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Interest in the extraction of phytochemical bioactive compounds, especially polyphenols from biomass, has recently increased due to their valuable biological potential as natural sources of antioxidants, which could be used in a wide range of applications, from foods and pharmaceuticals to green polymers and bio-based materials. The present research study aimed to provide a comprehensive chemical characterization of the phytochemical composition of forest biomass (bark and needles) of softwood species (Picea abies L., H. Karst., and Abies alba Mill.) and to investigate their in vitro antioxidant and antimicrobial activities to assess their potential in treating and healing infected chronic wounds. The DPPH radical-scavenging method and P-LD were used for a mechanistic explanation of the biomolecular effects of the investigated bioactive compounds. (+)-Catechin, epicatechin, rutin, myricetin, 4 hydroxybenzoic and p-cumaric acids, kaempherol, and apigenin were the main quantified polyphenols in coniferous biomass (in quantities around 100 mu g/g). Also, numerous phenolic acids, flavonoids, stilbenes, terpenes, lignans, secoiridoids, and indanes with antioxidant, antimicrobial, anti-inflammatory, antihemolytic, and anti-carcinogenic potential were identified. The Abies alba needle extract was more toxic to microbial strains than the eukaryotic cells that provide its active wound healing principles. In this context, developing industrial upscaling strategies is imperative for the long-term success of biorefineries and incorporating them as part of a circular bio-economy.

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Multidrug-resistant Gram-negative bacteria such as Acinetobacter baumannii, Pseudomonas aeruginosa, and members of the Enterobacterales order are a challenging multi-sectorial and global threat, being listed by the WHO in the priority list of pathogens requiring the urgent discovery and development of therapeutic strategies. We present here an overview of the antibiotic resistance profiles and epidemiology of Gram-negative pathogens listed in the ESCAPE group circulating in Romania. The review starts with a discussion of the mechanisms and clinical significance of Gram-negative bacteria, the most frequent genetic determinants of resistance, and then summarizes and discusses the epidemiological studies reported for A. baumannii, P. aeruginosa, and Enterobacterales-resistant strains circulating in Romania, both in hospital and veterinary settings and mirrored in the aquatic environment. The Romanian landscape of Gram-negative pathogens included in the ESCAPE list reveals that all significant, clinically relevant, globally spread antibiotic resistance genes and carrying platforms are well established in different geographical areas of Romania and have already been disseminated beyond clinical settings.

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The current paper estimated the physico-chemical properties of water in the Danube Delta (Romania), based on Sentinel 2 remote sensing data. Eleven sites from the Danube Delta were sampled in spring and autumn for three years (2018-2020) and 21 water physico-chemical parameters were measured in laboratory. Several families of machine learning algorithms, translated into hundreds of models with different parameterizations for each machine learning algorithm, based on remote sensing data input from Sentinel 2 spectral bands, were employed to find the best models that predicted the values measured in laboratory. This was a novel approach, reflected in the types of selected models that minimised the values of performance metrics for the tested parameters. For alkalinity, calcium, chloride, carbon dioxide, hardness, potassium, sodium, ammonium, dissolved oxygen, sulphates, and suspended matter the results were promising, with an overall percentage bias of the estimates of +/- 10% from the observed values. For copper, magnesium, nitrites, nitrates, turbidity and zinc the estimates were fairly accurate, with percentage biases in the interval +/- 10% - 20%, whereas for detergents, led, and phosphates the percentage bias was higher than 20%. Overall, the results of the current study showed fairly good estimates between remote sensing based estimates and laboratory measured values for most water physico-chemical parameters.

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The aim of this study was to establish the best ultrasound assisted extraction (UAE) conditions of saponins from Hedera helix L. leaves and to evaluate the in vitro biocompatibility of the extracts richest in saponins. Different parameters, such as extraction time, temperature, ultrasound power, solvent to plant material ratio, and solvent concentration, were investigated. The most efficient extraction conditions were a temperature of 50 degrees C, an ultrasound amplitude of 40%, an extraction time of 60 min, a plant material to solvent ratio of 1:20 (w:v), and 80% ethanol as solvent. In vitro cytotoxicity of the extracts richest in saponins and their influence on the DNA content of L929 (NCTC) fibroblasts were tested. Until 200 mu g/mL, the studied extracts were cytocompatible with L929 fibroblast cell lines at 48 h of treatment. These in vitro cell culture results provide useful information for further applications of Hedera helix extracts in a pharmaceutical field.

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To better understand the potential of drug repurposing in COVID-19, we analyzed control strategies over essential host factors for SARS-CoV-2 infection. We constructed comprehensive directed protein-protein interaction (PPI) networks integrating the top-ranked host factors, the drug target proteins and directed PPI data. We analyzed the networks to identify drug targets and combinations thereof that offer efficient control over the host factors. We validated our findings against clinical studies data and bioinformatics studies. Our method offers a new insight into the molecular details of the disease and into potentially new therapy targets for it. Our approach for drug repurposing is significant beyond COVID-19 and may be applied also to other diseases.

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Background: Romania is one of the European countries reporting very high antimicrobial resistance (AMR) rates and consumption of antimicrobials. We aimed to characterize the AMR profiles and clonality of 304 multi-drug resistant (MDR) Acinetobacter baumannii (Ab) and Pseudomonas aeruginosa (Pa) strains isolated during two consecutive years (2018 and 2019) from hospital settings, hospital collecting sewage tanks and the receiving wastewater treatment plants (WWTPs) located in the main geographical regions of Romania. Methods: The strains were isolated on chromogenic media and identified by MALDI-TOF-MS. Antibiotic susceptibility testing and confirmation of ESBL- and CP- producing phenotypes and genotypes were performed. The genetic characterization also included horizontal gene transfer experiments, whole-genome sequencing (WGS), assembling, annotation and characterization. Results: Both clinical and aquatic isolates exhibited high MDR rates, especially the Ab strains isolated from nosocomial infections and hospital effluents. The phenotypic resistance profiles and MDR rates have largely varied by sampling point and geographic location. The highest MDR rates in the aquatic isolates were recorded in Galati WWTP, followed by Bucharest. The Ab strains harbored mostly bla(OX)(A-)(23), bla(OXA-)(24), bla(SHV), bla(TEM) and bla(GES), while Pa strains bla(IMP), bla(VIM), bla(NDM), bla(V)(EB), bla(GES) and bla(TEM), with high variations depending on the geographical zone and the sampling point. The WGS analysis revealed the presence of antibiotic resistance genes (ARGs) to other antibiotic classes, such as aminoglycosides, tetracyclines, sulphonamides, fosfomycin, phenicols, trimethoprim-sulfamethoxazole as well as class 1 integrons. The molecular analyses highlighted: (i) The presence of epidemic clones such as ST2 for Ab and ST233 and ST357 for Pa; (ii) The relatedness between clinical and hospital wastewater strains and (iii) The possible dissemination of clinical Ab belonging to ST2 (also proved in the conjugation assays for bla(OX)(A-)(23) or bla(OX)(A-)(72) genes), ST79 and ST492 and of Pa strains belonging to ST357, ST640 and ST621 in the wastewaters. Conclusion: Our study reveals the presence of CP-producing Ab and Pa in all sampling points and the clonal dissemination of clinical Ab ST2 strains in the wastewaters. The prevalent clones were correlated with the presence of class 1 integrons, suggesting that these isolates could be a significant reservoir of ARGs, being able to persist in the environment.

