Biosynthesis of exopolysaccharide from waste molasses using Pantoea sp. BCCS 001 GH: a kinetic and optimization study.

The bacterium Pantoea sp. BCCS 001 GH produces an exopolysaccharide (EPS) named Pantoan through using sugar beet molasses (SBM) as an inexpensive and widely available carbon source. This study aims to investigate the kinetics and optimization of the Pantoan biosynthesis using Pantoea sp. BCCS 001 GH in submerged culture. During kinetics studies, the logistic model and Luedeking-Piret equation are precisely fit with the obtained experimental data. The response surface methodology (RSM)-central composite design (CCD) method is applied to evaluate the effects of four factors (SBM, peptone, Na2HPO4, and Triton X-100) on the concentration of Pantoan in batch culture of Pantoea sp. BCCS 001 GH. The experimental and predicted maximum Pantoan production yields are found 9.9 ± 0.5 and 10.30 g/L, respectively, and the best prediction factor concentrations are achieved at 31.5 g/L SBM, 2.73 g/L peptone, 3 g/L Na2HPO4, and 0.32 g/L Triton X-100 after 48 h of submerged culture fermentation, at 30 °C. The functional groups and major monosaccharides (glucose and galactose) of a purified Pantoan are described and confirmed by 1HNMR and FTIR. The produced Pantoan is also characterized by thermogravimetric analysis and the rheological properties of the biopolymer are investigated. The present work guides the design and optimization of the Pantoea sp. BCCS 001 GH culture media, to be fine-tuned and applied to invaluable EPS, which can be applicable in food and biotechnology applications.

The potential of surface nano-engineering in characteristics of cobalt-based nanoparticles and biointerface interaction with prokaryotic and human cells.

Cobalt-based nanoparticles (CBNPs) have recently received great attention in biomedical studies; however, the possible biotoxicity of these nanoparticles (NPs) has remained a foremost concern that should be addressed. As surface functionalization is one of the helpful proposed solutions, we aimed to apply Lipoamino acids (LAAs) as a coating agent to improve biocompatibility. To this purpose, cobalt oxide, cobalt ferrite, and iron oxide nanoparticles (IONs) were synthesized with and without 2-amino-hexadecanoic acid coating to assess the impacts of LAA coating on characteristics and biocompatibility of CBNPs in human cells and compare with IONs, a widely used magnetic NPs in biomedicine. Antibacterial activities of NPs were evaluated against four Gram-negative and Gram-positive bacteria species to assess their biointerface interaction with prokaryotic cells. In addition, the antibacterial activities of synthesized NPs were compared to silver NPs, one of the widely used antimicrobial NPs and standard antibiotics (ampicillin). The structural characteristics properties of NPs were analyzed using TEM, FE-SEM, EDS, FTIR, XRD, and VSM. These NPs exhibited sphere-like to polygon-like morphology with desirable mean size. CBNPs displayed dose-dependent cytotoxicity and antimicrobial activities against human cell lines and all tested microbial species, as well as more cytotoxicity and bacterial inhibition compared to IONs. Besides, the results revealed that LAA coating could significantly improve the biocompatibility and antibacterial activity of NPs while impacting magnetic properties. To sum up, it seems that surface functionalization could provide more potent tools for bioapplications with improving biocompatibility and bacterial inhibition of CBNPs, though; further studies are needed in this regard.

Clinical Evaluation of an HTK Solution for Liver Transplantation: A Phase 3 Randomized Pilot Clinical Trial Study.

