Response surface optimization of poly-ß-hydroxybutyrate synthesized by Bacillus cereus L17 using acetic acid as carbon source.

A strain of Bacillus that can tolerate 10 g/L acetic acid and use the volatile fatty acids produced by the hydrolysis and acidification of activated sludge to produce polyhydroxyalkanoate was screened from the activated sludge of propylene oxide saponification wastewater. The strain was identified by 16S rRNA sequencing and phylogenetic tree analysis and was named Bacillus cereus L17. Various characterization methods showed that the polymer synthesized by strain L17 is poly-ß-hydroxybutyrate, which has low crystallinity, good ductility and toughness, high thermal stability and a low polydispersity coefficient. It has wide thermoplastic material operating space as well as industrial and medicinal applications. The optimal fermentation conditions were determined by single factor optimization. Then, Plackett-Burman and Box-Behnken design experiments were carried out according to the single factor optimization results, and the response surface optimization was completed. The final results were: initial pH 6.7, temperature 25 °C, and loading volume 124 mL. The verification experiment showed that the yield of poly-ß-hydroxybutyrate after optimization increased by 35.2 % compared to that before optimization.

Deep 2-Hydroxyisobutyrylome in mouse liver expands the roles of lysine 2-hydroxyisobutyrylation pathway.

Lysine 2-hydroxyisobutyrylation (Khib), a newly characterized post-translational modification, is conserved in both eukaryotic and prokaryotic cells. At present, only about 6500 Khib sites have been identified in mammalian cells, which is insufficient compared with the well-known acetylation and thus hinders the understanding of its roles in diverse cellular processes. Here, utilizing immunoaffinity enrichment coupled with LC-MS/MS approach, we carried out a deep proteomics analysis of Khib in mouse liver. A total of 20861 Khib sites in 3768 proteins were identified, which expands the known Khib sites by two folds and represents the deepest Khib proteome in mammalian cells currently. Bioinformatics analysis showed that the 2-hydroxyisobutyrylated proteins have different subcellular localizations and participate in a wide range of molecular functions and cellular processes, such as metabolic processes and disease-related pathways. In addition, RNA-Seq analysis revealed that 1470 genes up-regulated and 790 genes down-regulated in response to elevated Khib levels in HeLa cells. The 1470 up-regulated genes were mainly associated with human papillomavirus infection, ECM-receptor interaction, as well as protein digestion and absorption, while the 790 down-regulated genes were mainly enriched in the multiple diseases and Glycolysis/Gluconeogenesis processes. Taken together, our research largely expands the known Khib sites, which helps delineate the biological functions of the Khib pathway and provides mechanistic insights into how Khib exerts its functions in specific cellular pathways.

Design, Synthesis, and Biological Evaluation of a Novel Series of Teriflunomide Derivatives as Potent Human Dihydroorotate Dehydrogenase Inhibitors for Malignancy Treatment.

Human dihydroorotate dehydrogenase (hDHODH), as the fourth and rate-limiting enzyme of the de novo pyrimidine synthesis pathway, is regarded as an attractive target for malignancy therapy. In the present study, a novel series of teriflunomide derivatives were designed, synthesized, and evaluated as hDHODH inhibitors. 13t was the optimal compound with promising enzymatic activity (IC50 = 16.0 nM), potent antiproliferative activity against human lymphoma Raji cells (IC50 = 7.7 nM), and excellent aqueous solubility (20.1 mg/mL). Mechanistically, 13t directly inhibited hDHODH and induced cell cycle S-phase arrest in Raji cells. The acute toxicity assay indicated a favorable safety profile of 13t. Notably, 13t displayed significant tumor growth inhibition activity with a tumor growth inhibition (TGI) rate of 81.4% at 30 mg/kg in a Raji xenograft model. Together, 13t is a promising inhibitor of hDHODH and a preclinical candidate for antitumor therapy, especially for lymphoma.