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Impact on aquatic ecosystems from intensive agriculture can be mitigated through a variety of structural and management conservation practices. This article provides an overview of the main Best Management Practices (BMP) for reducing direct or indirect detrimental impact from agriculture on inland waters’ quality and ecological status. The BMP's have been shown to be more efficient compared to conventional ones and comprise a set of recommendations that should be considered carefully by environmental scientists, stakeholders, farmers and land-owners alike. © 2022 Elsevier Inc. All rights reserved

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Current advances in computational modelling and simulation have led to the inclusion of computer scientists as partners in the process of engineering of new nanomaterials and nanodevices. This trend is now, more than ever, visible in the field of deoxyribonucleic acid (DNA)-based nanotechnology, as DNA's intrinsic principle of self-assembly has been proven to be highly algorithmic and programmable. As a raw material, DNA is a rather unremarkable fabric. However, as a way to achieve patterns, dynamic behavior, or nano-shape reconstruction, DNA has been proven to be one of the most functional nanomaterials. It would thus be of great potential to pair up DNA's highly functional assembly characteristics with the mechanic properties of other well-known bio-nanomaterials, such as graphene, cellulos, or fibroin. In the current study, we perform projections regarding the structural properties of a fibril mesh (or filter) for which assembly would be guided by the controlled aggregation of DNA scaffold subunits. The formation of such a 2D fibril mesh structure is ensured by the mechanistic assembly properties borrowed from the DNA assembly apparatus. For generating inexpensive pre-experimental assessments regarding the efficiency of various assembly strategies, we introduced in this study a computational model for the simulation of fibril mesh assembly dynamical systems. Our approach was based on providing solutions towards two main circumstances. First, we created a functional computational model that is restrictive enough to be able to numerically simulate the controlled aggregation of up to 1000s of elementary fibril elements yet rich enough to provide actionable insides on the structural characteristics for the generated assembly. Second, we used the provided numerical model in order to generate projections regarding effective ways of manipulating one of the the key structural properties of such generated filters, namely the average size of the openings (gaps) within these meshes, also known as the filter's aperture. This work is a continuation of Amarioarei et al., 2018, where a preliminary version of this research was discussed.

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Subterranean streams represent unique heterotrophic ecosystems, usually supported by organic matter imported from the surface. Traditionally, the biological communities from subterranean streams were characterized as simple associations, with low diversity and species abundance, comprising mostly aquatic invertebrates connected by few trophic links compared with those of the surface. However, these features have not yet been described in the wider context of fluxes of energy and nutrients through food webs along a gradual switch from autotrophy (dominated by photosynthesis) towards heterotrophy (dominated by detritus) following the surface-subterranean continuum. Combining the most recent predictions of Ecological Stoichiometry and the Metabolic Theory of Ecology, this article provides a theoretical framework aiming to explain the patterns observed along the surface-subterranean continuum in streams. It is predicted that the main factors constraining the structure and functioning of communities and food webs are the decline in the quantity and diversity of basal resources along this gradient, along with nutrients availability in water that affects food quality. With increasing availability of dissolved nutrients in water, sinking-cave streams are hypothesized to fluctuate between being N and/ or P co-limited to C-limited. Combined, the quantity, quality, and diversity of basal resources regulate subterranean aquatic communities through bottom-up mechanisms, reflected in a decreased flux of macronutrients through food webs. The consequences of these bottom-up effects are decreased abundance, biomass, secondary production, consumption rate, and mean body size of communities, together with potential increases in the elemental imbalance for macronutrients, omnivory, trophic position, and niche width and overlap among aquatic consumers along the surface-subterranean continuum. The bottom-up effects induce changes in the topology of stream food webs, which become shorter, with lower trophic diversity at the base of the network, but increased connectance along this environmental gradient.

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Chronic infections represent an important burden on the healthcare system and have a significant impact on the patients' quality of life. While Staphylococcus spp. are commensal bacteria, they can become pathogenic, leading to various types of infections. In this study we aimed to characterize the virulence profiles of staphylococcal strains involved in difficult-to-treat skin and soft tissue infections, from both phenotypic and genotypic points of view. Phenotypic ability of the strains to secrete soluble virulence factors was assessed by a culturing dependent assay and their capacity to develop biofilms on inert substrate was screened by an adapted crystal violet microtiter method. We also tested the presence of several virulence genes by PCR. Most of the studied strains were isolated from purulent secretions of acne lesions and frequently secreted two or three soluble virulence factors. Most frequently secreted soluble virulence factors were caseinase (89%), lipase (71%) and lecithinase (67%). Almost half of the strains produced a well-represented biofilm. The molecular characterization showed the presence of the genes cna, hlg, clfA, and clfB. Staphylococcal strains that produce difficult-to-treat skin and soft tissue infections seem to be characterized by an enhanced ability to produce different soluble virulence factors and to develop biofilms in vitro. Further studies need to be developed in other Staphylococcus spp. infections in order to confirm this hypothesis.

