Accurate determination of molar mass distribution for disperse cellulose samples has proved to be a challenging task. While size-exclusion chromatography coupled to multi-angle light scattering ...(MALS) and differential refractive index (DRI) detectors has become the most commonly used method for molar mass determination of celluloses, this technique suffers low sensitivity at the low-molar mass range. As discussed here, the universal method for accurate molar mass distribution analysis of cellulose samples not exists and thus thorough understanding on the differences of the various methodological approaches is important. In this study, the focus is in the accurate determination of the low-molar mass fraction. The results obtained by combining the two calibration strategies, MALS/DRI for polymeric region of a cellulose sample and conventional calibration for oligomeric region, was compared to the results obtained using only MALS/DRI (with extrapolation of the curve where signal-to-noise of MALS is low). For birch pulp sample, the results from the two approaches were comparable; it should be highlighted, however, that MALS/DRI slightly overestimates the molar masses at the low-molar-mass region.
•Size-exclusion chromatography (SEC) of metal nanoparticles (NPs) and quantum dots (QDs) is reviewed.•Problems associated with SEC analysis of NPs and QDs are discussed.•This review covers analysis ...in both aqueous and organic eluents.•Challenges are addressed and future prospects outlined.
This review presents an overview of size-exclusion chromatographic separation and characterization of noble metal nanoparticles (NPs) and quantum dots (QDs) over the past 25 years. The properties of NPs and QDs that originate from quantum and surface effects are size dependent; to investigate these properties, a separation technique such as size-exclusion chromatography (SEC) is often needed to obtain narrow-distribution NP populations that are also separated from the unreacted starting materials. Information on the size distributions and optical properties of NPs has been obtained by coupling SEC to detection methods such as ultraviolet–visible and/or fluorescence spectroscopy. Problems associated with the sorption of NPs and QDs onto various SEC stationary phases, using both aqueous and organic eluents, are also discussed.
Our previous studies showed that particular antibiotic resistance genes (ARGs) were enriched locally in sediments below fish farms in the Northern Baltic Sea, Finland, even when the selection ...pressure from antibiotics was negligible. We assumed that a constant influx of farmed fish feces could be the plausible source of the ARGs enriched in the farm sediments. In the present study, we analyzed the composition of the antibiotic resistome from the intestinal contents of 20 fish from the Baltic Sea farms. We used a high-throughput method, WaferGen qPCR array with 364 primer sets to detect and quantify ARGs, mobile genetic elements (MGE), and the 16S rRNA gene. Despite a considerably wide selection of qPCR primer sets, only 28 genes were detected in the intestinal contents. The detected genes were ARGs encoding resistance to sulfonamide (
), trimethoprim (
), tetracycline
, aminoglycoside (
), chloramphenicol (
), and efflux-pumps resistance genes (
). The detected genes also included class 1 integron-associated genes (
Δ
) and transposases (
). Importantly, most of the detected genes were the same genes enriched in the farm sediments. This preliminary study suggests that feces from farmed fish contribute to the ARG enrichment in farm sediments despite the lack of contemporaneous antibiotic treatments at the farms. We observed that the intestinal contents of individual farmed fish had their own resistome compositions. Our result also showed that the total relative abundances of transposases and
genes were significantly correlated (
= 0.001,
= 0.71). In addition, we analyzed the mucosal skin and gill filament resistomes of the farmed fish but only one multidrug-efflux resistance gene (
) was detected. To our knowledge, this is the first study reporting the resistome of farmed fish using a culture-independent method. Determining the possible sources of ARGs, especially mobilized ARGs, is essential for controlling the occurrence and spread of ARGs at fish farming facilities and for lowering the risk of ARG spread from the farms to surrounding environments.
•MTG & trypsin modification enhanced instant exposure of OG’s hydrophobic side chain.•Procyanidin B2 acted as an acyl acceptor in the deamidation of MTG treated OG.•Both procyanidin B2 and enzymatic ...treated OG showed anti-aggregation properties.•SEC & AF4 are useful size information tools for large protein–phenolic aggregates.
The effect of enzyme treatment on protein–tannin interactions was investigated using up-to-date analytical approaches for improving their physical properties. The formation of ligands between procyanidin B2 and native oat globulin (OG) was observed to be affected by the ratio of procyanidin B2 to OG and the availability of tryptophan. For the transglutaminase-treated OG, the results obtained from circular dichroism (CD) and size exclusion chromatography (SEC) revealed that procyanidin B2 acted as an acyl acceptor in the process of OG deamidation. Procyanidin B2 also inhibited the non-covalent protein–protein interactions occurring between the aromatic side-chains or sedimentation of tryptophan aggregates. For trypsin-treated OG, procyanidin B2 interacted with phenylalanine and the tryptophan side-chain of OG. The inhibition of procyanidin B2 towards protein–protein aggregation was proved by the observation of CD, SEC and asymmetric flow field-flow fractionation.