Controlled Delivery of Pan-PAD-Inhibitor Cl-Amidine Using Poly(3-Hydroxybutyrate) Microspheres.

This study deals with the process of optimization and synthesis of Poly(3-hydroxybutyrate) microspheres with encapsulated Cl-amidine. Cl-amidine is an inhibitor of peptidylarginine deiminases (PADs), a group of calcium-dependent enzymes, which play critical roles in a number of pathologies, including autoimmune and neurodegenerative diseases, as well as cancer. While Cl-amidine application has been assessed in a number of in vitro and in vivo models; methods of controlled release delivery remain to be investigated. P(3HB) microspheres have proven to be an effective delivery system for several compounds applied in antimicrobial, wound healing, cancer, and cardiovascular and regenerative disease models. In the current study, P(3HB) microspheres with encapsulated Cl-amidine were produced in a size ranging from ~4-5 µm and characterized for surface morphology, porosity, hydrophobicity and protein adsorption, in comparison with empty P(3HB) microspheres. Cl-amidine encapsulation in P(3HB) microspheres was optimized, and these were found to be less hydrophobic, compared with the empty microspheres, and subsequently adsorbed a lower amount of protein on their surface. The release kinetics of Cl-amidine from the microspheres were assessed in vitro and expressed as a function of encapsulation efficiency. There was a burst release of ~50% Cl-amidine in the first 24 h and a zero order release from that point up to 16 days, at which time point ~93% of the drug had been released. As Cl-amidine has been associated with anti-cancer effects, the Cl-amidine encapsulated microspheres were assessed for the inhibition of vascular endothelial growth factor (VEGF) expression in the mammalian breast cancer cell line SK-BR-3, including in the presence of the anti-proliferative drug rapamycin. The cytotoxicity of the combinatorial effect of rapamycin with Cl-amidine encapsulated P(3HB) microspheres was found to be 3.5% more effective within a 24 h period. The cells treated with Cl-amidine encapsulated microspheres alone, were found to have 36.5% reduction in VEGF expression when compared with untreated SK-BR-3 cells. This indicates that controlled release of Cl-amidine from P(3HB) microspheres may be effective in anti-cancer treatment, including in synergy with chemotherapeutic agents. Using controlled drug-delivery of Cl-amidine encapsulated in Poly(3-hydroxybutyrate) microspheres may be a promising novel strategy for application in PAD-associated pathologies.

Aligned silk fibroin/poly-3-hydroxybutyrate nanofibrous scaffolds seeded with adipose-derived stem cells for tendon tissue engineering.

In this work we investigated tenogenic differentiation of adipose-derived mesenchymal stem cells (AdMSCs), which were seeded onto silk fibroin/poly-3-hydroxybutyrate (SF/P3HB) scaffolds with aligned topography, and high mechanical strength. The electrospinning process was optimized by using the response surface method (RSM) and SF/P3HB nanofibrous matrices with a total polymer concentration of 5% (SF: PHB = 3: 1), flow rate 1 mL/h, collector rotation speed 2000 rpm, applied voltage 14 kV, and collector distance 25 cm were obtained. The average fiber diameter was 699 ± 203 nm and 80% of the nanofibers were aligned within the ±15o range. SF reinforcement reduced the crystallinity of P3HB, and the elastic modulus was found to be 197.0 ± 7.7 MPa. The scaffolds showed bacteriostatic effect. A 21-day of cell culture study was performed with rat rAdMSCs in the absence and presence of tenogenic differentiation factor-5 (GDF-5). The results demonstrated that SF/P3HB scaffolds allow the cells to proliferate and differentiate to the tenocytes. However, no significant effect of GDF-5 on the differentiation of cells was observed. These findings indicated that our aligned SF/P3HB scaffolds have a significant potential to be used for tendon tissue engineering.

Osteogenic differentiation of rat bone mesenchymal stem cells cultured on poly (hydroxybutyrate-co-hydroxyvalerate), poly (ε-caprolactone) scaffolds.

