Combined metabolome and transcriptome profiling provides new insights into diterpene biosynthesis in S. pomifera glandular trichomes

Background: Salvia diterpenes have been found to have health promoting properties. Among them, carnosic acid and carnosol, tanshinones and sclareol are well known for their cardiovascular, antitumor, antiinflammatory and antioxidant activities. However, many of these compounds are not available at a constant supply and developing biotechnological methods for their production could provide a sustainable alternative. The transcriptome of S. pomifera glandular trichomes was analysed aiming to identify genes that could be used in the engineering of synthetic microbial systems. Results: In the present study, a thorough metabolite analysis of S. pomifera leaves led to the isolation and structure elucidation of carnosic acid-family metabolites including one new natural product. These labdane diterpenes seem to be synthesized through miltiradiene and ferruginol. Transcriptomic analysis of the glandular trichomes from the S. pomifera leaves revealed two genes likely involved in miltiradiene synthesis. Their products were identified and the corresponding enzymes were characterized as copalyl diphosphate synthase (SpCDS) and miltiradiene synthase (SpMilS). In addition, several CYP-encoding transcripts were identified providing a valuable resource for the identification of the biosynthetic mechanism responsible for the production of carnosic acid-family metabolites in S. pomifera. Conclusions: Our work has uncovered the key enzymes involved in miltiradiene biosynthesis in S. pomifera leaf glandular trichomes. The transcriptomic dataset obtained provides a valuable tool for the identification of the CYPs involved in the synthesis of carnosic acid-family metabolites.General Secretariat of Research and Technology (GSRT) {[}09-SYN-23-879]; grant SEE-ERA. NET PLUS {[}ERA 64/01]; grant KRIPIS {[}MIS 448840

Inhibition of HIV-1 entry by extracts derived from traditional Chinese medicinal herbal plants

<p>Abstract</p> <p>Background</p> <p>Highly active anti-retroviral therapy (HAART) is the current HIV/AIDS treatment modality. Despite the fact that HAART is very effective in suppressing HIV-1 replication and reducing the mortality of HIV/AIDS patients, it has become increasingly clear that HAART does not offer an ultimate cure to HIV/AIDS. The high cost of the HAART regimen has impeded its delivery to over 90% of the HIV/AIDS population in the world. This reality has urgently called for the need to develop inexpensive alternative anti-HIV/AIDS therapy. This need has further manifested by recent clinical trial failures in anti-HIV-1 vaccines and microbicides. In the current study, we characterized a panel of extracts of traditional Chinese medicinal herbal plants for their activities against HIV-1 replication.</p> <p>Methods</p> <p>Crude and fractionated extracts were prepared from various parts of nine traditional Chinese medicinal herbal plants in Hainan Island, China. These extracts were first screened for their anti-HIV activity and cytotoxicity in human CD4+ Jurkat cells. Then, a single-round pseudotyped HIV-luciferase reporter virus system (HIV-Luc) was used to identify potential anti-HIV mechanisms of these extracts.</p> <p>Results</p> <p>Two extracts, one from <it>Euphorbiaceae</it>, <it>Trigonostema xyphophylloides </it>(TXE) and one from <it>Dipterocarpaceae</it>, <it>Vatica astrotricha </it>(VAD) inhibited HIV-1 replication and syncytia formation in CD4+ Jurkat cells, and had little adverse effects on host cell proliferation and survival. TXE and VAD did not show any direct inhibitory effects on the HIV-1 RT enzymatic activity. Treatment of these two extracts during the infection significantly blocked infection of the reporter virus. However, pre-treatment of the reporter virus with the extracts and treatment of the extracts post-infection had little effects on the infectivity or gene expression of the reporter virus.</p> <p>Conclusion</p> <p>These results demonstrate that TXE and VAD inhibit HIV-1 replication likely by blocking HIV-1 interaction with target cells, i.e., the interaction between gp120 and CD4/CCR5 or gp120 and CD4/CXCR4 and point to the potential of developing these two extracts to be HIV-1 entry inhibitors.</p

