Oxidative Polymerization of 3,4-Dihydroxybenzylamine─The Lower Homolog of Dopamine.

Polydopamine (PDA) formed by oxidative polymerization of dopamine has attracted wide interest because of its unique properties, in particular its strong adhesion to almost all types of surfaces. 3,4-Dihydroxybenzylamine (DHBA) as the lower homolog of PDA also contains a catechol unit and an amino group and thus can be expected to exhibit a similar adhesion and reaction behavior. In fact, autoxidation of DHBA with air in 2-amino-2-hydroxymethyl-propane-1,3-diol (Tris) buffer gives rise to deeply colored oligomer/polymer products (poly(3,4-dihydroxybenzylamine) (PDHBA)) that strongly adhere to several surfaces. Here, the material is characterized by solid-state NMR spectroscopy, Fourier transform infrared (FTIR) spectroscopy, X-ray photoelectron spectroscopy (XPS), electron spin resonance (ESR) spectroscopy, mass spectrometry, and atomic force microscopy (AFM). Reaction pathways were rationalized taking into consideration the analytical results that show similarity to PDA chemistry, but also considering differences, leading to a more complex reaction behavior and thus to new structures not found in PDA.

Bevacizumab Treatment for Metastatic Colorectal Cancer in Real-World Clinical Practice.

Background and Objectives: Colorectal cancer (CRC) is a leading cause of cancer-related mortality and morbidity worldwide. Bevacizumab was approved for the treatment of metastatic colorectal cancer (mCRC) based on favorable benefit-risk assessments from randomized controlled trials, but evidence on its use in the real-world setting is limited. The aim of the current study is to evaluate the outcomes and safety profile of bevacizumab in mCRC in a real-world setting in Romania. Patients and Methods: This was an observational, retrospective, multicentric, cohort study conducted in Romania that included patients with mCRC treated with bevacizumab as part of routine clinical practice. Study endpoints were progression-free survival, overall survival, adverse events, and patterns of bevacizumab use. Results: A total of 554 patients were included in the study between January 2008 and December 2018. A total of 392 patients (71%) received bevacizumab in the first line and 162 patients (29%) in the second line. Bevacizumab was mostly combined with a capecitabine/oxaliplatin chemotherapy regimen (31.6%). The median PFS for patients treated with bevacizumab was 8.4 months (interquartile range [IQR], 4.7-15.1 months) in the first line and 6.6 months (IQR, 3.8-12.3 months) in the second line. The median OS was 17.7 months (IQR, 9.3-30.6 months) in the first line and 13.5 months (IQR, 6.7-25.2 months) in the second line. Primary tumor resection was associated with a longer PFS and OS. The safety profile of bevacizumab combined with chemotherapy was similar to other observational studies in mCRC. Conclusions: The safety profile of bevacizumab was generally as expected. Although the PFS was generally similar to that reported in other studies, the OS was shorter, probably due to the less frequent use of bevacizumab after disease progression and the baseline patient characteristics. Patients with mCRC treated with bevacizumab who underwent resection of the primary tumor had a higher OS compared to patients with an unresected primary tumor.

Diversity, distribution and organic substrates preferences of microbial communities of a low anthropic activity cave in North-Western Romania.

