Biosystems engineering of prokaryotes with tumor-killing capacities

Certain bacteria selectively attack tumor tissues and trigger tumor shrinkage by producing toxins and modulating the local immune system, but their clinical utility is limited because of the dangers posed by systemic infection. Genetic engineering can be used to minimize the risks associated with tumor-targeting pathogens, as well as to increase their efficiency in killing tumor cells. Advances in genetic circuit design have led to the development of bacterial strains with enhanced tumor-targeting capacities and the ability to secrete therapeutics, cytotoxic proteins and prodrug-cleaving enzymes, which allows their safe and effective use for cancer treatment. The present review details the recent advances in the design and application of these modified bacterial strains. © 2016 Bentham Science Publishers

Tissue distribution and correlation profiles of heavy-metal accumulation in the freshwater crayfish astacus leptodactylus

The present work details the analysis of heavy-metal and metalloid concentrations in exoskeleton, gill, hepatopancreas, and abdominal muscle tissues of 60 crayfish (Astacus leptodactylus) specimens collected from Lake HirfanlI, a dam lake located in KIrşehir (Turkey) with a low metal-contamination profile. Concentrations of 11 metals (aluminum [Al], chromium [Cd], manganese [Mn], cobalt [Co], nickel [Ni], copper [Cu], molybdenum [Mo], silver [Ag], cadmium [Cd], mercury [Hg], and lead [Pb]) and a metalloid (arsenic [As]) were measured by inductively coupled plasma-mass spectrometry, and the relative frequencies of the most abundant isotopes of Cr, Cu, Ag, Cd, Hg, and Pb were evaluated. Three correlation trends were evaluated between the following: (1) different elements in the each individual tissue, (2) individual elements in different tissues, and (3) different elements in different tissues. In addition, correlation rates of growth parameters (weight, cephalothorax length, and total length) with heavy-metal and metalloid concentrations in each tissue were investigated. Our results suggest that substantial differences in metal and metalloid-accumulation levels exist between male and female specimens, with stronger correlations between the heavy-metal concentrations observed in the male cohort. It is notable that correlation trends of Co, Cu, 52As, Cr, and Ni in exoskeleton of the male specimens display strong similarities. Likewise, a very strong correlation is present in Ni-Cd and Ni-Pb accumulations in abdominal muscle of the male specimens; a similar trend is present between Cd and Pb concentrations in the same tissue of female specimens. For correlation rates of different heavy metals and metalloid in different tissues, the strongest positive association observed was between 63Cu in gill and As in hepatopancreas, whereas the strongest negative correlation was between accumulated Ni in abdominal muscle and As in exoskeleton. Strong correlations between metals and metalloid accumulations were observed between exoskeleton and gill. In many cases, metal and metalloid accumulation was negatively correlated with growth parameters. Preferential accumulation of Cr and Cu isotopes was observed in different tissues, suggesting that significant amounts of isotope fractionation occur during heavy-metal accumulation. Relatively low correlation rates were observed between 52Cr/ 53Cr and 63Cu/65Cu concentrations in several tissue types in both male and female cohorts, whereas no such trend was observed between Cd and Pb isotopes. © 2013 Springer Science+Business Media New York

Chromium(VI) Biosorption and Bioaccumulation by Live and Acid-Modified Biomass of a Novel Morganella morganii Isolate

Conventional methods of chromium removal are often insufficient for the remediation of chromium-contaminated natural environments, necessitating the development of alternative strategies. In this paper, we report the isolation of a novel Morganella morganii strain capable of reducing hexavalent chromium to its less-toxic and less-soluble trivalent form. Cr(VI) reduction by this strain was evaluated in both acidic environments and conditions reflecting natural freshwater sources. The isolate achieved equilibrium within 3 h and displayed a specific uptake rate of 24.30 ± 1.67 mg Cr(VI)/g biomass following HCl treatment. Without acid treatment, a reduction of over 90% was recorded within 72 h for an initial Cr(VI) concentration 20 mg/L, corresponding to a Cr(VI) removal capacity of 19.36 ± 1.89 mg/g. Absorption data of acid-treated STB5 biomass most closely followed the Toth and Langmuir models. FTIR results indicate that hydroxyl groups and extracellular or cell membrane polysaccharides may be potential adsorption sites for hexavalent chromium. Our results suggest that the isolate may be used in situ for treatment of polluted freshwater environments. Copyright © Taylor & Francis Group, LLC