Ulvan, a bioactive marine sulphated polysaccharide as a key constituent of hybrid biomaterials: A review

Ulvan, a sulphated polysaccharide located in the cell walls of green algae that possesses unique structural properties albeit its repeating unit shares chemical affinity with glycosoaminoglycans, such as hyaluronan and chondroitin sulphate, has been increasingly studied over the years for applications in the pharmaceutical field. The increasing knowledge on ulvan&apos;s chemical properties and biological activities has triggered its utilization in hybrid materials, given its potential efficacy in biomedical applications. In the present review, the use of ulvan in the design of different biomaterials, including membranes, particles, hydrogels, 3D porous structures and nanofibers, is presented. The applications of these structures may vary from drug delivery to wound dressing or bone tissue engineering. In this context, general information regarding the structure and chemical variability, extraction processes, physicochemical properties, and biological activities of ulvan is reported. © 2019 Elsevier Lt

pH-Sensitive nanogates based on poly(l-histidine) for controlled drug release from mesoporous silica nanoparticles

The design and synthesis of novel poly(l-histidine)-grafted mesoporous silica nanoparticles (MSNs) by a surface-initiated ring-opening polymerization process (ROP) is reported. Using (3-aminopropyl)triethoxysilane (APTES) to introduce primary amino groups onto the MSN outer surface that work as ROP initiators, the nanoparticles were decorated with a uniform pH-sensitive poly(l-histidine) (PHis) shell. The method applied for the MSN functionalization, guaranteed that PHis chains were not grafted inside the MSNs&apos; nanochannels. Successful grafting of the PHis chains was confirmed by FT-IR spectroscopy, TEM and TGA, while the controlled character of the polymerization was monitored by SEC analysis. Dynamic light scattering (DLS) and zeta potential analysis were used to reveal the pH-responsive nature of the polypeptide-gated MSNs. The role of the grafted PHis chains as pH-sensitive nanogates for the MSN pores was verified by drug loading and release studies, using the model anticancer drug doxorubicin (DOX). DOX was efficiently loaded within the nanochannels of the hybrid MSN@PHis nanostructures (approximately 90%), and was released in a relatively controlled pH-triggered manner. Overall, the described materials are promising candidates as nanocarriers for potential drug delivery applications. © The Royal Society of Chemistry 2016

Development of multiple stimuli responsive magnetic polymer nanocontainers as efficient drug delivery systems

Magnetic nanodevices based on poly[(methacrylic acid)-co-(N- isopropylacrylamide)] [P(MAA-co-NIPAAm)] are prepared and used as drug delivery systems employing daunorubicin (DNR) as a model drug. The magnetic nanocontainers exploit the pH, temperature, and magnetic response of the polymeric shell constituents and magnetic nanoparticles, respectively, for controlled pH, temperature and alternating magnetic field triggered drug release. The in vitro cytotoxicity of both DNR-loaded and empty nanocontainers is examined on MCF-7 breast cancer cells along with the intracellular distribution of DNR. The results show that the DNR-loaded nanocontainers have an anti-tumor effect comparable to the free drug. The current observations provide important information for potent drug delivery and release systems. © 2014 WILEY-VCH Verlag GmbH &amp; Co. KGaA, Weinheim

Collagen from the marine sponges Axinella cannabina and Suberites carnosus: Isolation and morphological, biochemical, and biophysical characterization

In search of alternative and safer sources of collagen for biomedical applications, the marine demosponges Axinella cannabina and Suberites carnosus, collected from the Aegean and the Ionian Seas, respectively, were comparatively studied for their insoluble collagen, intercellular collagen, and spongin-like collagen content. The isolated collagenous materials were morphologically, physicochemically, and biophysically characterized. Using scanning electron microscopy and transmission electron microscopy the fibrous morphology of the isolated collagens was confirmed, whereas the amino acid analysis, in conjunction with infrared spectroscopy studies, verified the characteristic for the collagen amino acid profile and its secondary structure. Furthermore, the isoelectric point and thermal behavior were determined by titration and differential scanning calorimetry, in combination with circular dichroism spectroscopic studies, respectively. © 2017 by the authors