Introduction: Karst caves are characterized by relatively constant temperature, lack of light, high humidity, and low nutrients availability. The diversity and functionality of the microorganisms dwelling in caves micro-habitats are yet underexplored. Therefore, in-depth investigations of these ecosystems aid in enlarging our understanding of the microbial interactions and microbially driven biogeochemical cycles. Here, we aimed at evaluating the diversity, abundance, distribution, and organic substrate preferences of microbial communities from Peștera cu Apa din Valea Leșului (Leșu Cave) located in the Apuseni Mountains (North-Western Romania). Materials and Methods: To achieve this goal, we employed 16S rRNA gene amplicon sequencing and community-level physiological profiling (CLPP) paralleled by the assessment of environmental parameters of cave sediments and water. Results and Discussion: Pseudomonadota (synonym Proteobacteria) was the most prevalent phylum detected across all samples whereas the abundance detected at order level varied among sites and between water and sediment samples. Despite the general similarity at the phylum-level in Leșu Cave across the sampled area, the results obtained in this study suggest that specific sites drive bacterial community at the order-level, perhaps sustaining the enrichment of unique bacterial populations due to microenvironmental conditions. For most of the dominant orders the distribution pattern showed a positive correlation with C-sources such as putrescine, γ-amino butyric acid, and D-malic acid, while particular cases were positively correlated with polymers (Tween 40, Tween 80 and α-cyclodextrin), carbohydrates (α-D-lactose, i-erythritol, D-mannitol) and most of the carboxylic and ketonic acids. Physicochemical analysis reveals that sediments are geochemically distinct, with increased concentration of Ca, Fe, Al, Mg, Na and K, whereas water showed low nitrate concentration. Our PCA indicated the clustering of different dominant orders with Mg, As, P, Fe, and Cr. This information serves as a starting point for further studies in elucidating the links between the taxonomic and functional diversity of subterranean microbial communities.

Competition-cooperation in the chemoautotrophic ecosystem of Movile Cave: first metagenomic approach on sediments.

BACKGROUND: Movile Cave (SE Romania) is a chemoautotrophically-based ecosystem fed by hydrogen sulfide-rich groundwater serving as a primary energy source analogous to the deep-sea hydrothermal ecosystems. Our current understanding of Movile Cave microbiology has been confined to the sulfidic water and its proximity, as most studies focused on the water-floating microbial mat and planktonic accumulations likely acting as the primary production powerhouse of this unique subterranean ecosystem. By employing comprehensive genomic-resolved metagenomics, we questioned the spatial variation, chemoautotrophic abilities, ecological interactions and trophic roles of Movile Cave's microbiome thriving beyond the sulfidic-rich water. RESULTS: A customized bioinformatics pipeline led to the recovery of 106 high-quality metagenome-assembled genomes from 7 cave sediment metagenomes. Assemblies' taxonomy spanned 19 bacterial and three archaeal phyla with Acidobacteriota, Chloroflexota, Proteobacteria, Planctomycetota, Ca. Patescibacteria, Thermoproteota, Methylomirabilota, and Ca. Zixibacteria as prevalent phyla. Functional gene analyses predicted the presence of CO2 fixation, methanotrophy, sulfur and ammonia oxidation in the explored sediments. Species Metabolic Coupling Analysis of metagenome-scale metabolic models revealed the highest competition-cooperation interactions in the sediments collected away from the water. Simulated metabolic interactions indicated autotrophs and methanotrophs as major donors of metabolites in the sediment communities. Cross-feeding dependencies were assumed only towards 'currency' molecules and inorganic compounds (O2, PO43-, H+, Fe2+, Cu2+) in the water proximity sediment, whereas hydrogen sulfide and methanol were assumedly traded exclusively among distant gallery communities. CONCLUSIONS: These findings suggest that the primary production potential of Movile Cave expands way beyond its hydrothermal waters, enhancing our understanding of the functioning and ecological interactions within chemolithoautotrophically-based subterranean ecosystems.

3D silver metallized nanotrenches fabricated by nanoimprint lithography as flexible SERS detection platform.

We report the development of highly sensitive substrates with great potential as Surface-enhanced Raman scattering (SERS) spectroscopy detection platforms, consisting of nanoimprint lithography (NIL) fabricated nanotrenches in plastic and covered by nanostructured silver (Ag) films with thicknesses in the 10-100 nm range deposited by direct current (DC) sputtering. The Ag film thickness was increased by using sequential deposition times and its contribution to the obtained enhancement factor was determined. The morphological and structural properties of the metalized nanotrenches were assessed by scanning electron microscopy (SEM) and atomic force microscopy (AFM) techniques. Crystal violet (CV) was used as analyte to test the SERS activity of the substrates prepared with or without the nanoimprinted pattern. Our original approach was to determine the resulted SERS enhancement from the synergy of three key aspects: the Ag metallization of cheap, flexible substrates, the effect of increasing the Ag film thickness and the periodic nanotrenches imprinted by NIL as substrate. We found a dramatical contribution in the SERS signal of the periodical Ag nanopattern in comparison to the Ag film quantified by a calculated enhancement factor (EF) up to 107 in case of the SERS detection platform with a 25 nm Ag layer on top of the periodic nanotrenches. The contribution of plasmonic nanostructures contained in the Ag films as well as the contribution of the periodical nanopatterned trenches was assessed, as a cumulative effect to the first contribution. This substrate showed a considerably lower limit of detection (LOD) for SERS, down to 10 pM, much better uniformity as well as more reproducible signals in comparison with the other thicknesses of the metallic film.