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In this paper we describe the transmission of a multi-drug resistant Klebsiella pneumoniae ST101 clone from hospital to wastewater and its persistence after chlorine treatment. Water samples from influents and effluents of the sewage tank of an infectious diseases hospital and clinical strains collected from the intra-hospital infections, during a period of 10 days prior to wastewater sampling were analyzed. Antibiotic resistant K. pneumoniae strains from wastewaters were recovered on selective media. Based on antibiotic susceptibility profiles and PCR analyses of antibiotic resistance (AR) genetic background, as well as whole-genome sequencing (Illumina MiSeq) and subsequent bioinformatic analyses, 11 ST101 K. pneumoniae strains isolated from hospital wastewater influent, wastewater effluent and clinical sector were identified as clonally related. The SNP and core genome analyses pointed out that five strains were found to be closely related (with <= 18 SNPs and identical cgMLST profile). The strains belonging to this clone harbored multiple acquired AR genes [bla(CTX-M-)(15), bla(OXA-)(48), bla(OXA-)(1), bla(SHV-)(106), bla(TEM-)(150), aac(3)-IIa, aac(6 ')-Ib-cr, oqxA10, oqxB17, fosA, catB3, dfrA14, tet(D)] and chromosomal mutations involved in AR (Delta mgrB, Delta ompK35, amino acid substitutions in GyrA Ser83Tyr, Asp87Asn, ParC Ser80Tyr). Twenty-nine virulence genes involved in iron acquisition, biofilm and pili formation, adherence, and the type six secretion system - T6SS-III were identified. Our study proves the transmission of MDR K. pneumoniae from hospital to the hospital effluent and its persistence after the chlorine treatment, raising the risk of surface water contamination and further dissemination to different components of the trophic chain, including humans.

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Acinetobacter baumannii has emerged worldwide as a dominant pathogen in a broad range of severe infections, raising an acute need for efficient antibacterials. This is the first report on the resistome and virulome of 33 extended drug-resistant and carbapenem-resistant A. baumannii (XDR CRAB) strains isolated from hospitalized and ambulatory patients in Bucharest, Romania. A total of 33 isolates were collected and analyzed using phenotypic antibiotic susceptibility and conjugation assays, PCR, whole-genome sequencing (WGS), pulsed-field gel electrophoresis (PFGE) and MultiLocus Sequence Typing (MLST). All isolates were extensively drug-resistant (XDR), being susceptible only to colistin. The carbapenem resistance was attributed by PCR mainly to blaOXA-24 and blaOXA-23 genes. PFGE followed by MLST analysis demonstrated the presence of nine pulsotypes and six sequence types. WGS of seven XDR CRAB isolates from healthcare-associated infections demonstrated the high diversity of resistance genes repertoire, as well as of mobile genetic elements, carrying ARGs for aminoglycosides, sulphonamides and macrolides. Our data will facilitate the understanding of resistance, virulence and transmission features of XDR AB isolates from Romanian patients and might be able to contribute to the implementation of appropriate infection control measures and to develop new molecules with innovative mechanisms of action, able to fight effectively against these bugs, for limiting the spread and decreasing the infection rate and mortality. © 2021, The Author(s).

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Subterranean streams represent unique heterotrophic ecosystems, usually supported by organic matter imported from the surface. Traditionally, the biological communities from subterranean streams were characterized as simple associations, with low diversity and species abundance, comprising mostly aquatic invertebrates connected by few trophic links compared with those of the surface. However, these features have not yet been described in the wider context of fluxes of energy and nutrients through food webs along a gradual switch from autotrophy (dominated by photosynthesis) towards heterotrophy (dominated by detritus) following the surface–subterranean continuum. Combining the most recent predictions of Ecological Stoichiometry and the Metabolic Theory of Ecology, this article provides a theoretical framework aiming to explain the patterns observed along the surface–subterranean continuum in streams. It is predicted that the main factors constraining the structure and functioning of communities and food webs are the decline in the quantity and diversity of basal resources along this gradient, along with nutrients availability in water that affects food quality. With increasing availability of dissolved nutrients in water, sinking-cave streams are hypothesized to fluctuate between being N and/ or P co-limited to C-limited. Combined, the quantity, quality, and diversity of basal resources regulate subterranean aquatic communities through bottom–up mechanisms, reflected in a decreased flux of macronutrients through food webs. The consequences of these bottom–up effects are decreased abundance, biomass, secondary production, consumption rate, and mean body size of communities, together with potential increases in the elemental imbalance for macronutrients, omnivory, trophic position, and niche width and overlap among aquatic consumers along the surface–subterranean continuum. The bottom–up effects induce changes in the topology of stream food webs, which become shorter, with lower trophic diversity at the base of the network, but increased connectance along this environmental gradient. © 2021, The Author(s), under exclusive licence to Springer Nature Switzerland AG.

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Antimicrobial resistance (AMR) is a significant global threat to both public health and the environment. The emergence and expansion of AMR is sustained by the enormous diversity and mobility of antimicrobial resistance genes (ARGs). Different mechanisms of horizontal gene transfer (HGT), including conjugation, transduction, and transformation, have facilitated the accumulation and dissemination of ARGs in Gram-negative and Gram-positive bacteria. This has resulted in the development of multidrug resistance in some bacteria. The most clinically significant ARGs are usually located on different mobile genetic elements (MGEs) that can move intracellularly (between the bacterial chromosome and plasmids) or intercellularly (within the same species or between different species or genera). Resistance plasmids play a central role both in HGT and as support elements for other MGEs, in which ARGs are assembled by transposition and recombination mechanisms. Considering the crucial role of MGEs in the acquisition and transmission of ARGs, a potential strategy to control AMR is to eliminate MGEs. This review discusses current progress on the development of chemical and biological approaches for the elimination of ARG carriers.