Nanomechanical Characterization of Osteogenic Differentiation of Mesenchymal Stem Cells on Bioactive Peptide Nanofiber Hydrogels

Stem cell differentiation is known to be influenced by the mechanical properties of the surrounding extracellular matrix (ECM); however, little is known about the mechanical phenotypes of differentiating stem cells within the ECM. Here, this study uses osteoinductive, ECM-mimetic peptide nanofibers to investigate the changes in the mechanical properties of rat mesenchymal stem cells (rMSCs) during osteogenic differentiation. In addition, octafluorocyclobutane (C4F8)-coated atomic force microscopy (AFM) cantilevers are developed to minimize tip–sample adhesion during the nanomechanical characterization of rMSCs, and osteogenic differentiation is monitored through molecular analysis in conjunction with AFM measurements. rMSCs cultured on osteoinductive peptide nanofibers differentiate at substantially higher rates, form osteogenic cell clusters, deposit calcium to the surrounding matrix, and strikingly increase their Young's moduli throughout the osteogenic differentiation process compared to controls. These results show that the elasticity profiles of differentiating rMSCs may change significantly depending on environmental factors and especially the degree of biomineralization, and that the natural elasticity responses of cells cultured on scaffolds may be considerably different from those observed on non-bioactive surfaces. This is important for the identification of cell elasticity as a biophysical marker of osteogenic differentiation of MSCs, and indicates that biomineralization might have a predominant role on cell mechanics. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinhei

Differences in the accumulation and distribution profile of heavy metals and metalloid between male and female crayfish (Astacus leptodactylus)

Concentrations of selected heavy metals and a metalloid were measured by ICP-MS in crayfish (Astacus leptodactylus) collected from Lake Hirfanli, Turkey. Aluminum (Al), chromium (52Cr, 53Cr), copper ( 63Cu, 65Cu), manganese (Mn), nickel (Ni) and arsenic (As) were measured in the exoskeleton, gills, hepatopancreas and abdominal muscle tissues of 60 crayfish of both genders. With the exception of Al, differences were determined between male and female cohorts for the accumulation trends of the above-mentioned elements in the four tissues. It was also noted that the accumulation rates of Ni and As were significantly lower in gill tissue of females compared to males and no significant difference was observed for Cu isotopes in female crayfish. Cluster Analysis (CA) recovered similar results for both genders, with links between accumulations of Ni and As being notable. Accumulation models were described separately for male and female crayfish using regression analysis, and are presented for models where R2 > 0.85. © 2013 Springer Science+Business Media New York

Removal of a reactive dye and hexavalent chromium by a reusable bacteria attached electrospun nanofibrous web

A contaminant resistant Lysinibacillus sp. NOSK was isolated from a soil sample and its Reactive Black 5 (RB5) and Cr(vi) removal efficiencies were investigated as a function of changes in the initial pH values, temperature, static/shaking conditions, reactive dye and Cr(vi) concentrations. In this study, an electrospun polysulfone nanofibrous web (PSU-NFW) was found to be effective in attachment of bacterial cells. Bacteria attached PSU-NFWs (bacteria/PSU-NFW) have shown highly efficient removal of RB5, as 99.7 ± 0.9% and 35.8 ± 0.4% for the pristine PSU-NFW. Moreover, the highest Cr(vi) removal efficiencies measured were 98.2 ± 0.6% for bacteria attached PSU-NFW and 32.6 ± 0.6% for the pristine PSU-NFW. Simultaneous removal of RB5 and Cr(vi) were also investigated. Reusability test results indicate that, bacteria/PSU-NFW can be reused for at least 7 cycles with 28.1 ± 0.6% and 66.7 ± 0.8% removal efficiencies for RB5 and Cr(vi), respectively. © Royal Society of Chemistry 2015

