Six environmental fosmid clones from Antarctic coastal water bacterioplankton were completely sequenced. The genome fragments harbored small-subunit rRNA genes that were between 85 and 91% similar to those of their nearest cultivated relatives. The six fragments span four phyla, including the Gemmatimonadetes, Proteobacteria (␣ and ␥), Bacteroidetes, and high-G؉C gram-positive bacteria. Gene-finding and annotation analyses identified 244 total open reading frames. Amino acid comparisons of 123 and 113 Antarctic bacterial amino acid sequences to mesophilic homologs from G؉C-specific and SwissProt/UniProt databases, respectively, revealed widespread adaptation to the cold. The most significant changes in these Antarctic bacterial protein sequences included a reduction in salt-bridge-forming residues such as arginine, glutamic acid, and aspartic acid, reduced proline contents, and a reduction in stabilizing hydrophobic clusters. Stretches of disordered amino acids were significantly longer in the Antarctic sequences than in the mesophilic sequences. These characteristics were not specific to any one phylum, COG role category, or G؉C content and imply that underlying genotypic and biochemical adaptations to the cold are inherent to life in the permanently subzero Antarctic waters.

Hydrothermal vent ecosystems support diverse life forms, many of which rely on symbiotic associations to perform functions integral to survival in these extreme physicochemical environments. Epsilonproteobacteria, found free-living and in intimate associations with vent invertebrates, are the predominant vent-associated microorganisms. The vent-associated polychaete worm, Alvinella pompejana, is host to a visibly dense fleece of episymbionts on its dorsal surface. The episymbionts are a multispecies consortium of Epsilonproteobacteria present as a biofilm. We unraveled details of these enigmatic, uncultivated episymbionts using environmental genome sequencing. They harbor wide-ranging adaptive traits that include high levels of strain variability analogous to Epsilonproteobacteria pathogens such as Helicobacter pylori, metabolic diversity of free-living bacteria, and numerous orthologs of proteins that we hypothesize are each optimally adapted to specific temperature ranges within the 10 -65°C fluctuations characteristic of the A. pompejana habitat. This strategic combination enables the consortium to thrive under diverse thermal and chemical regimes. The episymbionts are metabolically tuned for growth in hydrothermal vent ecosystems with genes encoding the complete rTCA cycle, sulfur oxidation, and denitrification; in addition, the episymbiont metagenome also encodes capacity for heterotrophic and aerobic metabolisms. Analysis of the environmental genome suggests that A. pompejana may benefit from the episymbionts serving as a stable source of food and vitamins. The success of Epsilonproteobacteria as episymbionts in hydrothermal vent ecosystems is a product of adaptive capabilities, broad metabolic capacity, strain variance, and virulent traits in common with pathogens.Epsilonproteobacteria ͉ hydrothermal vent T he polychaete Alvinella pompejana is an endemic inhabitant of deep-sea hydrothermal vents located from 21°N to 32°S latitude on the East Pacific Rise (1). This tube-dwelling polychaete forms dense colonies exclusively on the walls of high-temperature black smoker chimneys (2, 3), which are characterized by extreme physicochemical gradients and dynamic in thermal emission rates and intensive mineral precipitation. The high-temperature diffuse flow surrounding the worms' tubes is acidic (pH 4.2-6.1) carrying high levels of total (free ϩ complexed) hydrogen sulfide (Ͼ1 mM), ammonia (3.8-10 m), and reactive heavy metals (0.3-200 M) including ferrous iron (290-840 m) (2). Temperature fluctuations in the actual tubes of A. pompejana range from 29°C to 84°C while the chemical conditions are anoxic, slightly acidic to near neutral pH (5.33-6.9), and rich in electron acceptors (sulfate, nitrate, Fe III, and Mg) as well as potentially lethal levels of heavy metals (2, 3). The tube fluids contain surprisingly low levels of free H 2 S (Ͻ0.2 M to 46.53 M) and are a mix of ambient seawater (72-

Herein, the NH2-UiO-66 metal organic framework (MOF) has been green synthesized with the assistance of high gravity to provide a suitable and safe platform for drug loading. The NH2-UiO-66 MOF was characterized using a field-emission scanning electron microscope, transmission electron microscope (TEM), X-ray diffraction, and zeta potential analysis. Doxorubicin was then encapsulated physically on the porosity of the green MOF. Two different stimulus polymers, p­(HEMA) and p­(NIPAM), were used as the coating agents of the MOFs. Doxorubicin was loaded onto the polymer-coated MOFs as well, and a drug payload of more than 51% was obtained, which is a record by itself. In the next step, pCRISPR was successfully tagged on the surface of the modified MOFs, and the performance of the final nanosystems were evaluated by the GFP expression. In addition, successful loadings and internalizations of doxorubicin were investigated via confocal laser scanning microscopy. Cellular images from the HeLa cell line for the UiO-66@DOX@pCRISPR and GMA-UiO-66@DOX@pCRISPR do not show any promising and successful gene transfections, with a maximum EGFP of 1.6%; however, the results for the p­(HEMA)-GMA-UiO-66@DOX@pCRISPR show up to 4.3% transfection efficiency. Also, the results for the p­(NIPAM)-GMA-UiO-66@DOX@pCRISPR showed up to 6.4% transfection efficiency, which is the first and superior report of a MOF-based nanocarrier for the delivery of pCRISPR. Furthermore, the MTT assay does not shown any critical cytotoxicity, which is a promising result for further biomedical applications. At the end of the study, the morphologies of all of the nanomaterials were screened after drug and gene delivery procedures and showed partial degradation of the nanomaterial. However, the cubic structure of the MOFs has been shown in TEM, and this is further proof of the stability of these green MOFs for biomedical applications.