Bioresorbable biomaterials can fill bone defects and act as temporary scaffold to recruit MSCs to stimulate their differentiation. Among the different bioresorbable polymers studied, this work focuses on poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) and poly(ε-caprolactone) (PCL). Were prepared blends of PHBV and PCL to obtain PHBV based biomaterials with good tenacity, important for bone tissue repair, associated with biocompatible properties of PCL. This study assesses the viability of Vero cells on scaffolds of PHBV, PCL, and their blends and the osteogenic differentiation of mesenchymal stem cells (MSCs). Materials were characterized in surface morphology, DSC and Impact Strength (IS). Vero cells and MSCs were assessed by MTT assay, cytochemical and SEM analysis. MSC osteogenic differentiation was evaluated through alizarin red staining and ALP activity. We found some roughness onto surface materials. DSC showed that the blends presented two distinct melting peaks, characteristic of immiscible blends. IS test confirmed that PHBV-PCL blends is an alternative for increase the tenacity of PHBV. MTT assay showed cells with high metabolic activities on extract toxicity test, but with low activity in the direct contact test. SEM analysis showed spreading cells with irregular and flattened morphology on different substrates. Cytochemical study revealed that MSCs maintained their morphology, although in smaller number for MSCs. The development of nodules of mineralized organic matrix in MSC cultures was identified by alizarin red staining and osteogenic differentiation was confirmed by the quantification of ALP activity. Thus, our scaffolds did not interfere on viability of Vero cells or the osteogenic differentiation of MSCs.

Effects of Vanadyl Complexes with Acetylacetonate Derivatives on Non-Tumor and Tumor Cell Lines.

Vanadium has a good therapeutic potential, as several biological effects, but few side effects, have been demonstrated. Evidence suggests that vanadium compounds could represent a new class of non-platinum, metal antitumor agents. In the present study, we aimed to characterize the antiproliferative activities of fluorescent vanadyl complexes with acetylacetonate derivates bearing asymmetric substitutions on the ß-dicarbonyl moiety on different cell lines. The effects of fluorescent vanadyl complexes on proliferation and cell cycle modulation in different cell lines were detected by ATP content using the CellTiter-Glo Luminescent Assay and flow cytometry, respectively. Western blotting was performed to assess the modulation of mitogen-activated protein kinases (MAPKs) and relevant proteins. Confocal microscopy revealed that complexes were mainly localized in the cytoplasm, with a diffuse distribution, as in podocyte or a more aggregate conformation, as in the other cell lines. The effects of complexes on cell cycle were studied by cytofluorimetry and Western blot analysis, suggesting that the inhibition of proliferation could be correlated with a block in the G2/M phase of cell cycle and an increase in cdc2 phosphorylation. Complexes modulated mitogen-activated protein kinases (MAPKs) activation in a cell-dependent manner, but MAPK modulation can only partly explain the antiproliferative activity of these complexes. All together our results demonstrate that antiproliferative effects mediated by these compounds are cell type-dependent and involve the cdc2 and MAPKs pathway.

O-Heterocycle Synthesis via Intramolecular C-H Alkoxylation Catalyzed by Iron Acetylacetonate.

