by column separation. The obtained fractions and/or pure compounds were tested by biological essays for their insecticidal activity against A. albopictus larvae. Results: Various compounds were found to exert larvicidal effects. In specific, spiroconazole A demonstrated the best insecticidal activity, showing LT50 value of 57.23 hours at 25 ppm. Discussion: We finally discuss about the value of this finding in the context of the present strategies of Integrated Mosquito Management.

Toxicity assessment is an important tool in drug discovery and development. PT-31 (3-(2-chloro-6-fluorobenzyl)-imidazolidine-2,4-dione) is an imidazolidine- 2,4-dione analogue of clonidine that displays a dose-dependent analgesic profile and synergism with morphine. This study investigated genotoxic and mutagenic effects of PT-31 in Swiss mice. For this, ten mice (M1:F1) per group were treated with PT-31 intraperitoneally (i.p.) at 0.5, 1.0 and 5.0 mg/kg. The dimethyl sulfoxide (0.5%) and 50 mg/kg cyclophosphamide (i.p.) were taken as negative (NC) and positive controls, respectively. The bone marrow cells were collected after 24 h, while peripheral blood after 30 min, 12 h and 24 h of the treatment for the comet assay. Micronucleus (MN) test was performed only on bone marrow cells collected after 24 h of i.p. treated animals. A hundred cells were considered for the comet assay and quantification of the index of damage and frequency of damage. Lack of genotoxicity with 0.5 mg/kg of PT-31 and DNA repair ability with 0.5 and 1.0 mg/kg doses at 12 h and 24 h in comparison to NC group was observed (P<0.05). There was an increase in MN formation by PT-31 1.0 and 5.0 mg/kg treated female and male mice, respectively. PT-31 induced genotoxic and mutagenic effects only in higher doses.

Background: Multifunctional nanostructures have received great deal of attention in biomedical area due to their capabilities in the development of new therapeutic and diagnostic agents. Silver and iron oxide nanoparticles, owing to their specific characteristics, are considered to develop bifunctional hybrid nanoparticles for magnetic delivery of silver nanoparticles as cytotoxic agent toward cancer cells. Objective: This study was designed to explore the in-vitro cytotoxic and apoptotic activities of three different silver-iron oxide binary hybrid nanoparticles on human liver hepatocellular carcinoma cell line. Method: Three different silver-iron oxide binary hybrid nanoparticles were synthesized and characterized through the designed procedures. Apoptosis induction was investigated through flow cytometry and the influence on bax gene expression level was analyzed using quantitative real time polymerase chain reaction. Results: All the three types of silver-iron oxide hybrid nanoparticles (possessing different characteristics) exhibited cytotoxic and apoptotic effects. Furthermore, the up regulation of bax gene expression suggested the involvement of the intrinsic pathway of apoptosis. Some of the transcription regulators which could interact with bax gene promoter were analyzed and found out to be mostly contributed in the stress responses. Among the test nanoparticles, the strongest cytotoxic and apoptotic effect was induced by the binary hybrid nanoparticle which was synthesized with glucose as reducing agent; suggesting that the biological activity was affected by different characteristics of the designed nanoparticles. Conclusion: Combined properties of silver and magnetic nanoparticles in the binary hybrid nanoparticles, provide a great potential to be exploited in the cancer therapy, where the combination of cytotoxicity and magnetic targeting is desired.

We aim to determine the regulation of apoptosis by paclitaxel-induced and understand cancer dynamics to treatment targets for HeLa cells by identifying decrease/increase genes expression on HeLa cells. In this study, the anti-tumor effects of Paclitaxel (PAC) on HeLa cells have been studied in order to determine the cellular and molecular mechanisms of these effects. PAC has been applied to HeLa cells in 6 different doses (3, 7.5, 15, 30, 60, 120 nM) for 48 hours and the IC50 dose MTT method, has been determined with apoptic index (AI) DAPI. Morphological aspects have been demonstrated using light, phase contrast and fluorescent microscopes, additionally activation of Caspase 3,7 and 10 have been shown using florescent spectroscopy. RT-PCR and qRT-PCR have been used to evaluate pro/anti-apoptotic gene expression. According to the parameters being evaluated; PAC has reduced cell multiplication based on dosage and time (p<0.01). 15 nM has been determined as the IC50 value. AI value has been determined as 42%. In the molecular level analyses in addition to the increase in Caspase3,7,10 activation, RT-PCR results show that bax, bak, bcl-x, bik, mcl-1 genes are expressed in the control group as well as the experimental 15 nM group; whereas bak, bcl-x ve bik genes have a decrease in expression compared to the control group. qRT-PCR results show that Apaf1, Bad, Bax, Bcl2L11, Caspase1, Caspase10, Caspase4, Caspase7, Dffa, Fas, Htra2, Lrdd, NFKB1, NFKB2, PMAIP1, RELA, RELB, TNFRSF10A, TNFRSF10C, TNFRSF10D, TNFRSF1A, TNFRSF21, TNFRSF25 gene expressions have increased significantly. On the other hand, BAG1, BBC3, Bcl2L1, Bcl2L10, Bid, Caspase2, Caspase6, Caspase8, Caspase9, FADD, FAM96A, FasLG, HRK, SOCS3, TNF, TNFSF10, TRAF5, TRAF6 mRNA levels are significantly decreased.