Gold-based nanostructured platforms for oxytetracycline detection from milk by a "signal-on" aptasensing approach.

Several gold platforms of different morphologies were investigated in the elaboration of a new aptasensor for oxytetracycline. Au-nanostructures were electrochemically synthesized from solutions of different concentrations of HAuCl4 in different media by chronoamperometry, multipulse amperometry, and chronopotentiometry, respectively at carbon-based screen-printed electrodes (C-SPE). The nano-/micro-scale morphologies of the patterned surfaces and elemental composition were examined by scanning electron microscopy, atomic force microscopy, and X-ray photoelectron spectroscopy, respectively. The electrochemical properties of the obtained gold nanostructured platforms (AuNSs|C-SPE) were investigated to achieve optimal aptamer coverage. The results showed that the aptasensor developed using the platform with thistle-like AuNSs exhibited the highest conductivity in terms of ferrocene signal and the largest effective area. Under optimal conditions, a linear range from 5.0 × 10-8 M to 1.2 × 10-6 M, with a limit of detection (LOD) of 8.7 × 10-9 M OXT were obtained, which is about 20 times lower than the EU regulations for OXT residues in milk. The electrochemical aptasensor was able to discriminate other antibacterial agents, such as amoxicillin, ampicillin, gentamicin, tetracycline, and vancomycin and was successfully applied in milk samples. This "signal-on" aptasensing approach provides a simple and cost-effective disposable sensor that could be easily applied for the on-site determination of antibiotics.

Sequential aqueous two-phase system for simultaneous purification of cyanobacterial phycobiliproteins.

A logical framework was used for designing a top-down strategy for cyanobacterial phycobiliprotein purification. The purification scheme is based on the non-chromatographic technique, known as aqueous two-phase system. The scheme was optimized at every stage to enhance the recovery yield with the highest purity. We tested this strategy on four cyanobacteria, two containing only phycocyanin and allophycocyanin (Arthrospira platensis AICB49, Synechocystis sp. AICB51) and two that have an extra phycobiliprotein, namely phycoerythrin (Fremyella sp. UTEX481, Coelomoron pussilum AICB1012). The results showed that the recovery efficiency of the phycobiliproteins is strongly influenced by the phycobilisome composition. For the first two strains the recovery yield of both phycocyanin and allophycocyanin was >80%, with an analytical purity grade for phycocyanin (>4.2) and a reactive purity grade for allophycocyanin (>2.9). The recovery yield of phycoerythrin was lower but compensating with an increase in purity, 5.2 for Fremyella and 4.5 for C. pussilum.

Biomimetic electrochemical sensor for the highly selective detection of azithromycin in biological samples.

A highly selective and sensitive molecularly imprinted polymer (MIP)-based electrochemical sensor was fabricated for the determination of azithromycin, a broad-spectrum macrolide antibiotic, from various biological samples (urine, tears, plasma). The reversible boronate ester bond-mediated, thin (~75 nm) MIP-based biomimetic recognition layer was electrodeposited in non-aqueous media onto the surface of a glassy carbon electrode (GCE). The surface morphology and the analytical performances of the developed sensor were assessed by scanning electron (SEM) and atomic force microscopy (AFM), electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). By employing an indirect electrochemical detection in the presence of 10 mM ferro/ferricyanide as redox probe, the sensor exhibited a very wide dynamic range (13.33 nM-66.67 µM), with an estimated detection limit in the subnanomolar range (0.85 nM azithromycin). The simple to construct sensor demonstrates reusability and good shelf-life, exhibiting remarkable selectivity over a wide number of structurally related and non-related antibiotics, commonly associated drugs and endogenous compounds.