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Carrier red blood cells (RBCs) can be used for vascular delivery o f encapsulated drugs, biologicals and nanoparticies o f various compositions with multiple advantages including bioavailability, biocompatibility, large carrier volumes and vascular residence time. Iron-based magnetic nanoparticies (MNPs), with size less than 15 nm became superparamagnetic and they can be used for both, diagnostic (imagistic) and therapeutic purposes, including as controlled release drug delivery systems. For these reasons, the objective o f this study was to employ human erythrocytes for the encapsulation of carbon shell iron-based core nanoparticies (obtained by continuous wave infrared laser pyrolysis o f Fe(CO)(5) by a transient opening o f cell membrane pores using different hypo-osmotic dialysis. In order to evaluate the encapsulation o f nanoparticies into red blood cells (RBCs), normal, unloaded and loaded red blood cells (RBCs) were examined by Scanning/Transmission Electron Microscopy and Energy-dispersive X-ray spectroscopy analysis (EDX). Here, it is reported that these composite iron-based nanoparticies that were synthesized and tested could lead to their use in conjunction with the red blood cells (RBCs) as a promising agent in biomedical applications.

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Since its introduction in the early 2000s, DNA origami had a big impact on the development of nanotechnology by gathering numerous applications. During this time, many tools were designed and used to generate arbitrary shapes capable of self-assembly which make this technique more approachable. In this paper, we have created the map of Romania at nanoscale dimensions by using a new open-source software - PERDIX. For this purpose, we used a scaffold strand with a length of 6959 nucleotides and 162 staple strands with a variable length ranging between 20 and 63 nucleotides. All the computational tools that were used in this experiment are open-source and user-friendly.

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Anemia is one of the most common disorders in the world generated by multiple causes. Despite a good understanding of the causes of anemia, there is still uncertainty about how best this should be investigated, prevented and managed. This reflects the limitations of laboratory tests, as well as the poor understanding of the complex physiological mechanisms. A better understanding of the nature of anemia and the changes in erythrocytes may provide new strategies to benefits of patients. In this study, it was identified by flow cytometric and SEM analysis, the morphological changes of forms that deviate from the classical discoid shape, which had an increase in the mean fluorescence intensity of calcein (erythrocyte viability measured by the level of intracellular esterase activity with the Calcein-AM method). The results obtained for RBCs from the patients with different types of anemia did not demonstrate a massive phosphatidylserine externalization measurable with Annexin-V-FITC compared to the values for normal erythrocytes. The present study provides that flow cytometric analysis can be a quick and efficient method for diagnosing in haematological diseases and monitoring and in the medical biotechnology industry in order to develop drugs to treat anemia.

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We report on the genomic characterization of 47 multi-drug resistant, carbapenem resistant and ESBL-producing K. pneumoniae isolates from the influent (I) and effluent (E) of three wastewater treatment plants (WWTPs) and from Romanian hospital units which are discharging the wastewater in the sampled WWTPs. The K. pneumoniae whole genome sequences were analyzed for antibiotic resistance genes (ARGs), virulence genes and sequence types (STs) in order to compare their distribution in C, I and E samples. Both clinical and environmental samples harbored prevalent and widely distributed ESBL genes, i.e. blaSHV, bla(OXA), bla(TEM) and bla(CTX M). The most prevalent carbapenemase genes were bla(NDM-1), bla(OXA-48) and bla(KPC-2). They were found in all types of isolates, while bla(OXA-162), a rare blaOXA-48 variant, was found exclusively in water samples. A higher diversity of carbapenemases genes was seen in wastewater isolates. The aminoglycoside modifying enzymes (AME) genes found in all types of samples were aac(6'), ant(2'')Ia, aph(3'), aaD, aac(3) and aph(6). Quinolone resistance gene qnrS1 and the multi-drug resistance oqxA/B pump gene were found in all samples, while qnrD and qnrB were associated to aquatic isolates. The antiseptics resistance gene qacEdelta1 was found in all samples, while qacE was detected exclusively in the clinical ones. Trimethroprim-sulfamethoxazole (dfrA, sul1 and sul2), tetracyclines (tetA and tetD) and fosfomycin (fosA6, known to be located on a transpozon) resistance genes were found in all samples, while for choramphenicol and macrolides some ARGs were detected in all samples (catA1 and catB3 / mphA), while other (catA2, cmIA5 and aac(6')Ib / mphE and msrE) only in wastewater samples. The rifampin resistance genes arr2 and 3 (both carried by class I integrons) were detected only in water samples. The highly prevalent ARGs preferentially associating with aquatic versus clinical samples could ascribe potential markers for the aquatic (blaSHV-145, qacEdelta1, sul1, aadA1, aadA2) and clinical (bla(OXA-1), bla(SHV-106),bla(TEM-150), aac(3)Iia, dfrA14, oqxA10; oqxB17,catB3, tetD) reservoirs of AR. Moreover, some ARGs (oqxA10; blaSHV-145; blaSHV-100, aac(6')Il, aph(3') VI, armA, arr2, cmlA5, blaCMY-4, mphE, msrE, oqxB13, blaOXA-10) showing decreased prevalence in influent versus effluent wastewater samples could be used as markers for the efficiency of the WWTPs in eliminating AR bacteria and ARGs. The highest number of virulence genes (75) was recorded for the I samples, while for E and C samples it was reduced to half. The most prevalent belong to three functional groups: adherence (fim genes), iron acquisition (ent, fep, fyu, irp and ybt genes) and the secretion system (omp genes). However, none of the genes associated with hypervirulent K. pneumoniae have been found. A total of 14 STs were identified. The most prevalent clones were ST101, ST219 in clinical samples and ST258, ST395 in aquatic isolates. These STs were also the most frequently associated with integrons. ST45 and ST485 were exclusively associated with I samples, ST11, ST35, ST364 with E and ST1564 with C samples. The less frequent ST17 and ST307 aquatic isolates harbored blaOXA-162, which was co-expressed in our strains with bla(CTX-M-15) and bla(OXA-1).