BACKGROUND: Organ preservation solutions are not easily accessible in Iran, similar to many resource-limited countries. We aimed to evaluate the efficacy of a locally-produced HTK solution among adult liver transplantation candidates in a pilot clinical trial study. METHODS: Adult patients undergoing liver transplantation were randomly allocated into two groups. One received the HTK solution (PharMedCina Inc., Shiraz, Iran), and the second received the commercially available HTK solution (Custodiol ®). RESULTS: Overall, 28 individuals entered the study, including 11 and 9 males (78.6% and 64.3%) in the Custodiol® and local HTK groups, respectively. Clinical characteristics, including postoperative biliary complications, reperfusion syndrome, infection and primary non-function (PNF) rates, amount of intraoperative bleeding, length of hospital and ICU stay, peak aspartate aminotransferase (AST) and alanine aminotransferase (ALT), and duration of follow-up were similar between the two groups (P>0.05). One patient died in the locally-produced HTK group. The patient underwent re-transplantation 20 days after his first liver transplantation due to PNF. Two patients died in the Custodiol group, both due to PNF of the liver, which occurred five and three days after transplantation. The two groups did not show any difference regarding serum levels of AST, ALT, alkaline phosphatase (ALP), bilirubin, platelet count, prothrombin time and international normalized ratio, white blood cell count, blood urea nitrogen, and creatinine on the first postoperative day and on the day of discharge (P>0.05). CONCLUSION: Based on the findings of this pilot study with the current sample size, no statistically significant difference was found between our locally-produced HTK solution and Custodiol® regarding clinical outcomes.

Investigations of antiproliferative and antioxidant activity of ß-lactam morpholino-1,3,5-triazine hybrids.

This article reports for the first time the synthesis of some novel ß-lactam morpholino-1,3,5-triazine hybrids by a [2+2]-cycloaddition reaction of imines 7a-c, 9a-c and 11 with ketenes derived from substituted acetic acids. The reaction was totally diastereoselective, leading exclusively to the formation of cis-ß-lactams 8a-l, 10a-f and 12a-c. The synthesized compounds were tested for activity towards SW1116, MCF-7 and HepG2 cancer cell lines and non-cancerous HEK-293 cell line by MTT assay. None of the compounds exert an observable effect on HepG2, MCF-7 and HEK-293 cells, but compounds 7b, 8f, 8g, 8l, 10c, and 10e exhibited excellent growth inhibitory activity (IC50 < 5 µM) against SW 1116 cells, comparable to that of doxorubicin (IC50 = 6.9 µM). An evaluation of the antioxidant potential of each of the compounds, performed by diphenylpicrylhydrazyl (DPPH) assay, indicated that 7b, 9a, 9b and 9c have strong free radical scavenging activity. UV absorption titration studies reveal that 7b, 8l, 8g and 8f interact strongly with calf-thymus DNA (CT-DNA) in the order of 8l > 7b > 8f > 8g. Collectively, the in vitro capabilities of some of these morpholino-triazine imines and ß-lactams suggest possible applications to development of new antioxidants and DNA binding therapeutics.

Enterobacter sp. Mediated Synthesis of Biocompatible Nanostructured Iron-Polysaccharide Complexes: a Nutritional Supplement for Iron-Deficiency Anemia.

FDA has approved iron oxide nanoparticles (IONs) coated with organic compounds as a safe material with less toxic effects compared with the naked metal ions and nanoparticles. In this study, the biological and physicochemical characteristics of a nanostructured iron-polysaccharide complexes (Nano-IPC) biosynthesized by Enterobacter sp. were evaluated. Furthermore, the serum biochemical parameters, tissue iron level, red blood cell parameters, and organ ferritin of rats were measured for investigating the effect of the Nano-IPCs in comparison with FeSO4 as a supplement for iron deficiency. The biosafety data demonstrated 35% increment of viability in Hep-G2 hepatocarcinoma cell lines when treated with nanoparticles (500 µg/mL) for 24 h. Besides, iron concentration in serum and tissue as well as the expression of ferritin L subunit in animals treated with the Nano-IPCs supplement were meaningfully higher than the FeSO4-supplemented and negative control animals. Moreover, the expression level of ferritin H subunit and biochemical factors remained similar to the negative control animals in the Nano-IPC-supplemented group. These results indicated that Nano-IPCs can be considered as a nontoxic supplement for patients carrying iron-deficiency anemia (IDA).

Antimicrobial Efficacy of a Novel Antibiotic-Eluting Injectable Platelet-Rich Fibrin Scaffold against a Dual-Species Biofilm in an Infected Immature Root Canal Model.

