Chansu inhibits the expression of cortactin in colon cancer cell lines in vitro and in vivo

Background: Chansu is a transitional Chinese medicine that has been used for centuries as therapy for inflammation, anaesthesia and arrhythmia in China and other Asian countries. Recently, it has also been used for anti-cancer purposes. We have previously shown that Chansu has a huge pro-apoptotic potential on colon cancer cells, but to date the detailed mechanism of this action is not well understood. Methods: One of the major components of Chansu, Cinobufagin (CBF) was used to treat cancer cells. The expressions of levels of cortactin, an important factor in tumour progression and cancer invasion, were assessed in in vitro and in vivo experiments. Additional analyses were performed in subcellular protein fractions and immune-fluorescent staining was used to define cortactin protein expression and the changes of location in CBF-treated cells. Results: CBF strongly inhibited the expression of cortactin in HCT116 cells. There were reductions of both mRNA transcription and protein synthesis, which were more significant in the absence of oxygen in vitro. In addition, nuclear translocation of cortactin was observed in HCT116 cells post CBF exposure but not in the negative control, indicating that CBF is likely to interrupt co-localisation of cortactin to cytoskeletal proteins. Most importantly, CBF could diminish the expression of cortactin in human HCT116 xenograft tumours in nude mouse in vivo. Conclusions: CBF inhibits cortactin expression and nuclear translocation in colon cancer cells in vitro and in mouse models bearing human colon tumour in vivo, suggesting it might disrupt actin-regulated cell movement. Thus, CBF or Chansu could be developed as an effective anti-cancer therapy to stop local invasion and metastasis

The mechanisms of Chansu in inducing efficient apoptosis in colon cancer cells

Chansu is one of the most widely used traditional Chinese medicines in China, Japan, and other Southeast Asian countries primarily for antipain, anti-inflammation, and recently anticancer. Over 10 recipes and remedies contained Chansu, which are easily available in pharmacies and hospitals, but the mechanisms of action were not clearly articulated. In the present study, Cinobufagin (CBF), the major compound of Chansu, was employed as a surrogate marker to determine its ability in inducing cancer cell death. As expected, CBF has significant cancer-killing capacity for a range of cancers, but such ability differs markedly. Colon and prostate cancers are more sensitive than skin and lung cancers. Interestingly, cancer cells die through apoptotic pathway either being biphasic caspase- 3-dependent (HCT116) or independent (HT29). Multipathway analysis reveals that CBF-induced apoptosis is likely modulated by the hypoxia-inducing factor-1 alpha subunit (HIF-

Molecular characteristics of cement-lime treated contaminated-lateritic clay soil

The existence of contaminated land due to industrial and mining activities is an important apprehension in today's highly industrialized nations. However, the effect of these contaminants on the geotechnical properties of clayey soils can be modified by the chemical stabilization using traditional stabilizers like cement and lime, which result in achieving suitable material for construction purposes. Molecular characteristics of cement and lime stabilized contaminated- Lateritic clay soil presented in this research. This objective was applied by studying the induced alterations in molecular structure of cement-lime, and heavy metals stabilized lateritic soil using Fourier Transform Infrared (FTIR) spectroscopy. In current study, FTIR was performed to invistigate the molecular amendments in the structure of treated samples before and after contamination. Based on the results, the major vibrational bands were attributed to the kaolinite minerals. Furthermore, other bands appeared due to the presence of quartz and hematite. A new band was detected at 1462cm-1 with low intensity assigned to the carbonates, which was noticed increasing with time due to the producing of calcite. Also, it was detected a new sharp band at 1384cm-1 related to zinc and copper nitrates due the presence of heavy metals

Assessment of static and perfusion methods for decellularization of PCL membrane-supported periodontal ligament cell sheet constructs

Decellularization aims to harness the regenerative properties of native extracellular matrix. The objective of this study was to evaluate different methods of decellularization of periodontal ligament cell sheets whilst maintaining their structural and biological integrity.Human periodontal ligament cell sheets were placed onto melt electrospun polycaprolactone (PCL) membranes that reinforced the cell sheets during the various decellularization protocols. These cell sheet constructs (CSCs) were decellularized under static/perfusion conditions using a) 20 mM ammonium hydroxide (NH4OH)/Triton X-100, 0.5% v/v; and b) sodium dodecyl sulfate (SDS, 0.2% v/v), both +/- DNase besides Freeze-thaw (F/T) cycling method. CSCs were assessed using a collagen quantification assay, immunostaining and scanning electron microscopy. Residual fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) were assessed with Bio-plex assays.DNA removal without DNase was higher under static conditions. However, after DNase treatment, there were no differences between the different decellularization methods with virtually 100% DNA removal. DNA elimination in F/T was less efficient even after DNase treatment. Collagen content was preserved with all techniques, except with SDS treatment. Structural integrity was preserved after NH4OH/Triton X-100 and F/T treatment, while SDS altered the extracellular matrix structure. Growth factor amounts were reduced after decellularization with all methods, with the greatest reduction (to virtually undetectable amounts) following SDS treatment, while NH4OH/Triton X-100 and DNase treatment resulted in approximately 10% retention.This study showed that treatment with NH4OH/Triton X-100 and DNase solution was the most efficient method for DNA removal and the preservation of extracellular matrix integrity and growth factors retention