Intramolecular alkoxylation of C-H bonds can rapidly introduce structural and functional group complexities into seemingly simple or inert precursors. The transformation is particularly important due to the ubiquitous presence of tetrahydrofuran (THF) motifs as fundamental building blocks in a wide range of pharmaceuticals, agrochemicals, and natural products. Despite the various synthetic methodologies known for generating functionalized THFs, most show limited functional group tolerance and lack demonstration for the preparation of spiro or fused bi- and tricyclic ether units prevalent in molecules for pharmacological purposes. Herein we report an intramolecular C-H alkoxylation to furnish oxacycles from easily prepared α-diazo-ß-ketoesters using commercially available iron acetylacetonate (Fe(acac)2) as a catalyst. The reaction is proposed to proceed through the formation of a vinylic carboradical arising from N2 extrusion, which mediates a proximal H-atom abstraction followed by a rapid C-O bond forming radical recombination step. The radical mechanism is probed using an isotopic labeling study (vinyl C-D incorporation), ring opening of a radical clock substrate, and Hammett analysis and is further corroborated by density functional theory (DFT) calculations. Heightened reactivity is observed for electron-rich C-H bonds (tertiary, ethereal), while greater catalyst loadings or elevated reaction temperatures are required to fully convert substrates with benzylic, secondary, and primary C-H bonds. The transformation is highly functional group tolerant and operates under mild reaction conditions to provide rapid access to complex structures such as spiro and fused bi-/tricyclic O-heterocycles from readily available precursors.

Molecular recognition of tak-285 and lapatinib by inactive, active, and middle active-inactive HER2.

Experimental and theoretical studies have provided structural information regarding the shift from inactive to active EGFR, throughout which both conformations are linked via binding to specific tyrosine kinase inhibitors. For HER2, an intermediate active-inactive receptor conformation is present in the PDB, which has been co-crystallized with tak-285. The affinity of HER2 in monomeric state to tak-285 has been previously reported. However, the lack of structural knowledge of HER2 limits our capacity to understand whether tak-285, or other known HER2 inhibitors, selectively bind active, inactive, or intermediate forms of HER2. To elucidate mechanisms by which tak-285 binds to HER2, we first obtained information regarding the structural features of the active state of HER2 via microsecond MD simulations from the crystallized intermediate structure previously determined. Based on these HER2 conformers, together with the inactive HER2 conformer obtained in a previous study, we used docking and MD simulations coupled to MMGBSA approach to assess binding of tak-285 and lapatinib, known HER2/EGFR dual inhibitors, to HER2. Structural and energetic studies revealed that tak-285 binds with a greater affinity than lapatinib to active and intermediate active-inactive forms of HER2. This is in accordance with experimental findings that showed the tak-285 inhibitor has increased activity relative to lapatinib in breast cancer cell lines.

Fabrication and Characterization of the Core-Shell Structure of Poly(3-Hydroxybutyrate-4-Hydroxybutyrate) Nanofiber Scaffolds.

Tissue engineering scaffolds with nanofibrous structures provide positive support for cell proliferation and differentiation in biomedical fields. These scaffolds are widely used for defective tissue repair and drug delivery. However, the degradation performance and mechanical properties of scaffolds are often unsatisfactory. Here, we successfully prepared a novel poly(3-hydroxybutyrate-4-hydroxybutyrate)/polypyrrole (P34HB-PPy) core-shell nanofiber structure scaffold with electrospinning and in situ surface polymerization technology. The obtained composite scaffold showed good mechanical properties, hydrophilicity, and thermal stability based on the universal material testing machine, contact angle measuring system, thermogravimetric analyzer, and other methods. The results of the in vitro bone marrow-derived mesenchymal stem cells (BMSCs) culture showed that the P34HB-PPy composite scaffold effectively mimicked the extracellular matrix (ECM) and exhibited good cell retention and proliferative capacity. More importantly, P34HB is a controllable degradable polyester material, and its degradation product 3-hydroxybutyric acid (3-HB) is an energy metabolite that can promote cell growth and proliferation. These results strongly support the application potential of P34HB-PPy composite scaffolds in biomedical fields, such as tissue engineering and soft tissue repair.

Nonsolvent-induced phase separation of poly(3-hydroxybutyrate) and poly(hydroxybutyrate-co-hydroxyvalerate) blend as a facile platform to fabricate versatile nanofiber gels: Aero-, hydro-, and oleogels.