BACKGROUND AND AIMS: This study was aimed at evaluating the antibacterial property of an injectable platelet-rich fibrin (I-PRF) scaffold containing triple antibiotic mixture against an Actinomyces naeslundii (A. naeslundii) and Enterococcus faecalis (E. faecalis) biofilm in an infected immature root canal model. METHODS: A dual-species biofilm was inoculated inside the root canals via a series of centrifugal cycles. The samples were allocated to three experimental groups (i.e., G1: triple antibiotic mixture, G2: I-PRF containing triple antibiotic mixture, and G3: antibiotic-free I-PRF scaffold) and two control groups (G4: seven-day biofilm untreated and G5: bacteria-free untreated). RESULTS: Bacterial gene quantification change and the overall reduction of live bacteria were evaluated. The highest antibacterial activity against A. naeslundii belonged to G2. However, G1 and G2 had similar antibacterial property against E. faecalis (p value = 0.814). In general, experimental groups revealed higher levels of antibacterial activity against E. faecalis than against A. naeslundii (p value < 0.001). Notably, G2 could dramatically decrease the number of live bacteria up to near 92%. CONCLUSIONS: The current study provides insight into the antibacterial property of an antibiotic-eluting I-PRF scaffold against a dual-species biofilm colonized inside the root canal. The fabricated scaffold contains not only the antibiotics but also the growth factors, which favor the regeneration.

Bacteria-assisted biogreen synthesis of radical scavenging exopolysaccharide-iron complexes: an oral nano-sized nutritional supplement with high in vivo compatibility.

Microbial exopolysaccharides (EPSs) have recently served as an efficient substrate for the production of biocompatible metal nanoparticles (NPs) given their favorable stabilizing and reducing properties due to the presence of polyanionic functional groups in their structure. In the present work, Pantoea sp. BCCS 001 GH was used to produce EPS-stabilized biogenic Fe NPs as a complex through a novel biosynthesis reaction. Physicochemical characterization of the EPS-Fe complex was performed, indicating high thermal stability, desirable magnetic properties due to the uniform distribution of the Fe NPs with the average size of ∼10 nm and spherical shape within the EPS matrix. In addition, the in vivo toxicity of the EPS-stabilized Fe NPs was evaluated to investigate their potential for the treatment of iron deficiency anemia. Biological blood parameters and organ histology studies confirmed very high safety of the biosynthesized composite, making EPS-Fe a suitable candidate with an economical and environment friendly synthesis method for a wide spectrum of potential fields in medicine.

Structural characterization of polysaccharide-coated iron oxide nanoparticles produced by Staphylococcus warneri, isolated from a thermal spring.

The biocompatible-coated iron oxide nanoparticles (IONs) have attracted a great interest because of their various applications in biological science and medicine. In most cases, the toxic effect of naked iron oxide nanoparticles is completely cleared by adding a biocompatible coating, such as polysaccharides, polyethylene glycol (PEG), or biosynthesis of biocompatible-coated IONs using microorganisms such as bacteria. In the present study, polysaccharide-coated iron oxide nanoparticles were produced by a strain of Staphylococcus warneri isolated from a thermal spring. For identification of the isolated bacterium, 16S rRNA gene sequencing was done. Characterization of the nanoparticles was performed for the first time, using transmission electron microscopy (TEM), dynamic light scattering (DLS), thermogravimetric analysis (TGA), X-ray crystallography (XRD), Fourier-transform infrared (FTIR) spectroscopy, vibrating sample magnetometer (VSM), and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Results indicated that the spherical iron oxide nanoparticles were coated by a polysaccharide (13.6%), which provided a large negative charge of -91 mV and very low saturation magnetization of around 0.28 emu/g. The result of MTT assay on MOLT-4 cell lines showed that the percentage of viability was between 95.6% and 68.9% in the 10-100 µM of nanoparticle concentrations with a high IC 50 value, which makes it appropriate for biomedical applications such as cancer therapy.

Neuroprotective Effects of Heat Shock Protein70.