Polyisobutylene (PIB)-NHC Supported Catalysts for Cross-Coupling Reactions: A Green and Sustainable Protocol

N-Heterocyclic Carbenes (NHCs): Over the last two decades N-Heterocyclic carbenes (NHCs) have immensely attracted chemists in nearly all fields of chemistry. N-Heterocyclic carbenes are commonly encountered in coordination chemistry, they are extensively used as ligands for organometallic complexes. Perhaps the biggest hit of NHCs ligands was their use in Grubbs II catalyst for olefin metathesis chemistry. It is noteworthy that the success of NHCs ligands in catalysis is due to several factors favoring their high activity, selectivity and stability when compared to the phosphine counterparts in Grubbs I catalyst [1]. Supported Catalysts: Increased environmental and health awareness requires that designing new metal-catalysts should focus not only on increasing activity and selectivity but also on finding new strategies that help chemists recycle and separate the metal-catalyst from the reaction mixture. In general, homogenous catalysis is preferred over heterogeneous catalysis. This is due to the higher turnover number, better selectivity and usually lower operating temperatures required. On the other hand, heterogeneous catalysis has the advantage of the ease of separation of the catalyst from the final products and is generally less expensive. One important strategy is to use catalysts attached to a heterogeneous support and separate them from the products by simple filtration. Alternatively, homogeneous catalysts that can self-separate from the products by selective solvent extraction would be of great interest. The frequency of their reuse would be environmentally beneficial and to a higher extent this should overcome the lower activity of conventional heterogeneous catalysts. Metal catalysts that can self-separate from the reaction mixture are of great importance due to the reduced metal leaching into the product mixture. In addition, their reuse and recovery make this overall process much greener compared to the conventional homogeneous/heterogeneous catalysis systems. Ever since Herrmann et al. [2] reported the polystyrene supported NHC-palladium catalyst, studies have largely been focused on the use of polymeric supports for NHC-palladium catalysts. While polyethylene-glycol-supported catalyst can be extracted with a polar solvent, Bergbreiter et al. [3] and others have showed that polyisobutylene (PIB) is a useful support for ligands and their metal catalysts (Pd, Ru...) having preferable solubility towards solvents with low polarities such as hexanes, heptanes and decanes. In all of these biphasic systems for cross-coupling/olefin metathesis, the design is mainly focused on the recovery and the reuse of the supported catalysts. Biphasic catalysis having thermomorphic behavior have witnessed great developments due to their temperature-dependent miscibility [4]. While reactions in these biphasic mixtures can be conducted under homogeneous conditions at high-temperatures, the supported catalysts and the products/by-products can be efficiently separated by restoring the biphasic conditions at a low-temperature (Scheme 1). Herein we report the synthesis of new PIB-supported N-heterocyclic carbenes ligands having two different frameworks and their Pd-complexes, 1 and 2. The use, recovery and effectiveness of catalysts are detailed in both Heck and Suzuki cross-coupling reactions (Scheme 2). Metal leaching to the polar phase will be discussed too. Scheme 2: Heck cross-coupling and Suzuki cross-coupling using catalysts 1 and 2.qscienc

Secoisolariciresinol diglucoside protects against cadmium-induced oxidative stress-mediated renal toxicity in rats

Background: Cadmium is a well known environmental pollutant and strong toxic heavy metal, that causes oxidative damage to various organs of the body, including the kidney. Cadmium (II) chloride (CdCl2) is a water-soluble crystalline form, which exhibits a higher affinity with chlorides at the target site. The current study examined the protective effects of Secoisolariciresinol diglucoside (SDG), a principal lignan extracted from flaxseeds against CdCl2-induced renal toxicity in rats. Methods: Twenty four healthy male Wistar rats with four groups of six animals each were used in the study. Group-1- Control was administered with saline. Group-2 -was treated with SDG; Group-3 with CdCl2 alone, and Group-4 were treated with CdCl2 plus SDG. The effect of Cd on kidney was assessed in terms of various parameters like lipid peroxidation, production of Nitric oxide (NO) and Myeloperoxidase (MPO), and kidney function markers like uric acid, urea, and creatinine. The levels of antioxidant molecules like glutathione content and the activities of superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase were also measured, apart from histopathological studies. Results: The animals that received CdCl2, exhibited changes in the concentration of Cd in the kidney. The levels of kidney function markers like uric acid, urea, and creatinine were found to be abnormal in serum, and also there was a drastic decrease in the levels of glutathione content and the activities of superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase. The treatment of SDG significantly decreased (p < 0.05) the levels of NO and MPO in the animals treated with CdCl2 plus SDG when compared to the animal group treated with CdCl2 alone. The treatment of SDG before CdCl(2 )injection exhibited significant changes in the activity of the antioxidant enzymes, which was evidenced by the restoration in their activities, when compared to CdCl2 alone treated group (p < 0.05), as observed in the results of histopathology. Conclusions: The findings of the present investigation suggested that SDG exhibited anti-oxidant, anti-apoptotic and renoprotective properties. Thus, SDG may act as a supramolecular binding component and naturally occurring metal chelating agent for metal cations like Cd2+. Therefore, flaxseed lignan-SDG can be used as a therapeutic agent against nephrotoxicity caused by cadmium. However, detailed future studies are needed to know the underlying mechanism of action of SDG against the Cd and other heavy metals induced nephrotoxicity