We demonstrated a strategy to prepare different types of 3-D nanofibrous polymeric gels, including hydro-, aero-, and oleogels by nonsolvent-induced phase separation (NIPS). NIPS-derived gel monoliths of poly(3-hydroxybutyrate) (PHB) and poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) blends were converted into hydrogels and aerogels by solvent exchange and freeze-drying, respectively. The high hydrophobicity and porosity of the nanofibrous PHB/PHBV aerogels enabled them to absorb various oils and swell to 20-30 times their own weight. The pseudo-second-order model was successfully used to describe the oil absorption behavior, and the obtained absorption rate constant increased with increasing PHBV content. The oil-swollen aerogels were highly elastic, thereby indicating that NIPS-derived aerogels are an excellent template for the fabrication of oleogels. With an increase in the PHBV ratio, the gels exhibited reduced modulus and collapse strength but increased collapse strain, thereby revealing higher ductility by compression. The rapid separation and re-binding of the liquid phase entrapped in the nanofiber network resulted in the unique thixotropic properties of the hydro- and oleogels. Indomethacin, a hydrophobic model drug, was successfully incorporated into injectable self-healing oleogels containing soybean oil and aerogels. These gels exhibited excellent cytocompatibility, and a better sustained drug release was observed for the oleogels compared to the aerogels.

Effect of the Intramolecular Hydrogen Bond on the Active Metabolite Analogs of Leflunomide for Blocking the Plasmodium falciparum Dihydroorotate Dehydrogenase Enzyme: QTAIM, NBO, and Docking Study.

BACKGROUND: Leflunomide (LFM) and its active metabolite, teriflunomide (TFM), have drawn a lot of attention for their anticancer activities, treatment of rheumatoid arthritis and malaria due to their capability to inhibit dihydroorotate dehydrogenase (DHODH) and Plasmodium falciparum dihydroorotate dehydrogenase (PfDHODH) enzyme. In this investigation, the strength of intramolecular hydrogen bond (IHB) in five analogs of TFM (ATFM) was analyzed employing density functional theory (DFT) using B3LYP/6-311++G (d, p) level and molecular orbital analysis in the gas phase and water solution. A detailed electronic structure study was performed using the quantum theory of atoms in molecules (QTAIM) and the hydrogen bond energies (EHB) of stable conformer obtained in the range of 76-97 kJ/mol, as a medium hydrogen bond. The effect of substitution on the IHB nature was studied by natural bond orbital analysis (NBO). 1H NMR calculations showed an upward trend in the proton chemical shift of the enolic proton in the chelated ring (14.5 to 15.7ppm) by increasing the IHB strength. All the calculations confirmed the strongest IHB in 5-F-ATFM and the weakest IHB in 2-FATFM. Molecular orbital analysis, including the HOMO-LUMO gap and chemical hardness, was performed to compare the reactivity of inhibitors. Finally, molecular docking analysis was carried out to identify the potency of inhibition of these compounds against PfDHODH enzyme. TFM acts as an inhibitor of dihydroorotate dehydrogenase (DHODH) and Plasmodium falciparum dihydroorotate dehydrogenase (PfDHODH) enzyme. Leflunomide and its active metabolite teriflunomide have been identified as drugs for treatment of some diseases, such as multiple sclerosis (MS), rheumatoid arthritis (RA), malaria, and cancer. Hydrogen bonds play a key role in the interaction between drugs and enzymes. OBJECTIVES: The aim of the present work is to investigate the effect of the strength of intramolecular hydrogen bonds (IHBs) in the active metabolite analogs of leflunomide or analogs of teriflunomide (ATFMs) and study the interaction of these inhibitors against the PfDHODH enzyme using quantum mechanical methods. METHODS: At first, intramolecular hydrogen bonds in five ATFMs were evaluated by the DFT method, quantum theory of atoms in molecules (QTAIM), nuclear magnetic resonance (NMR), natural bond orbital (NBO), and molecular orbital (MO) analyses. Then, the interaction of these inhibitors against the PfDHODH enzyme were compared using molecular docking study. RESULTS: All the computed results confirm the following trend in the intramolecular hydrogen bond strength in five mono-halo-substituted 2-cyano-3-hydroxy-N-phenylbut-2-enamide (ATFM): 5-FATFM> 4-Br-ATFM ≈ 3-Br-ATFM>3-Cl-ATFM>TFM-Z>2-F-ATFM which is in agreement with QTAIM, NMR, and NBO results. Docking results show that 5-F-ATFM (EHB=97kJ/mol) has the minimum MolDock score due to its considerable IHB strength. CONCLUSION: For strong IHBs (EHB>100kJ/mol), C=O and O-H group are involved in the intramolecular interactions and do not contribute to the external interactions. Also, the docking study revealed maximum binding energy between TFM-Z and PfDHODH enzyme.