Impedimetric aptasensor for the label-free and selective detection of Interleukin-6 for colorectal cancer screening.

Despite the fact that cancer research has experienced important advances and remarkable improvement in the curing processes during the last decades, this disease still occupies a leading position among the causes of death worldwide. It has been demonstrated that there is an interconnection between the overexpression of interleukin-6 cytokine and the tumor growth, metastasis, and therapeutic resistance in several types of malignancies. Herein, a highly sensitive and selective aptasensor for quantitative detection of interleukin-6 was developed by using a glassy carbon electrode modified with p-aminobenzoic acid, p-aminothiophenol and gold nanoparticles. A thio-terminated aptamer specific for interleukin-6 was immobilized on the surface of the modified electrode via the formation of gold-sulfur bonds. This DNA oligonucleotide was then used as a detection probe to capture the target protein at the biosensor surface allowing label-free detection by electrochemical impedance spectroscopy. The developed aptasensor showed a good linear response from 5 pgmL-1 to 100 ngmL-1 with a detection limit of 1.6 pgmL-1, within the range of physiological concentration of the protein. The biosensor exhibited high selectivity and has been successfully used to detect interleukin-6 in blood samples collected from patients suffering of colorectal cancer, with excellent recoveries after the addition of known amount of the target protein.

Homogeneity assessment for grass samples used for organically bound tritium proficiency test.

The paper presents homogeneity check for dried grass batch to be used in an inter-comparison exercise. When carrying out an inter-comparison exercise, the organizer must guarantee that the unsatisfactory result will not be imputed to the sample and sample's changeability. Therefore, in an inter-comparison exercise must carry on the homogeneity testing. In our study, the homogeneity was checked for two parameters, the hydrogen percentage (%H) and the organically bound tritium (OBT) activity concentration. Ten subsamples of 50g from bulk material were chosen using a stratified random selection. Two replicates from each package were analyzed for hydrogen content and OBT analysis. The homogeneity parameters were evaluated according to ISO 13528:2015. The two checked properties confirm that the batch of dried grass is sufficiently homogenous.

Characterization and Discrimination of Gram-Positive Bacteria Using Raman Spectroscopy with the Aid of Principal Component Analysis.

Raman scattering and its particular effect, surface-enhanced Raman scattering (SERS), are whole-organism fingerprinting spectroscopic techniques that gain more and more popularity in bacterial detection. In this work, two relevant Gram-positive bacteria species, Lactobacillus casei (L. casei) and Listeria monocytogenes (L. monocytogenes) were characterized based on their Raman and SERS spectral fingerprints. The SERS spectra were used to identify the biochemical structures of the bacterial cell wall. Two synthesis methods of the SERS-active nanomaterials were used and the recorded spectra were analyzed. L. casei and L. monocytogenes were successfully discriminated by applying Principal Component Analysis (PCA) to their specific spectral data.

Detection of Dopamine by a Biomimetic Electrochemical Sensor Based on Polythioaniline-Bridged Gold Nanoparticles.

A new biomimetic electrochemical sensor was developed for the detection of dopamine based on a glassy carbon electrode modified with electrochemically generated gold nanoparticles. The preparation of the polymer is simple and cost-effective, achieving the polymerization of thioaniline and generation of gold nanoparticles in a single step by cyclic voltammetry, in the presence of the target molecule, dopamine. After extraction, the imprinted polymer exhibits high sensitivity and selectivity for dopamine. Moreover, the developed imprinted polymer film allows the fast, direct detection of dopamine without the need of a redox mediator. The formation of a self-assembled monolayer of the monomer prior to electropolymerization ensures the adherence of the film onto the electrode surface, conferring good stability to the sensor (over two weeks). Cyclic voltammetry, electrochemical impedance spectroscopy, atomic force microscopy, scanning electron microscopy, and energy dispersive X-ray spectroscopy were used for the complete characterization of the developed sensor, and differential pulse voltammetry was used for its testing.