Background: Ultraviolet irradiation is able to deeply penetrate into the dermis and alter fibroblast structure and function, leading to a degradation of the dermal extracellular matrix. Objectives: The regenerative effect of plasma rich in growth factors (PRGF) on skin ageing was investigated using UVB photo-stressed human dermal fibroblasts as an in vitro culture model. Method: PRGF was assessed over the main indicative features of ultraviolet B irradiation, including ROS formation, cell viability and death detection, apoptosis/ necrosis analysis and biosynthetic activity measurement. Four different UV irradiation protocols were tested in order to analyze the beneficial effects of PRGF. Results: Ultraviolet irradiation exhibited a dose dependent cytotoxicity and dose of 400mJ/cm2 was selected for subsequent experiments. PRGF increased the cell viability and decreased the cell death comparing to the non-treated group. The apoptosis and necrosis were significantly lower in PRGF treated fibroblasts. ROS production after UV irradiation was significantly reduced in the presence of PRGF. Procollagen type I, hyaluronic acid and TIMP-1 levels were higher in the when treated with PRGF. Conclusion: This preliminary in vitro study suggests that PRGF is able to prevent UVB derived photooxidative stress and to diminish the cell damage caused by ultraviolet irradiation.

Background: Regenerative strategies based on the use of platelet concentrates as an autologous source of growth factors (GF) has been proposed to promote the healing of long bone nonunions. However, the relatively high failure rate stimulates interest in growing knowledge and developing solutions to obtain the best results from the regenerative approach. Objective: In this study we evaluated whether a cell-based assay system could be able to recognize patients who will benefit or not from the use of autologous platelet preparations. Method: The autologous serum was used in culture medium to promote the osteogenic differentiation of normal bone-marrow stromal cells (BMSC). Blood samples were collected from 16 patients affected by aseptic long bone nonunion who were candidates to the treatment with autologous platelet-rich fibrin. The osteoinductive effect was detected by measuring the BMSC proliferation, the mineralization activity, and the expression of bone-related genes. Serum level of basic fibroblast growth factor (bFGF) was considered as a representative marker of the delivery of osteogenic GFs from platelets. Laboratory results were related to the characteristics of the disease before the treatment and to the outcome at 12 months. Results: Serum samples from "good responders" showed significantly higher levels of bFGF and were able to induce a significantly higher proliferation of BMSC, while no significant differences were observed in terms of osteoblast differentiation. Conclusion: BMSC-based assay could be a useful tool to recognize patients who have a low probability to benefit from the use of autologous platelet concentrate to promote the healing of long bone nonunion.

Glucose-1-Phosphate Thymidylyltransferase (RmlA) is one of the enzymes in rhamnose biosynthesis pathway, where rhamnose acts as linker of peptidoglycan and arabinogalacton in the cell wall, therefore RmlA is a potential enzyme for the survival of Mycobacterium tuberculosis (Mtb). To go into the depth of the structure for exploring binding regions, homology model of RmlA was built in Prime, Schrodinger v9.2. The model with lowest Discrete Optimized Potential Energy (DOPE) score of -35524.17 kcal/mol and RMSD of 0.1 Å with the template (1H5R_B) was subjected to Molecular Dynamics Simulation (MDS) for 5 ns to achieve its stable folding state. The tertiary structure of the proposed model is composed of α/β/α sandwich type protein with quasi-Rossmann type folding pattern. The substrate, deoxy Thymidine tri phosphate (dTTP) comprises of triphosphate (R1) and methyl (R2) side chains where, R1 is highly essential for the survival of Mtb. Therefore, nineteen side chain analogues of dTTP were designed by substituting R1 and R2 chain of dTTP using Combi Glide, Schrodinger v9.2 and docked with the target RmlA protein. Out of which two analogues such as, 6-[(2R,3S,5R)-5-[5-(2- aminoethyl)-2,4-dioxo-1,2,3,4-tetrahydropyrimidin-1-yl]-3-hydroxyoxolan-2 yl] hexanoic acid (COMP- 11) and 4-(2-{1-[(1S,3S,4S)-3-(5-carboxypentyl)-4-hydroxy-2-methylidenecyclopentyl]-2,4-dioxo- 1,2,3,4-tetrahydropyrimidin-5-yl}ethyl)morpholin-4-ium (COMP-12) showed the highest GLIDE score (-12.55 Kcal/mol and -11.58 Kcal/mol respectively) than that of substrate (-9.725 Kcal/mol). During simulations, hydrogen bonding profile between the two top hits and protein ranges up to 5 strong polar contacts which were much stronger than that of substrate. Similarly, the computational binding free energy of both the analogues was found to be less than -70 Kcal/mol which is much lower than that of substrate (-52.84 Kcal/mol). All these results suggest that these two compounds have more stable interaction than that of substrate inside the solvent condition and can be used as competitive inhibitors.

Breast cancer (BC) remains as one of the important causes of cancer deaths among women globally. Therefore, finding an effective treatment for BC is really needed. Cancer immunotherapy, as an emerging field, has a notable role in BC therapy. Peptide vaccines possess an outstanding role among different strategies in cancer immunotherapy. In vaccine design for cancer, induction of cellular and humoral immune responses should be considered. In the current study, cytolytic T lymphocytes (CTL) epitopes were evoked from human epidermal growth factor receptor (HER2), mucin 1 protein (MUC1), and heparanase antigenic proteins; and helper T lymphocytes (HTL) epitopes were determined from survivin protein by various immunoinformatics servers. Furthermore, our vaccine peptide contains several linear and conformational B cell epitopes that can induce humoral immunity. In order to elicit broad cellular and humoral immune responses, Por B protein from Neisseria meningitides, which is one of the toll like receptor 2 (TLR2) agonists, was utilized as an adjuvant in the vaccine construct. The designed peptide vaccine contains the extracellular domain of murine ULBP-like transcript 1 (MULT1), which binds to a natural killer group 2 member D (NKG2D) receptor with a high affinity and has a key role in triggering the innate immune response. All the mentioned segments were fused together by functional and structural amino acid linkers. Taken together, we project that our vaccine construct can potentially induce cellular, humoral, and innate immune responses in BC patients.