Electrospun poly(3-hydroxybutyrate-co-4-hydroxybutyrate) /Octacalcium phosphate Nanofibrous membranes for effective guided bone regeneration.

Membranes used in guided bone regeneration (GBR) are required to exhibit high mechanical strength, biocompatibility, biodegradation, osteogenic and osteoinductive potential. In our study, poly(3-hydroxybutyrate-co-4-hydroxybutyrate)(P(3HB-co-4HB))/octacalcium phosphate (OCP) (P(3HB-co-4HB)/OCP) nanofibrous membranes were fabricated by electrospinning with two different P(3HB-co-4HB) to OCP ratios (P(3HB-co-4HB):OCP = 95:5 wt% and 90:10 wt%, termed P(3HB-co-4HB)/OCP(5)and P(3HB-co-4HB)/OCP (10), respectively) for GBR. The developed P(3HB-co-4HB)/OCP nanofibrous membranes were analysed for their osteogenic and osteoinductive properties using mesenchymal stem cells (MSCs) in vitro and in a calvarial bone defect rat model. The composite P(3HB-co-4HB)/OCP nanofibrous membranes showed decreased fibre size and enhanced tensile strength compared with those of P(3HB-co-4HB) nanofibrous membranes. In the in vitro studies, the P(3HB-co-4HB)/OCP membranes facilitated cell growth and osteoblastic differentiation of MSCs and were superior to P(3HB-co-4HB) membranes. After covered on the calvarial bone defects, P(3HB-co-4HB)/OCP membranes facilitated greater neobone formation than P(3HB-co-4HB) membranes did, as the result of histological evaluation and micro-CT analysis with higher bone volume/total volume (BV/TV) ratio and bone mineral density (BMD). P(3HB-co-4HB)/OCP(10) membranes with higher OCP content showed greater stiffness and osteoinductivity than P(3HB-co-4HB)/OCP (5)membranes, demonstrating the role of OCP in the composite membranes. These results indicated that electrospun P(3HB-co-4HB)/OCP nanofibrous membranes hold promise for the clinical application of GBR.

Hybrid nanobiocomposites based on poly(3-hydroxybutyrate) - characterization, thermal and mechanical properties.

Poly(3-hydroxybutyrate) is a biopolymer used to production of implants in the human body. On the other hand, the physical and mechanical properties of poly(3-hydroxybutyrate) are compared to the properties of isotactic polypropylene what makes poly(3-hydroxybutyrate) possible substitute for polypropylene. Unfortunately, the melting point of poly(3-hydroxybutyrate) is almost equal to its degradation temperature what gives very narrow window of its processing conditions. Therefore, numerous attempts are being made to improve the poly(3-hydroxybutyrate) properties. In the present work, hybrid nanobiocomposites based on poly(3-hydroxybutyrate) as a matrix with the use of organic nanoclay - Cloisite 30B and linear polyurethane as a second filler have been manufactured. The linear polyurethane was based on diphenylmethane 4,4'-diisocyanate and diol with imidazoquinazoline rings. The obtained nanobiocomposites were characterized by X-ray diffraction, scanning and transmission electron microscopies, thermogravimetry, differential scanning calorimetry and their selected mechanical properties were tested. The resulting hybrid nanobiocomposites have intercalated/exfoliated structure. The nanobiocomposites are characterized by a higher thermal stability and a wider range of processing temperatures compared to the unfilled matrix. The plasticizing influence of nanofillers was also observed. In addition, the mechanical properties of the discussed nanobiocomposites were examined and compared to those of the unfilled poly(3-hydroxybutyrate). The new-obtained nanobiocomposites based on poly(3-hydroxybutyrate) containing 1% Cloisite 30B and 5 wt. % of the linear of polyurethane characterized the highest improvement of processing conditions. They have the biggest difference between the temperature of degradation and the onset melting temperature, about 100 °C.

