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Marine pelagic mucilages (e.g., marine snow) have been reported to a greater extent than their lacustrine counterparts. A pelagic mucilage primarily comprised of chitin secretions from the invasive centric diatom, Lindavia intermedia, has been reported since the early 2000s, primarily from large, oligotrophic pre-alpine lakes of the South Island of New Zealand. To better understand the factors related to mucilage abundance, we monitored its abundance as well as factors potentially related to mucilage production over time in four mucilage-afflicted lakes. Temporal mucilage dynamics were episodic, with peaks in abundance occurring during any season, but most often during summer and autumn. Chitin was confirmed to be an important component of the mucilage, but the chitin content varied between 1 and 12% of the mucilage dry mass in the lakes. An RT-qPCR assay for chitin synthase gene overexpression in L. intermedia showed that overexpression occurred in summer and autumn, often when peaks in mucilage abundance also occurred. A correlation between mucilage and phytoplankton abundance was only observed in one of the lakes. Both dissolved reactive and total phosphorus concentrations were often below analytical detection limits in these lakes. Nitrate concentrations were also low and showed negative correlations with mucilage abundance. This suggests either that the secrection of chitinous mucilage by L. intermedia significantly depleted the available N in the water column or that mucilage facilitated N uptake by L. intermedia and/or other microorganisms associated with the mucilage. Pelagic mucilage in New Zealand lakes shares many characteristics of other conspicuous mucilage phenomena, including lake snow and marine snow. While our correlational analyses revealed some relationships and associations with mucilage abundance, the strengths of these were quite variable, indicating that as yet unstudied mucilage loss processes in these lakes (e.g., sedimentation, disintegration, decomposition, assimilation) likely also play important roles in regulating mucilage abundance. [ABSTRACT FROM AUTHOR]

Objective: Packable composites are a comparatively recent addition to the dentist's armamentarium, Solitaire-2 being an example of this type of material. This paper reports the performance of 100 restorations formed in Solitaire-2 in conjunction with the Gluma Solid Bond system and Gluma One Bond bonding systems, in Class I and II cavity restorations in permanent teeth, placed in the practices of five members of the Product Research and Evaluation by Practitioners (PREP) Panel, a group of United Kingdom-based dental practitioners who are prepared to undertake research projects in their practices. Method and materials: Five members of the PREP Panel were each requested to place 20 Solitaire-2 restorations. These restorations were reviewed at 1 year by a trained and calibrated evaluator, and the PREP panel member who had placed the restorations. Results: A total of 88 restorations (33 Class I, 55 Class II in 49 patients (mean age 43 years) were reviewed at 1 year. One Class II restoration (a large mesio-occlusodistal restoration) had been replaced at 10 months after a fracture was detected across the distal box. The remaining 87 (99%) of the restorations were intact with no secondary caries detected. Conclusion: Ninety-nine percent of the Solitaire-2 restorations, placed in general dental practice conditions in conjunction with the Gluma Solid Bond system and Gluma One Bond bonding systems, were found to be performing satisfactorily at 1 year. [ABSTRACT FROM AUTHOR]

Objective: The purpose of this study was to assess Pertac II restorations placed in general dental practice. Method and materials: A total of 86 restorations (14 Class 1, 10 Class II, 17 Class III, 5 Class IV, and 40 Class V) using a hybrid composite material (Pertac II) placed in 56 patients (mean age, 39 years) in 5 dental practices in the United Kingdom were assessed after 1 year by a trained evaluator and the dental practitioner who had placed the restoration. All the restorations were assessed for anatomic form, marginal adaptation, surface roughness, sensitivity or discomfort, gingival condition, and the presence or absence of secondary caries. Results: Thirty-five (43%) restorations were placed in load-bearing situations, 9% of which were assessed as being in "heavy" occlusion. Five (6%) of the restorations were lost (all Class V restorations), 4 from premolars and 1 from a molar. Four were in wedge-shaped cavities and 1 small restoration was actually dislodged during examination. The remaining 81 (94%) restorations were found to be intact, with no secondary caries detected. A high percentage of optimal scores were recorded for both anatomic form and surface roughness. Conclusion: It is concluded that the evaluation demonstrated satisfactory clinical performance of Pertac II restorations after 1 year of clinical service in the wide variety of clinical situations seen in general dental practice. However, the potential for failure of hybrid composite materials in Class V situations appears high. It may be argued that clinicians who wish to use a resin-based material in a Class V situation should use a compomer or microfilled composite. [ABSTRACT FROM AUTHOR]