Many microbial communities are characterized by high genetic diversity. 16S ribosomal RNA sequencing can determine community members, and metagenomics can determine the functional diversity, but ...resolving the functional role of individual cells in high throughput remains an unsolved challenge. Here, we describe epicPCR (Emulsion, Paired Isolation and Concatenation PCR), a new technique that links functional genes and phylogenetic markers in uncultured single cells, providing a throughput of hundreds of thousands of cells with costs comparable to one genomic library preparation. We demonstrate the utility of our technique in a natural environment by profiling a sulfate-reducing community in a freshwater lake, revealing both known sulfate reducers and discovering new putative sulfate reducers. Our method is adaptable to any conserved genetic trait and translates genetic associations from diverse microbial samples into a sequencing library that answers targeted ecological questions. Potential applications include identifying functional community members, tracing horizontal gene transfer networks and mapping ecological interactions between microbial cells.
The asymmetric flow field-flow fractionation (AF4) method was developed for cationic cellulose derivatives. AF4 is the method of choice especially for high-molar mass samples, which are challenging ...to characterize with conventional chromatographic techniques such as size-exclusion chromatography (SEC). The cationic charge of macromolecules also complicates the size-based separations where no interaction between the analytes and the column stationary phase (SEC) or membrane (AF4) should occur. However, many column matrices and membranes carry negative charge and thus preventing interactions between cationic analytes and negatively charged separation support should be taken into consideration when doing method development. In this study, two eluent compositions, neutral and acidic, were tested for AF4 separation of cationic hydroxyethyl celluloses with varying charge densities. The eluent composition with a pH below the isoelectric point of regenerated cellulose membrane, which was used in this AF4 study, enabled the size-based separation with close to 100% analysis recovery. Macromolecular parameters (molar mass and radius of gyration) and conformation were investigated by coupling a multi-angle light scattering detector and differential refractometer to the AF4 system.
Low temperature pre-treatments increase the char yield during cellulose carbonization. Although this effect is mostly understood on a kinetic basis, the formed chemical structures leave room for ...scrutiny. In ongoing ambitions to enhance the char yield of bioderived precursors, the thereby occurring chemistry was reinvestigated. A set of isothermal heating protocols ranging from 150° to 250°C was applied to man-made Ioncell® cellulose fibers and to Avicel® PH-101 microcrystalline cellulose. The prepared cellulosic samples were examined by solution state NMR using a tetra-n-butyl phosphonium acetate (P4444OAc): DMSO-d6 (1:4 wt%) electrolyte as dissolving medium. Complementary, IR spectroscopy, size-exclusion chromatography (SEC) and thermogravimetric analysis coupled with mass spectroscopy (TGA-MS) measurements were performed. The NMR spectra evidenced the formation of levoglucosan end capped moieties as being the first and major occurring reaction during low temperature pre-treatments. In contrast to other mechanistic proposals, no signs for carbonyl or alkene functionalities were discernible in the cellulosic material soluble in the NMR electrolyte, even after treatment at 250 °C for several hours. Thermal cross-linking was observed in SEC already at temperatures not known to significantly influence the overall char yield. In the solution state analytics only partial solubility was observed, owing to the formation of a reluctant fraction previously described as “thermostable condensed phase”. This resulted in analytical blind spots and led to discrepancies between NMR results and FTIR spectra in which carbonyl and alkene vibrations were clearly discernible. Those discrepancies might also imply the co-existence of different fractions in the early stages of cellulose pyrolysis.
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•Application of solution state analytics to heat pretreated cellulose fibers.•Evidence for levoglucosan end capped cellulose structures by solution state NMR.•Identification of crosslinked polysaccharide fractions by GPC.•Formation of a thermostable condensed phase as important carbonization intermediate.
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•Two isolated dextrans behaved differently in fava bean protein isolate (FPI).•Dextran (DX) from Weissella cibaria Sj 1b showed a high gel-strengthening ability.•Maillard reaction ...showed different effects on rheology depending on the system.•Intermolecular FPI-DX interactions play a major role in stabilizing protein network.
The aim of this study was to study the interactions between dextran and fava bean protein. Two dextrans produced by Leuconostoc pseudomesenteroides DSM 20193 and Weissella cibaria Sj 1b were purified and mixed with fava bean protein isolate (FPI) in water or in different buffers. The two isolated dextrans presented a typical dextran structure, mainly α-(1 → 6) linkages (above 95%) and few α-(1 → 3) branches, but they differed in molar mass and conformation. Dry-heating incubation of FPI and dextran mixture facilitated the conjugation of dextran to FPI through the Maillard reaction. Both mixed and conjugated systems were further heat-treated, and different influences of the formed covalent bonds on rheological properties were observed. The W. cibaria Sj 1b dextran had a much higher gel-strengthening ability than the Ln. pseudomesenteroides DSM 20193 dextran. The intermolecular FPI-dextran interactions played an important role in stabilizing the mixed systems at different pH.
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