Usage of the newly synthesized poly(3-hydroxy butyrate)-b-poly(vinyl benzyl xanthate) block copolymer for vortex-assisted solid-phase microextraction of cobalt (II) and nickel (II) in canned foodstuffs.

The current research article was reported the synthesis of a novel poly(3-hydroxy butyrate)-b-poly(vinyl benzyl xanthate) block copolymer (PHB-Xa) for vortex-assisted solid-phase microextraction of cobalt(II) and nickel(II) from canned foodstuffs prior to their determinations by flame atomic absorption spectrometry. The block copolymer was synthesized and characterized by nuclear magnetic resonance spectroscopy and Fourier transform infrared spectroscopy. Experimental variables affecting the extraction efficiency of the copolymer were optimized. Since the PHB-Xa block copolymers have a high π conjugate structure and hydrophobicity, the use of this adsorbent yielded quantitative results for the extraction of Ni(II) and Co(II). After optimization, the linearities for Ni(II) and Co(II) were 0.05-80 ng mL-1 and 0.2-100 ng mL-1, respectively. The limits of detection and the limits of quantification were in the range of 0.015-0.06 ng mL-1 and 0.05-0.2 ng mL-1, respectively. The method was successfully applied to determination of Ni(II) and Co(II) in canned foodstuffs prepared by microwave digestion.

Characterization of Gemcitabine Loaded Polyhydroxybutyrate Coated Magnetic Nanoparticles for Targeted Drug Delivery.

BACKGROUND: Targeted drug delivery is one of the recent hot topics in cancer therapy. Because of having a targeting potential under the magnetic field and a suitable surface for the attachment of different therapeutic moieties, magnetic nanoparticles are widely studied for their applications in medicine. OBJECTIVE: Gemcitabine loaded polyhydroxybutyrate coated magnetic nanoparticles (Gem-PHB-MNPs) were synthesized and characterized for the treatment of breast cancer by the targeted drug delivery method. METHODS: The characterization of nanoparticles was confirmed by FTIR, XPS, TEM, and spectrophotometric analyses. The cytotoxicities of drug-free nanoparticles and Gemcitabine loaded nanoparticles were determined with cell proliferation assay using SKBR-3 and MCF-7 breast cancer cell lines. RESULTS: The release of Gemcitabine from PHB-MNPs indicated a pH-dependent pattern, which is a desirable release characteristic, since the pH of the tumor microenvironment and endosomal structures are acidic, while bloodstream and healthy-tissues are neutral. Drug-free PHB-MNPs were not cytotoxic to the SKBR-3 and MCF- 7 cells, whereas the Gemcitabine loaded PHB-MNPs was about two-fold as cytotoxic with respect to free Gemcitabine. In vitro targeting ability of PHB-MNPs was shown under the magnetic field. CONCLUSION: Considering these facts, we may suggest that these nanoparticles can be a promising candidate for the development of a novel targeted drug delivery system for breast cancer.

Bioevaluation of superparamagnetic iron oxide nanoparticles (SPIONs) functionalized with dihexadecyl phosphate (DHP).