Screening and selection of novel animal probiotics isolated from bovine chyme

Probiotics, gut-colonizing microorganisms capable of conferring a number of health benefits to their hosts, are highly desirable as animal feed supplements. Members of the Gram-positive genus Bacillus are often utilized as probiotics, since endospores formed by those bacteria render them highly resistant to environmental extremes and therefore capable of surviving gastrointestinal tract conditions. In this study, 84 distinct bacterial colonies were obtained from bovine chyme and 29 isolates were determined as Bacillus species. These isolates were principally screened for their antimicrobial activity against a group of two Gram-positive and fourGram-negative bacteria, including known human and animal pathogens such as Salmonella enterica, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Staphylococcus aureus. Seven strains displaying strong antimicrobial activity against the test cohort were further evaluated for other properties desirable from animal probiotics, including high spore-forming capacity and adhesiveness, resistance to pH extremes and ability to form biofilms. The isolates were found to resist simulated gastrointestinal conditions and most of the antibiotics tested. In addition, plasmid presence was checked and cytotoxicity tests were performed to evaluate the potential risks of antibiotic resistance transfer and unintended pathogenic effects on host, respectively. We propose that the bacterial isolates are suitable for use as animal probiotics. © Springer-Verlag Berlin Heidelberg and the University of Milan 2012

Heparin mimetic peptide nanofiber gel promotes regeneration of full thickness burn injury

Burn injuries are one of the most common types of trauma worldwide, and their unique physiology requires the development of specialized therapeutic materials for their treatment. Here, we report the use of synthetic, functional and biodegradable peptide nanofiber gels for the improved healing of burn wounds to alleviate the progressive loss of tissue function at the post-burn wound site. These bioactive nanofiber gels form scaffolds that recapitulate the structure and function of the native extracellular matrix through signaling peptide epitopes, which can trigger angiogenesis through their affinity to basic growth factors. In this study, the angiogenesis-promoting properties of the bioactive scaffolds were utilized for the treatment of a thermal burn model. Following the excision of necrotic tissue, bioactive gels and control solutions were applied topically onto the wound area. The wound healing process was evaluated at 7, 14 and 21 days following injury through histological observations, immunostaining and marker RNA/protein analysis. Bioactive peptide nanofiber-treated burn wounds formed well-organized and collagen-rich granulation tissue layers, produced a greater density of newly formed blood vessels, and exhibited increased re-epithelialization and skin appendage development with minimal crust formation, while non-bioactive peptide nanofibers and the commercial wound dressing 3M™ Tegaderm™ did not exhibit significant efficiency over sucrose controls. Overall, the heparin-mimetic peptide nanofiber gels increased the rate of repair of burn injuries and can be used as an effective means of facilitating wound healing. © 2017 Elsevier Lt

Label-free nanometer-resolution imaging of biological architectures through surface enhanced raman scattering

Label free imaging of the chemical environment of biological specimens would readily bridge the supramolecular and the cellular scales, if a chemical fingerprint technique such as Raman scattering can be coupled with super resolution imaging. We demonstrate the possibility of label-free super-resolution Raman imaging, by applying stochastic reconstruction to temporal fluctuations of the surface enhanced Raman scattering (SERS) signal which originate from biomolecular layers on large-area plasmonic surfaces with a high and uniform hot-spot density (>1011/cm2, 20 to 35 nm spacing). A resolution of 20 nm is demonstrated in reconstructed images of self-assembled peptide network and fibrilated lamellipodia of cardiomyocytes. Blink rate density is observed to be proportional to the excitation intensity and at high excitation densities (>10 kW/cm2) blinking is accompanied by molecular breakdown. However, at low powers, simultaneous Raman measurements show that SERS can provide sufficient blink rates required for image reconstruction without completely damaging the chemical structure

Interactions between metals accumulated in the narrow-clawed crayfish Astacus leptodactylus (Eschscholtz, 1823) in Dikilitaş Lake, Turkey

The accumulations of Al, Cd, Cr, Cu, Fe, Ni, Pb and Zn in the exoskeleton, gills, hepatopancreas and abdominal muscles of crayfish Astacus leptodactylus (Eschscholtz, 1823) were determined. The strongest correlation observed was between Cr and Ni in the gills (r = 0.904); moderate to strong correlations between Al, Cr, Fe, Ni and Cu were also observed in gill tissue. Disregarding the gills, the strongest correlation was found between Cu and Zn in the hepatopancreas (r = 0.808); the correlation between these two metals might have been a result of metallothionein activity. The accumulation of Pb was found to correlate with that of Cd in the exoskeleton, Cd and Zn in the gills, Zn and Cu in the hepatopancreas and Cu in the abdominal muscle. None of these correlations were present in lakewater and sediment samples, suggesting that the crayfish metabolism may be responsible for the co-accumulation of metal–metal pairs. As all correlations in non-gill tissues are observed between divalent metals, a shared transporter such as divalent metal transporter 1 might be involved in the accumulation of these metals. © 2015 Taylor & Francis