Effect of van der Waals interaction on the geometric and electronic properties of DNA nucleosides adsorbed on Cu(111) surface: a DFT study.

The geometrical properties and electronic structure of DNA nucleosides (deoxyadenosine, thymidine, deoxyguanosine, deoxycytidine) adsorbed on a metallic surface of Cu(111) are determined using density functional theory computations. In order to assess the effect of the long-range interaction upon the results of the DFT simulations, we compare the results of a standard GGA exchange-correlation functional with those produced by the newly developed van der Waals exchange-correlation functional. The most striking difference between the two sets of results occurs for the adsorption energies: standard functional predicts values representing about 30% of those obtained when van der Waals interaction is taken into account. The standard GGA functional favors slightly tilted orientation of the DNA bases with respect to the surface, while the inclusion of the van der Waals effect leads to an almost parallel orientation of the bases with the surface. On the other hand, the position of the sugar pucker is less influenced by the type of the exchange-correlation used. According to our studies, in the presence of long-range interactions, the molecule-surface charge transfer is qualitatively affected. Standard functional predicts a decrease of the electronic population of the adsorbate, while the presence of long-range interaction leads to an opposite effect.

Binding effects of Mn²âº and Zn²âº ions on the vibrational properties of guanine-cytosine base pairs in the Watson-Crick and Hoogsteen configurations.

The binding effects of Mn²âº and Zn²âº ions on the vibrational properties of guanine-cytosine base pairs have been performed using density functional theory investigations. The calculations were carried out on Watson-Crick and Hoogsteen configurations of the base pairs. We have found, that in Watson-Crick configuration, the metal is coordinated to N7 atom of guanine while, in the case of Hoogsteen configuration, the coordination is at N3 atom of guanine. We have pointed out the vibrational bands that can be used to detect the presence of metallic ions in the Watson-Crick and Hoogsteen structures. Our results show that the vibrational amplitudes of metallic atoms are strong for wavenumbers lower than 600 cm⁻¹. Also, we predict that the distinction between Watson-Crick and Hoogsteen configurations can be seen around 85, 170 and 310 cm⁻¹.

Localization and anharmonicity of the vibrational modes for GC Watson-Crick and Hoogsteen base pairs.

We present an ab initio study of the vibrational properties of cytosine and guanine in the Watson-Crick and Hoogsteen base pair configurations. The results are obtained by using two different implementations of the DFT method. We assign the vibrational frequencies to cytosine or to guanine using the vibrational density of states. Next, we investigate the importance of anharmonic corrections for the vibrational modes. In particular, the unusual anharmonic effect of the H(+) vibration in the case of the Hoogsteen base pair configuration is discussed.

Dependence of the size of a protein-SDS complex on detergent and Na+ concentrations.

Sodium dodecyl sulfate (SDS) micelles provide ideal mimetic media for high-resolution NMR studies of membrane proteins and proteins or peptides interacting with micellar aggregates. (15)N NMR relaxation of the backbone amides of a protein-SDS complex has been measured under different experimental conditions. The rotational diffusion time of this complex has been found highly sensitive to detergent and NaCl concentrations. A comparison with calculated rotational diffusion times of protein-free SDS micelles under the same conditions suggests that the size of both aggregates must follow a similar functional dependence on detergent/NaCl concentration.

A study of the anharmonic effects on the vibrational spectra of a realistic retinal chromophore model.

Ab initio and vibrational self-consistent field (VSCF) computations are used to investigate the vibrational normal coordinates of the protonated Schiff base (PSB) 4-cis-gamma,eta-dimethyl-C9H9 NH2+. The ground and the first excited states are investigated. Both harmonic and anharmonic frequencies for the first three overtones of the ground and first excited states are reported. Special attention is payed to the discussion of the normal coordinates modes that involve the central C=C bond which plays a significant role in the isomerization process.