Superparamagnetic iron oxide nanoparticles (SPIONs) have been investigated for wide variety of applications. Their unique properties render them highly applicable as MRI contrast agents, in magnetic hyperthermia or targeted drug delivery. SPIONs surface properties affect a whole array of parameters such as: solubility, toxicity, stability, biodistribution etc. Therefore, progress in the field of SPIONs surface functionalization is crucial for further development of therapeutic or diagnostic agents. In this study, SPIONs were synthesized by thermal decomposition of iron (III) acetylacetonate Fe(acac)3 and functionalized with dihexadecyl phosphate (DHP) via phase transfer. Bioactivity of the SPION-DHP was assessed on SW1353 and TCam-2 cancer derived cell lines. The following test were conducted: cytotoxicity and proliferation assay, reactive oxygen species (ROS) assay, SPIONs uptake (via Iron Staining and ICP-MS), expression analysis of the following genes: alkaline phosphatase (ALPL); ferritin light chain (FTL); serine/threonine protein phosphatase 2A (PP2A); protein tyrosine phosphatase non-receptor type 11 (PTPN11); transferrin receptor 1 (TFRC) via RT-qPCR. SPION-DHP nanoparticles were successfully obtained and did not reveal significant cytotoxicity in the range of tested concentrations. ROS generation was elevated, however not correlated with the concentrations. Gene expression profile was slightly altered only in SW1353 cells.

Modulation of neuronal cell affinity of composite scaffolds based on polyhydroxyalkanoates and bioactive glasses.

The biocompatibility and neuron regenerating properties of various bioactive glass (BG)/polyhydroxyalkanoate (PHA) blend composites were assessed in order to study their suitability for peripheral nerve tissue applications, specifically as lumen structures for nerve guidance conduits. BG/PHA blend composites were fabricated using Bioactive glass® 45 S5 (BG1) and BG 1393 (BG2) with the 25:75 poly(3-hydroxyoctanoate/poly3-hydroxybutyrate), 25:75 P(3HO)/P(3HB) blend (PHA blend). Various concentrations of each BG (0.5 wt%, 1.0 wt% and 2.5 wt%) were used to determine the effect of BG on neuronal growth and differentiation, in single culture using NG108-15 neuronal cells and in a co-culture along with RN22 Schwann cells. NG108-15 cells exhibited good growth and differentiation on all the PHA blend composites showing that both BGs have good biocompatibility at 0.5 wt%, 1.0 wt% and 2.5 wt% within the PHA blend. The Young's modulus values displayed by all the PHA blend/BG composites ranged from 385.6 MPa to 1792.6 MPa, which are able to provide the required support and protective effect for the regeneration of peripheral nerves. More specifically, the tensile strength obtained in the PHA blend/BG1 (1.0 wt%) (10.0 ± 0.6 MPa) was found to be similar to that of the rabbit peroneal nerve. This composite also exhibited the best biological performance in supporting growth and neuronal differentiation among all the substrates. The neurite extension on this composite was found to be remarkable with the neurites forming a complex connection network.

Characterization of PLA-P3,4HB active film incorporated with essential oil: Application in peach preservation.

This study aimed to develop an active film by using biodegradable materials and antioxidant essential oils to improve gas and water vapor permeability during peach preservation. O2 and CO2 volume fractions and water status were investigated by using an oxygen meter and conducting low-field nuclear magnetic resonance (LF-NMR), respectively. Results revealed that the film added with angelica essential oil (AEO) had a 49.4% increase in 2,2-diphenyl-1-picrylhydrazyl free radical scavenging activity and high O2 and CO2 transmission rates and water vapor permeability. The film added with AEO showed the best preservation effect, effectively delaying the oxidation of peach, maintained the combined water, and extended the shelf life of peaches to more than 15 days. This study provided a relatively new LF-NMR method for tracking the internal water status of packaged peaches and served as an effective reference for the development of active food packaging.