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We report on the genomic characterization of 47 multi-drug resistant, carbapenem resistant and ESBL-producing K. pneumoniae isolates from the influent (I) and effluent (E) of three wastewater treatment plants (WWTPs) and from Romanian hospital units which are discharging the wastewater in the sampled WWTPs. The K. pneumoniae whole genome sequences were analyzed for antibiotic resistance genes (ARGs), virulence genes and sequence types (STs) in order to compare their distribution in C, I and E samples. Both clinical and environmental samples harbored prevalent and widely distributed ESBL genes, i.e. blaSHV, blaOXA, blaTEM and blaCTX M. The most prevalent carbapenemase genes were blaNDM-1, blaOXA-48 and blaKPC-2. They were found in all types of isolates, while blaOXA-162, a rare blaOXA-48 variant, was found exclusively in water samples. A higher diversity of carbapenemases genes was seen in wastewater isolates. The aminoglycoside modifying enzymes (AME) genes found in all types of samples were aac(6’), ant(2’’)Ia, aph(3’), aaD, aac(3) and aph(6). Quinolone resistance gene qnrS1 and the multi-drug resistance oqxA/B pump gene were found in all samples, while qnrD and qnrB were associated to aquatic isolates. The antiseptics resistance gene qacEdelta1 was found in all samples, while qacE was detected exclusively in the clinical ones. Trimethroprim-sulfamethoxazole (dfrA, sul1 and sul2), tetracyclines (tetA and tetD) and fosfomycin (fosA6, known to be located on a transpozon) resistance genes were found in all samples, while for choramphenicol and macrolides some ARGs were detected in all samples (catA1 and catB3 / mphA), while other (catA2, cmIA5 and aac(6’)Ib / mphE and msrE) only in wastewater samples. The rifampin resistance genes arr2 and 3 (both carried by class I integrons) were detected only in water samples. The highly prevalent ARGs preferentially associating with aquatic versus clinical samples could ascribe potential markers for the aquatic (blaSHV-145, qacEdelta1, sul1, aadA1, aadA2) and clinical (blaOXA-1, blaSHV-106,blaTEM-150, aac(3)Iia, dfrA14, oqxA10; oqxB17,catB3, tetD) reservoirs of AR. Moreover, some ARGs (oqxA10; blaSHV-145; blaSHV-100, aac(6’)Il, aph(3’) VI, armA, arr2, cmlA5, blaCMY-4, mphE, msrE, oqxB13, blaOXA-10) showing decreased prevalence in influent versus effluent wastewater samples could be used as markers for the efficiency of the WWTPs in eliminating AR bacteria and ARGs. The highest number of virulence genes (75) was recorded for the I samples, while for E and C samples it was reduced to half. The most prevalent belong to three functional groups: adherence (fim genes), iron acquisition (ent, fep, fyu, irp and ybt genes) and the secretion system (omp genes). However, none of the genes associated with hypervirulent K. pneumoniae have been found. A total of 14 STs were identified. The most prevalent clones were ST101, ST219 in clinical samples and ST258, ST395 in aquatic isolates. These STs were also the most frequently associated with integrons. ST45 and ST485 were exclusively associated with I samples, ST11, ST35, ST364 with E and ST1564 with C samples. The less frequent ST17 and ST307 aquatic isolates harbored blaOXA-162, which was co-expressed in our strains with blaCTX-M-15 and blaOXA-1. © 2020 Surleac et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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The Tile Assembly Model, and its many variants, is one of the most fundamental algorithmic assembly formalism within DNA nanotechnology. Most of the research in this field is focused on the complexity of assembling different shapes and patterns. In many cases, the assembly process is intrinsically deterministic and the final product is unique, while the assembly process might evolve through several possible assembly strategies. In this study we consider the controlled assembly of one dimensional tile structures according to predefined assembly graphs. We provide algorithmic approaches for developing such controlled assembly protocols, using the signal-passing Tile Assembly Model, as well as probabilistic approaches for investigating the assembly of such tile-based one-dimensional structures. As a byproduct, we build a generalized TAS (tile assembly system) which generate specific non-local non-associative algebraic computations and we assamble n x n squares using only one tile, which is a better efficiency compared to the staged assembly model. (C) 2019 The Authors. Published by Elsevier B.V.

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Investigation of the lake systems can provide a variety of information that can lead to the development of general concepts about how lakes function and respond to environmental changes. The purpose of this study is to assess the current classification of therapeutic lakes based on supervised learning methods applied to several biochemical characteristics of such lakes. In order to classify the therapeutic lakes in a separate class, a dataset consisting of 45 observations from 9 different basins and from three different altitude categories was analysed using clustering and classification methods.

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The TAM (Model Tile Assembly Model) is a mathematical paradigm for modeling DNA self-assembling according to various given shapes, using DNA-tiles (rectangular shape) with sticky ends on each of the four edges that bound together on various shapes desired by the researcher. Although there are various models in the literature, the focus in this manuscript is on a rule based model, specifically the authors present an overview of the one-dimensional hierarchical self-assembly model of DNA tiles. The authors also present the evolution of number of tiles in partial assemblies, the average assembly size and of the number of partial assemblies of sizes 2 through 10 over the total running time. All simulations were run using the NFSim simulator on a preset period of time.

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This paper presents the latest trends and approaches used for constructing nanoscale structures of 2D objects through DNA folding based on the DNA origami technology developed by Rothemund. The Rothemund method has been used in the construction of various shapes, such as the development of the nanoscale structure for the United States map. Following the steps of Rothemund's technique, we simulate the construction of the Romanian map nanoscale 2D structure, embedding the number 100 into it.

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Screening of 1,000-years old ice layers from the perennial ice block of Scarisoara Ice Cave (NW Romania) revealed the presence of fungal communities. Using culture-dependent methods and molecular techniques based on DGGE fingerprinting of 18S rRNA gene fragments and sequencing, we identified 50 cultured and 14 uncultured fungi in presently-forming, 400 and 900 years old ice layers, corresponding to 28 distinct operational taxonomic units (OTUs). The dominant ice-contained fungal OTUs were related to Ascomycota, Basidiomycota and Cryptomycota phyla. Representatives of Mucoromycota and Chytridiomycota were also isolated from recent and 400 years old ice samples. The cryophilic Mrakia stokesii was the most abundant fungal species found in the cave ice samples of all prospected ages, alongside other cryophilic fungi also identified in various glacial environments. Ice deposits formed during the Little Ice Age (dated between AD 1,250 and 1,850) appeared to have a higher fungal diversity than the ice layer formed during the Medieval Warm Period (prior to AD 1,250). A more complex fungal community adapted to low temperatures was obtained from all analyzed ice layers when cultivated at 4 degrees C as compared to 15 degrees C, suggesting the dominance of cold-adapted fungi in this glacial habitat. The fungal distribution in the analyzed cave ice layers revealed the presence of unique OTUs in different aged-formed ice deposits, as a first hint for putative further identification of fungal biomarkers for climate variations in this icy habitat. This is the first report on fungi from a rock-hosted cave ice block.