BACKGROUND & OBJECTIVE: Heat Shock Proteins (HSPs) increase response to many stresses in cells. Stroke is a neural shock that leads to the destruction of a large number of brain cells, whereas induction and expression of HSPs can decrease the amount of damage, and in some conditions can cure damaged cells. HSP70 family is considered as the most important member of HSPs in normal and stress condition of cells. They are strongly up-regulated by stresses and have protective roles in under stressed cells. Therefore, in this review, we briefly consider the association between HSP70 and stroke. We searched in Pubmed and Scopus databases using the specified keywords and selected the articles based on the certain association between HSP70 and stroke. HSP70 protects cells from damage through a variety of cellular and biochemical processes such as chaperone function, anti-apoptotic, anti-necrotic and anti-inflammatory mechanisms. CONCLUSION: Protective effects of HSP70 in neurodegenerative shocks are illustrated in the review, and it can be concluded that the induction of HSP70 in stresses can be considered as a therapeutic factor, although it needs further studies.

Characterization of biogenic Fe (III)-binding exopolysaccharide nanoparticles produced by Ralstonia sp. SK03.

A new technological approach to nanoparticle synthesis is using microorganisms, such as bacteria, which have the ability to synthesize nontoxic nanoparticles with high biocompatibility. In addition, bacteria have strict control over size, structure, shape, and dimension of produced nanoparticles. In the present work, Fe (III)-binding exopolysaccharide (Fe-EPS) nanoparticles were biosynthesized by Ralstonia pickettii sp. SK03, a bacterium isolated from a mineral spring. 16S rRNA gene sequencing and biochemical tests were done for identification of the isolated bacterium. For the first time, critical biological and physicochemical properties of this iron oxide nanoparticle were characterized using Fourier Transform Infrared (FTIR) Spectroscopy, Transmission Electron Microscopy (TEM), Vibrating Sample Magnetometer (VSM), Dynamic Light Scattering (DLS), Thermogravimetric analysis (TGA), X-ray crystallography (XRD), Atomic absorption spectroscopy (AAS), and cell viability assays (MTT assay). The characterization results showed that Fe-EPS nanoparticles were composed of spherical ferrihydrite nanoparticles (with a size range of 1.2-2 nm), trapped in a polysaccharide matrix. The TGA analysis demonstrated that Fe-EPS nanoparticles contained ∼25.2% polysaccharide. Therefore, this polysaccharide matrix showed a very low magnetic saturation value (0.25 emu/g) and a large negative charge of -93.8 mV. In addition, treatment of hepatocarcinoma cell line (Hep-G2) with 1-500 µg/mL concentrations of Fe-EPS nanoparticles caused 40% increase in the cell viability, which indicated that the biosynthesized nanoparticles were nontoxic and biocompatible. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 2018 © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:1167-1176, 2018.

Harnessing self-assembled peptide nanoparticles in epitope vaccine design.

Vaccination has been one of the most successful breakthroughs in medical history. In recent years, epitope-based subunit vaccines have been introduced as a safer alternative to traditional vaccines. However, they suffer from limited immunogenicity. Nanotechnology has shown value in solving this issue. Different kinds of nanovaccines have been employed, among which virus-like nanoparticles (VLPs) and self-assembled peptide nanoparticles (SAPNs) seem very promising. Recently, SAPNs have attracted special interest due to their unique properties, including molecular specificity, biodegradability, and biocompatibility. They also resemble pathogens in terms of their size. Their multivalency allows an orderly repetitive display of antigens on their surface, which induces a stronger immune response than single immunogens. In vaccine design, SAPN self-adjuvanticity is regarded an outstanding advantage, since the use of toxic adjuvants is no longer required. SAPNs are usually composed of helical or ß-sheet secondary structures and are tailored from natural peptides or de novo structures. Flexibility in subunit selection opens the door to a wide variety of molecules with different characteristics. SAPN engineering is an emerging area, and more novel structures are expected to be generated in the future, particularly with the rapid progress in related computational tools. The aim of this review is to provide a state-of-the-art overview of self-assembled peptide nanoparticles and their use in vaccine design in recent studies. Additionally, principles for their design and the application of computational approaches to vaccine design are summarized.

Physicochemical and biological characteristics of the nanostructured polysaccharide-iron hydrogel produced by microorganism Klebsiella oxytoca.