Chemical Inhibitors and microRNAs (miRNA) Targeting the Mammalian Target of Rapamycin (mTOR) Pathway: Potential for Novel Anticancer Therapeutics

Abstract: The mammalian target of rapamycin (mTOR) is a critical regulator of many fundamental features in response to upstream cellular signals, such as growth factors, energy, stress and nutrients, controlling cell growth, proliferation and metabolism through two complexes, mTORC1 and mTORC2. Dysregulation of mTOR signalling often occurs in a variety of human malignant diseases making it a crucial and validated target in the treatment of cancer. Tumour cells have shown high susceptibility to mTOR inhibitors. Rapamycin and its derivatives (rapalogs) have been tested in clinical trials in several tumour types and found to be effective as anticancer agents in patients with advanced cancers. To block mTOR function, they form a complex with FKBP12 and then bind the FRB domain of mTOR. Furthermore, a new generation of mTOR inhibitors targeting ATP-binding in the catalytic site of mTOR showed potent and more selective inhibition. More recently, microRNAs (miRNA) have emerged as modulators of biological pathways that are essential in cancer initiation, development and progression. Evidence collected to date shows that miRNAs may function as tumour suppressors or oncogenes in several human neoplasms. The mTOR pathway is a promising target by miRNAs for anticance

Microbial sucrose isomerases: Producing organisms, genes and enzymes

Sucrose isomerase (SI) activity is used industrially for the conversion of sucrose into isomers, particularly isomaltulose or trehalulose, which have properties advantageous over sucrose for some food uses. All of the known microbial SIs are TIM barrel proteins that convert sucrose without need for any cofactors, with varying kinetics and product specificities. The current analysis was undertaken to bridge key gaps between the information in patents and scientific publications about the microbes and enzymes useful for sucrose isomer production.This analysis shows that microbial SIs can be considered in 5 structural classes with corresponding functional distinctions that broadly align with the taxonomic differences between producing organisms. The most widely used bacterial strain for industrial production of isomaltulose, widely referred to as " Protaminobacter rubrum" CBS 574.77, is identified as Serratia plymuthica. The strain producing the most structurally divergent SI, with a high product specificity for trehalulose, widely referred to as " Pseudomonas mesoacidophila" MX-45, is identified as Rhizobium sp.Each tested SI-producer is shown to have a single SI gene and enzyme, so the properties reported previously for the isolated proteins can reasonably be associated with the products of the genes subsequently cloned from the same isolates and SI classes. Some natural isolates with potent SI activity do not catabolize the isomer under usual production conditions. The results indicate that their industrial potential may be further enhanced by selection for variants that do not catabolize the sucrose substrate

Additively manufactured biphasic construct loaded with BMP-2 for vertical bone regeneration: A pilot study in rabbit

Vertical bone augmentation of the jaws is required when the height of bone is insufficient at the site of dental implant placement. In this proof of concept study, we investigated the potential of a biphasic polycaprolactone construct combined with a hyaluronic acid based hydrogel loaded with recombinant human bone morphogenetic growth factor-2 (BMP-2) for vertical bone regeneration. The biphasic scaffold consisted of an outer shell manufactured by fused deposition modelling, mimicking native cortical bone and providing mechanical and space maintenance properties essential for bone formation. Within this shell, a 90% porous melt electrospun microfibrous mesh mimicking the architecture of cancellous bone was incorporated in order to facilitate hydrogel loading and subsequent osteogenesis and angiogenesis. The in vitro performances of the biphasic construct demonstrated that BMP-2 was released in a sustained manner over several weeks and that cell viability was maintained in the hydrogel over 21 days. qRT-PCR demonstrated the upregulation of bone markers such as osteopontin, osteocalcin and collagen 1A1 at day 3 and 14 in the constructs loaded with BMP2. In vivo assessment of the biphasic scaffold was performed using a dose of 30 μg of BMP-2 in a rabbit calvarial vertical bone augmentation model. The histology and micro-CT analysis of the elevated space demonstrated that the hydrogel and the presence of BMP-2 enabled bone formation. However, this was limited to the immediate vicinity of the calvarial bone. The amount of newly formed bone was relatively small which was likely due to poor vascularisation of the extraskeletal space. The utilisation of this biomimetic biphasic construct with excellent space maintenance properties can be of interest in dentistry although the in vivo model requires refinement to demonstrated appropriate efficacy