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Ice entrenched microcosm represents a vast reservoir of novel species and a proxy for past climate reconstitution. Among glacial ecosystems, ice caves represent one of the scarcely investigated frozen habitats. To characterize the microbial diversity of perennial ice from karst ecosystems, Roche 454 sequencing of 16S rRNA gene amplicons from the underground ice block of Scarisoara Ice Cave (Romania) was applied. The temporal distribution of bacterial and archaeal community structures from newly formed, 400, and 900 years old ice layers was surveyed and analyzed in relation with the age and geochemical composition of the ice substrate. The microbial content of cave ice layers varied from 3.3 10(4) up to 7.5 10(5) cells mL(-1), with 59-78% viability. Pyrosequencing generated 273,102 reads for the five triplicate ice samples, which corresponded to 3,464 operational taxonomic units (OTUs). The distribution of the bacterial phyla in the perennial cave ice varied with age, organic content, and light exposure. Proteobacteria dominated the 1 and 900 years old organic rich ice deposits, while Actinobacteria was mostly found in 900 years old ice strata, and Firmicutes was best represented in 400 years old ice. Cyanobacteria and Chlorobi representatives were identified mainly from the ice block surface samples exposed to sunlight. Archaea was observed only in older ice strata, with a high incidence of Crenarchaeota and Thaumarchaeaota in the 400 years old ice, while Euryarchaeota dominated the 900 years old ice layers, with Methanomicrobia representing the predominant taxa. A large percentage (55.7%) of 16S rRNA gene amplicons corresponded to unidentified OTUs at genus or higher taxa levels, suggesting a greater undiscovered bacterial diversity in this glacial underground habitat. The prokaryotes distribution across the cave ice block revealed the presence of 99 phylotypes specific for different ice layers, in addition to the shared microbial community. Ice geochemistry represented an important factor that explained the microbial taxa distribution in the cave ice block, while the total organic carbon content had a direct impact on the cell density of the ice microcosm. Both bacterial and archaeal community structures appeared to be affected by climate variations during the ice formation, highlighting the cave ice microbiome as a source of putative paleoclimatic biomarkers. This report constitutes the first high-throughput sequencing study of the cave ice microbiome and its distribution across the perennial underground glacier of an alpine ice cave.

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Screening of 1,000-years old ice layers from the perennial ice block of Scǎrişoara Ice Cave (NW Romania) revealed the presence of fungal communities. Using culture-dependent methods and molecular techniques based on DGGE fingerprinting of 18S rRNA gene fragments and sequencing, we identified 50 cultured and 14 uncultured fungi in presently-forming, 400 and 900 years old ice layers, corresponding to 28 distinct operational taxonomic units (OTUs). The dominant ice-contained fungal OTUs were related to Ascomycota, Basidiomycota and Cryptomycota phyla. Representatives of Mucoromycota and Chytridiomycota were also isolated from recent and 400 years old ice samples. The cryophilic Mrakia stokesii was the most abundant fungal species found in the cave ice samples of all prospected ages, alongside other cryophilic fungi also identified in various glacial environments. Ice deposits formed during the Little Ice Age (dated between AD 1,250 and 1,850) appeared to have a higher fungal diversity than the ice layer formed during the Medieval Warm Period (prior to AD 1,250). A more complex fungal community adapted to low temperatures was obtained from all analyzed ice layers when cultivated at 4 °C as compared to 15 °C, suggesting the dominance of cold-adapted fungi in this glacial habitat. The fungal distribution in the analyzed cave ice layers revealed the presence of unique OTUs in different aged-formed ice deposits, as a first hint for putative further identification of fungal biomarkers for climate variations in this icy habitat. This is the first report on fungi from a rock-hosted cave ice block. © 2018 The Author(s).

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In the last few decades, due to the global industrialization and population expansion the level of pollutants has largely increased, being one of the main environmental problems faced worldwide. Consequently, phytoremediation research had been gathering more and more interest. A study of data selected from Scopus is presented by the authors, identifying trends in publication number, collaboration and impact. The trends are identified at the global level followed by a discussion of how Romania performs with respect to the identified trends. When evaluating the research produced by the Romanian institutions, two characteristics are considered: phytoremediation potential of the plants in the Danube Delta and the coastal region and the phytoremediation research temporal evolution and international impact. The paper presents a quantitative analysis without adding variable weights to citations and publications based on the publications ranks offering an overview of the national research status in this research area.

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The paper deals with the cellulase-mediated hydrolysis applied to several residual biomass resulting after extraction of biologically active principles from three medicinal plants: Melissa officinalis L., Melilotus officinalis L., Viola tricolor L. The yield of hydrolysis to hexoses was determined based on the amount of free glucose in the reaction mixture identified by spectrophotometric analysis, and total conversion of biomass residues was calculated. It was found that biomass resulted from Viola tricolor L. exhibits best glucose amount (10.35%) after 1 h of enzymatic hydrolysis, while that sourced from Melissa officinalis L. gives the highest total conversion of solid biomass (82.13%).

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The paper present histo-anatomic aspects, recorded in seedlings of maize (Zea mays L.) grown under the influence of spruce bark polyphenolic extract (SBPE). Spruce barck polyphenolic extract was characterized in terms of the total content of polyphenols, tannins, flavonols, flavonoids and anthocyanins in a previous paper. It is known that, natural polyphenols are essential compounds in stimulating plant growth and development. The aim of this study was to identify the internal structure changes in maize seedlings, under the influence of SBPE, applied in two concentrations (SB1 - 190 mg GAE / L and SB2 - 130 mg GAE / L). It was observed, compared with the control, that SBPE determines a better development of the absorbents bristle and increase leading bundles number and central vessels metaxylem. SBPE in lower concentrations has positive effects that can be attributed to antioxidant properties. The results recorded in this study open new pathways of research in the field.