Cardiovascular diseases are the number one cause of heart failure and death in the world, and the transplantation of the heart is an effective and viable choice for treatment despite presenting many disadvantages (most notably, transplant heart availability). To overcome this problem, cardiac tissue engineering is considered a promising approach by using implantable artificial blood vessels, injectable gels, and cardiac patches (to name a few) made from biodegradable polymers. Biodegradable polymers are classified into two main categories: natural and synthetic polymers. Natural biodegradable polymers have some distinct advantages such as biodegradability, abundant availability, and renewability but have some significant drawbacks such as rapid degradation, insufficient electrical conductivity, immunological reaction, and poor mechanical properties for cardiac tissue engineering. Synthetic biodegradable polymers have some advantages such as strong mechanical properties, controlled structure, great processing flexibility, and usually no immunological concerns; however, they have some drawbacks such as a lack of cell attachment and possible low biocompatibility. Some applications have combined the best of both and exciting new natural/ synthetic composites have been utilized. Recently, the use of nanostructured polymers and polymer nanocomposites has revolutionized the field of cardiac tissue engineering due to their enhanced mechanical, electrical, and surface properties promoting tissue growth. In this review, recent research on the use of biodegradable natural/synthetic nanocomposite polymers in cardiac tissue engineering is presented with forward looking thoughts provided for what is needed for the field to mature.

Introduction: In recent years, the use of cost-effective, multifunctional, environmentally friendly and simple prepared nanomaterials/nanoparticles have been emerged considerably. In this manner, different synthesizing methods were reported and optimized, but there is still lack of a comprehensive method with multifunctional properties. Materials and Methods: In this study, we aim to synthesis the copper oxide nanoparticles using Achillea millefolium leaf extracts for the first time. Catalytic activity was investigated by in situ azide alkyne cycloaddition click and also A 3 coupling reaction, and optimized in terms of temperature, solvent, and time of the reaction. Furthermore, the photocatalytic activity of the synthesized nanoparticles was screened in terms of degradation methylene blue dye. Biological activity of the synthesized nanoparticles was evaluated in terms of antibacterial and anti-fungal assessments against Staphylococcus aureus, M. tuberculosis, E. coli, K. pneumoniae, P. mirabili, C. diphtheriae and S. pyogenes bacteria's and G. albicans, A. flavus, M. canis and G. glabrata fungus. In the next step, the biosynthesized CuO-NPs were screened by MTT and NTU assays. Results: Based on our knowledge, this is a comprehensive study on the catalytic and biological activity of copper oxide nanoparticles synthesizing from Achillea millefolium, which presents great and significant results (in both catalytic and biological activities) based on a simple and green procedure. Conclusion: Comprehensive biomedical and catalytic investigation of the biosynthesized CuO-NPs showed the mentioned method leads to synthesis of more eco-friendly nanoparticles. The in vitro studies showed promising and considerable results, and due to the great stability of these nanoparticles in a green media, effective biological activity considered as an advantageous.

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Hybrid bioactive inorganic−organic carbon-based nanocomposites of reduced graphene oxide (rGO) nanosheets enlarged with multi-walled carbon nanotubes (MWCNTs) were decorated to provide a suitable space for in situ growth of CoNi 2 S 4 and green-synthesized ZnO nanoparticles. The ensuing nanocarrier supplied π−π interactions between the DOX drug and a stabilizing agent derived from leaf extracts on the surface of ZnO nanoparticles and hydrogen bonds; gene delivery of (p)CRISPR was also facilitated by chitosan and alginate renewable macromolecules. Also, these polymers can inhibit the potential interactions between the inorganic parts and cellular membranes to reduce the potential cytotoxicity. Nanocomposite/nanocarrier analyses and sustained DOX delivery (cytotoxicity analyses on HEK-293, PC12, HepG2, and HeLa cell lines after 24, 48, and 72 h) were indicative of an acceptable cell viability of up to 91.4 and 78.8% after 48 at low and high concentrations of 0.1 and 10 μg/mL, respectively. The MTT results indicate that by addition of DOX to the nanostructures, the relative cell viability increased after 72 h of treatment; since the inorganic compartments, specifically CoNi 2 S 4 , are toxic, this is a promising route to increase the bioavailability of the nanocarrier before reaching the targeted cells. Nanosystems were tagged with (p)CRISPR for co-transfer of the drug/genes, where confocal laser scanning microscopy (CLSM) pictures of the 4′,6-diamidino-2phenylindole (DAPI) were indicative of appropriate localization of DOX into the nanostructure with effective cell and drug delivery at varied pH. Also, the intrinsic toxicity of CoNi 2 S 4 does not affect the morphology of the cells, which is a breakthrough. Furthermore, the CLSM images of the HEK-293 and HeLa cell displayed effective transport of (p)CRISPR into the cells with an enhanced green fluorescent protein (EGFP) of up to 8.3% for the HEK-293 cell line and 21.4% for the HeLa cell line, a record. Additionally, the specific morphology of the nanosystems before and after the drug/gene transport events, via images by TEM and FESEM, revealed an intact morphology for these biopolymers and their complete degradation after long-time usage.

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