There is an increasing interest in the nanostructured polysaccharide-iron hydrogel produced by Klebsiella oxytoca. Critical physicochemical and biological characteristics of these nanostructures should be revealed for biomedical applications. Accordingly, an iron reducing strain K. oxytoca, which synthesizes biogenic polysaccharide-iron hydrogel nanoparticles, known as Fe (III)-exopolysaccharide (Fe-EPS) was isolated from a mineral spring. For microbiological identification purpose 16S rRNA sequence analysis and different morphological, physiological, and biochemical characteristics of the isolate were studied. Critical physicochemical and biological characteristics of the produced Fe-EPS were evaluated using transmission electron microscopy (TEM), Fourier transform infrared (FTIR) spectroscopy, X-ray crystallography (XRD), vibrating sample magnetometer (VSM). In addition, for the first time, Fe-EPS which synthesized by K. oxytoca was evaluated by dynamic light scattering (DLS), thermo gravimetric analysis (TGA), and cytotoxicity assay. TEM micrographs showed that the biogenic Fe-EPS is composed of ultra-small (about 1.8 nm) iron oxide nanoparticles (IONs) which are trapped in a polysaccharide matrix. The matrix was about 17% (w/w) of Fe-EPS total weight and provided a large negative charge of -71 mV. Interestingly, Fe-EPS showed a growth promotion effect on hepatocarcinoma cell line (Hep-G2) and 36% increase in the percentage of viability was observed by 24 h exposure to 500 µg ml-1 Fe-EPS.

Optimization of Three Dimensional Culturing of the HepG2 Cell Line in Fibrin Scaffold.

BACKGROUND: A potential treatment for healing hepatic tissue is delivering isolated hepatic cells to the site of injury to promote hepatic cells formation. In this technology, providing an appropriate injectable system for delivery of hepatic cells is an important issue. In this regard, fibrin scaffolds were designed with many advantages over other scaffolds like cell delivery vehicles for biodegradation, biocompatibility and hemostasis. OBJECTIVES: The aim of this study was to determine suitable cell culture circumstances for HepG2 cell proliferation and differentiation in 3D fibrin scaffolds by evaluating Ca(2+) concentrations, cell numbers, various ratios of plasma/RPMI 1640 and thickness of fibrin scaffold. MATERIALS AND METHODS: In a one-stage experimental design, Box-Behnken design strategy was performed by Minitab 15 software (version 15, Minitab. State College, PA) with three factors at three levels (low, medium and high) and 27 runs for identification of the effects of ratio of plasma/RPMI 1640, Ca(2+) concentration and thickness on the formation of fibrin gel scaffold and 3D HepG2 culture. RESULTS: The optimal concentrations for fibrin scaffold fabrication were achieved by adding 0.15 mol CaCl2 (50 µL) and 1 × 10(5) cells to 1:4 of plasma/RPMI 1640 ratio (500 µL with 2.3 mm thickness per well). CONCLUSIONS: Our approach provided easy handle method using inexpensive materials like human plasma instead of purified fibrinogen to fabricate fibrin scaffold.

CXCL12 Modulates Prostate Cancer Cell Adhesion by Altering the Levels or Activities of ß1-Containing Integrins.

The mechanisms by which prostate cancer (PCa) cell adhesion and migration are controlled during metastasis are not well understood. Here, we studied the effect of CXCL12 in PCa cell adhesion and spreading in DU145 and PC3 cell lines using as substrates collagen I, fibronectin (FN), and their recombinant fragments. CXCL12 treatment increased ß1 integrin-dependent PC3 cell adhesion on FN which correlated with increased focal adhesion kinase activation. However neither α5ß1 nor α4ß1 subunits were involved in this adhesion. By contrast, CXCL12 decreased DU145 adhesion and spreading on FN by downregulating α5 and ß1 integrin expression. To demonstrate the clinical relevance of CXCL12 in PCa, we measured CXCL12 levels in plasma by using ELISA and found that the chemokine is elevated in PCa patients when compared to controls. The high concentration of CXCL12 in patients suffering from PCa in comparison to those with benign disease or healthy individuals implicates CXCL12 as a potential biomarker for PCa. In addition these data show that CXCL12 may be crucial in controlling PCa cell adhesion on fibronectin and collagen I, possibly via crosstalk with integrin receptors and/or altering the expression levels of integrin subunits.