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The Danube River-Danube Delta-Black Sea (DBS) region has witnessed major political, social and economic changes during the past three decades, which have profoundly affected the riverine, coastal and marine systems, their water management situation and the development of related research programmes. We reviewed the research activities in the DBS system of the past twenty years to determine the main funding bodies and to assess key research areas and how they varied over time and geographic region. As data basis we used a metadatabase filled with 478 projects addressing environmental and water management issues in the Danube River Basin, covering also the Danube Delta and the north-western Black Sea. As overall outcome extensive research efforts in the field of water management could be proven for the past two decades, despite the tumultuous times of political and economic transformations. One of the main findings was that EU funded projects played a key role for the development of transboundary research collaboration and were also the scientifically most productive one's. Historically, nutrient pollution was the main problem addressed, shifting to pollution in a broader sense and hydromorphological alterations in recent years. The newly arising challenges of climate change impacts and sediment management became important research questions in the last years, too. Most research was performed in the thematic field of navigation, followed by restoration and biodiversity issues. To meet all of the already

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The Danube River–Danube Delta–Black Sea (DBS) region has witnessed major political, social and economic changes during the past three decades, which have profoundly affected the riverine, coastal and marine systems, their water management situation and the development of related research programmes. We reviewed the research activities in the DBS system of the past twenty years to determine the main funding bodies and to assess key research areas and how they varied over time and geographic region. As data basis we used a metadatabase filled with 478 projects addressing environmental and water management issues in the Danube River Basin, covering also the Danube Delta and the north-western Black Sea. As overall outcome extensive research efforts in the field of water management could be proven for the past two decades, despite the tumultuous times of political and economic transformations. One of the main findings was that EU funded projects played a key role for the development of transboundary research collaboration and were also the scientifically most productive one's. Historically, nutrient pollution was the main problem addressed, shifting to pollution in a broader sense and hydromorphological alterations in recent years. The newly arising challenges of climate change impacts and sediment management became important research questions in the last years, too. Most research was performed in the thematic field of navigation, followed by restoration and biodiversity issues. To meet all of the already identified and newly emerging challenges in the DBS System, cross-border and integrated (river-delta-sea) research activities are of major importance and have to be further promoted. We thus suggest drawing up a regional DBS Research Agenda linked to key challenges in water management to strengthen research collaboration and advance targeted scientific projects, an approach fostering also the scientific capacity in the region. © 2015 Elsevier B.V.

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According to European Commission reports published between 2010 - 2013, the development of European economy depends crucially on access to critical raw materials. Following the analysis performed by experts at European level, in 2011 was compiled and published a list of 14 critical raw materials, the so-called EU-14. In 2014 the list was updated with several new elements and one element (tantalum), was withdrawn from the list. The current list, being renamed EU-20, covers 20 critical raw materials including several high tech critical metals. Traditional mine exploitations are concentrated on using the deposits of ore extracted and processed by conventional techniques. The efficiency of metal recovery was variable over time and as a result, a significant amount of metal was discarded, most concentrations exceeding the current minimal permissible threshold. On the other hand, it is necessary the recovery of recyclable waste for reducing the risk of shortage of high tech critical metals. Therefore, it is necessary to develop new technologies for obtaining high tech critical metals, which is applicable to both primary and secondary sources of raw materials. Recovery of high-tech critical metals by processing ore, tailings or mine wastes, and recyclable materials can be successfully done with help of consortia or individual isolates of microorganisms, bacteria or fungi. Microorganisms interact with metals thus altering their physical and chemical condition. Isolation of individual strains and identification of microbial consortia that can be used in the design and development of effective biotechnological processes for the extraction of high tech critical metals is a current challenge of the scientific research in Europe. © 2016 Vasile Goldis University Press.

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The regional bioeconomic potential takes into consideration many sectors of the economy that use renewable biological resources. The plants are one of the main resources for the most sectors - food, health, environment, materials and energy. The assessment of bioeconomic potential involves the modeling of the biological and economical aspects. Establishing the indicators involved in the study of this aspect requires a “what we have and what we need” analysis. We started with the development of a database of plants to facilitate an easy access to information for all interested scientist from related research fields. © 2016 Vasile Goldis University Press.

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The paper summarises the results of the 478 projects from the FP7-ENVIRONMENT proiect, DANube macroregion: Capacity building and Excellence in River Systems (basin, delta and sea) DANCERS database considering different attributes recorded for these projects. An analysis is performed on the recorded data and the significant characteristics are presented. The analysis performs a research output assessment in order to quantify the success of the projects. Trends are identified not only by thematic area and region of implementation, but also by coordinating country region and level of financing. A need to homogenising the level of financing as well as the level of funding efforts among the EU Strategy for the Danube Region (EUSDR) countries and not only, transpires at the end of the analysis. A concentrated effort towards inter and intra disciplinary collaboration as well as within and between institutional collaboration and knowledge sharing is also apparent.

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Transfusion medicine relies on the possibility of storing red blood cells (RBCs) for a prolonged period of time. A better understanding of the nature of changes in stored RBCs may provide new strategies to improve the balance of benefits and transfusion's risks. In the light of recent results on the mechanism of programmed cell death of RBCs (erythroptosis), where numerous cellular changes are common hallmarks of the apoptotic phenomenon, we have developed new flow cytometric criteria to evaluate the viability of stored erythrocytes. In order to evaluate the possible benefic effect of low-intensity laser irradiation for increasing the blood preservation period, we irradiated the whole blood from healthy donors in specially designed bags for research purposes by MacoPharma Company containing SAGM medium, and conserved it for 3 weeks. LED (Light emitting diode) (lambda=465nm, P=60mW, P/S=100mW/cm(2)) and laser diode (lambda=660nm, P=30mW, P/S= 50mW/cm(2)) in continuous wave have been used as sources of irradiation. Doses were ranged between 0 and 2J/cm(3). After that period RBCs were analyzed by flow cytometry for morphological changes (FSC/SSC), apoptosis/necrosis analysis (FITC-annexin-V labeling/PI) and viability using an original Calcein-AM method on a FACScan cytometer using CellQuest Pro software for acquisition and analysis. Complementary, the RBCs integrity was analyzed by scanning electron microscopy (SEM). Morphological changes assessment of red blood cells by flow cytometry and scanning electron microscopy showed a good preservation of the discoidal shape for the irradiated cells compared with control.

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In this work we describe a new segmentation technique for the Affymetrix microarray images. We prove that our method can offer better predictions on the gene levels as opposed to the standard Affymetrix segmentation implemented in the Affymetrix GeneChip Operating Software (GCOS). To check the accuracy and show the benefits of the new segmentation method we use a previously implemented methodology to simulate microarray images with realistic features. Using such an artificial image provides us with the actual levels for each spot and each gene investigated in the microarray. Using this information we then proceed to segment the same image twice (with GCOS and our new method). The two segmentations will produce two sets of gene levels that are then compared to the known gene levels (known since the moment of generating the artificial image). Using this methodology we are able to show statistically (using 50 replicates of the same steps of generating the image, segmenting, comparing the results) that in some cases our new method greatly outperforms the GCOS implemented segmentation method, while in the rest of the cases performs in similar fashion. (C) 2015 Elsevier B.V. All rights reserved.

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Composite gels based on collagen and eucalyptus oil were prepared in order to be used in dentistry. The main components such as beta-Cymene (32.1%), Eucalyptol (36.59%), Cryptone (4.35%) and Spathulenol (3.82%) of Eucalyptus camadulensis Dehnh. essential oil were determined by GC-MS. The eucalyptus oil dissolved in different concentration of ethanol (4 and 8% v/v) was embedded in collagen gels (0.4 and 0.8% w/w) and crosslinked with tannic acid (0.2 and 0.4% w/w). The stability of gels was depending on collagen concentration, only the gels with 0.8% of collagen being stable. The rheological analysis of composite gels showed that formulation containing a combination of ethanol and tannic acid at the ratio of 8% v/v: 0.4% w/w resulted in the highest consistency index and yield stress. All the 0.8% collagen gels exhibited a suitable pseudoplastic behaviour and could be recommended for an adequate administration in the oral cavity.

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The paper refers to a bioactive matrix in membrane shape, with a thickness of 0.5 - 1.0 mm, and consisting of gelatine, chitosan, and α-chymotrypsin, with or without addition of glutaraldehyde as a cross-linking agent. The bioproduct is a biomaterial with biomedical applications as biocompatible membrane with anti-inflammatory action due to coupling of the proteolytic enzyme. Bioproducts have complex implications in the treatment of traumatized tissue, accelerating the pus cleansing of infected wound, blood clots and other cellular debris, secretions liquefaction and cleansing of necrotic tissue, and also stimulating tissue regeneration and a faster healing of open infected wounds. In order to establish the influence of cross- linking process and membrane’s thickness on mechanical properties, ultrastructural studies were carried out on obtained biomaterials by electron microscopy analysis. To demonstrate the biological qualities of new enzymatic biomaterials, the membrane’s biological effect was analyzed in a cell culture model (mouse fibroblasts NCTC). © 2014 Vasile Goldis University Press.

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The use of nanoparticles for biological and medical applications has rapidly increased and the potential for human and ecological toxicity is a growing area of investigation. For assessing cytotoxicity of nanoparticles we developed a new experimental cell system based on the use of nucleated erythrocytes (RBCs) from fish and batrachians, which are directly exposed to pollutants or nanoparticles absorbed by different ways and we have evaluated the toxic effects by flow cytometric analysis. The two modes of cell death (apoptosis and necrosis) differ fundamentally in their morphology, biochemistry and biological relevance. We and others have recently shown that programmed cell death of nucleated erythrocytes is related to an apoptotic mechanism. The toxicological analysis were performed comparatively on porphyrin base or metalloporphyrin and for each porphyrin bare derivative on two correspondent porphyrin-silica-hybrid nanomaterials obtained by one step acid catalysis and by two steps acid-base catalysis. To evaluate cell-nanoparticles interactions, nucleated RBCs were exposed to different concentrations of nanocomposites and analyzed by flow cytometry, after 24h incubation endpoints for morphological changes (FSC/SSC), apoptosis/necrosis analysis (FITC-annexin-V labeling/PI) and viability (using calcein-AM method). The investigation showed that the type of cellular death of nucleated erythrocytes is related to an apoptotic mechanism and that flow cytometric analysis of nucleated RBCs viability and cell death discrimination could provide a rapid and accurate analytical tool for evaluating in vitro the biological responses towards of nanoparticles for environmental protection. Nucleated erythrocytes can be a new experimental cellular model easy to use, with no costs for culture and for maintaining in the culture. The results reported in the present study indicate that our new flow cytometric protocols can be used to create dose-response curves which allow us to determine EC50 for toxicity or ecotoxicity tests. It is also generally applicable for identifying harmful effects associated with general antropic impact for the aquatic environment and for its biomonitoring.

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Polycythemia vera (PV) is characterized by an absolute increase in the red blood cell mass, but the mechanisms are not completely understood. In this study, we identified by flow cytometric analysis morphological forms that deviate from the classical discoid shape, who had level. Measurement of glycoconjugate sialylation using lectines demonstrates but which is close to normal after treatment and an increased percentage of cells with active caspase-8 and -3 compared to normal RBCs, showing that the organism tries to restore the apoptotic mechanism for maintaining the normal hematocrit. may contribute elucidation of the mechanism this diseasea more viability determined by Calcein-AM method and a normal phosphatidylserine exposure a low degree of sialilation of membrane glycoconjugates of Polycythemia vera RBCs Our observations for understanding the survival of RBCs in Polycythemia vera and may also participate in elucidation of the mechanism this diseasein pathogenesis of this disease. © 2011 Vasile Goldis University Press.

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The human activities have a negative impact to the environment, consisting in the water contamination with toxic products, heavy metals or with xenobiotic substances. Manufactured nanomaterials (nanoparticles, nanotubes, nanosheets and nanowires) have recent applications in drug delivery, medical devices, cosmetics, chemical catalysts, optoelectronics, electronics and magnetics. Some nanomaterials have been found to be toxic to humans and other organisms either upon contact or after persistent environmental exposure. In present, the measurements of the pollution degree are made with two methods: phisyco-chemical methods and ecotoxicological test (bioassay or environmental biosensors). Our results indicate that flow cytometric analysis of nucleated red blood cells viability using calcein-AM and cell death discrimination could provide a rapid and accurate experimental cellular model for effectively screening and evaluating biological responses for in vitro nanotoxicology and can be used in ecotoxicology as bioassays for the ecological monitoring of aquatic environment. In the some time, our results indicate that the use of nucleated erythocytes could be potentially useful for the development of rapid and low cost safety tests to assess fisheries product quality. © 2011 Vasile Goldis University Press.