The future of human mesenchymal stem cells (hMSCs) as a successful cell therapy relies on bioprocessing strategies to improve the scalability of these cells without compromising their therapeutic ability. The culture-expansion of hMSCs can be enhanced by supplementation with growth factors, particularly fibroblast growth factor 2 (FGF2). The biological activity of FGF2 is controlled through interactions with heparan sulfate (HS) that facilitates ligand-receptor complex formation. We previously reported on an FGF2-interacting HS variant (termed HS2) isolated from embryonic tissue by anionic exchange chromatography that increased the proliferation and potency of hMSCs. Here, we detail the isolation of an FGF2 affinity-purified HS variant (HS8) using a scalable platform technology previously employed to generate HS variants with increased affinity for BMP-2 or VEGF165. This process used a peptide sequence derived from the heparin-binding domain of FGF2 as a substrate to affinity-isolate HS8 from a commercially available source of porcine mucosal HS. Our data show that HS8 binds to FGF2 with higher affinity than to FGF1, FGF7, BMP2, PDGF-BB, or VEGF165. Also, HS8 protects FGF2 from thermal destabilization and increases FGF signaling and hMSC proliferation through FGF receptor 1. Long-term supplementation of cultures with HS8 increased both hMSC numbers and their colony-forming efficiency without adversely affecting the expression of hMSC-related cell surface antigens. This strategy further exemplifies the utility of affinity-purifying HS variants against particular ligands important to the stem cell microenvironment and advocates for their addition as adjuvants for the culture-expansion of hMSCs destined for cellular therapy. J. Cell. Physiol. 232: 566-575, 2017. © 2016 Wiley Periodicals, Inc. [ABSTRACT FROM AUTHOR]

The depolymerisation of porcine mucosal heparan sulfate under the action of heparin lyases and analysis by size-exclusion chromatography (SEC) is described. Heparan sulfate treated to enzymic bond scission producing a Δ 4,5 double-bond and quantified by SEC with ultraviolet-visible (UV) spectroscopic detection (230 nm) indicated that the majority of the biopolymer (>85%) was reduced to disaccharides (degree of polymerisation (DP) = 2). However, analysis of the SEC eluant using refractive index (RI), which reflects the mass contribution of the oligosaccharides rather than the molar response of a UV chromophore, indicated that a considerable proportion of the digested HS, up to 43%, was present with DP >2. This was supported by a mass balance analysis. These results contradict the accepted literature where “complete digestion” is routinely reported. Herein we report on the composition and methodology utilised to ascertain the extent of depolymerization and disaccharide composition of this important biopolymer. [ABSTRACT FROM AUTHOR]

Water-soluble, non-starch polysaccharides from plants are used commercially in a wide range of food and non-food applications. The increasing range of applications for natural polysaccharides means that there is growing demand for plant-derived polysaccharides with different functionalities. The geographical isolation of New Zealand and its unique flora presents opportunities to discover new polysaccharides with novel properties for a range of applications. This review brings together data published since the year 2000 on the composition and structure of exudate gums, mucilages, and storage polysaccharides extracted from New Zealand endemic land plants. The structures and properties of these polysaccharides are compared with the structures of similar polysaccharides from other plants. The current commercial use of these polysaccharides is reviewed and their potential for further exploitation discussed. [ABSTRACT FROM AUTHOR]

Dietary fiber provides growth substrates for bacterial species that belong to the colonic microbiota of humans. The microbiota degrades and ferments substrates, producing characteristic short-chain fatty acid profiles. Dietary fiber contains plant cell wall-associated polysaccharides (hemicelluloses and pectins) that are chemically diverse in composition and structure. Thus, depending on plant sources, dietary fiber daily presents the microbiota with mixtures of plant polysaccharides of various types and complexity. We studied the extent and preferential order in which mixtures of plant polysaccharides (arabinoxylan, xyloglucan, β-glucan, and pectin) were utilized by a coculture of five bacterial species (Bacteroides ovatus, Bifidobacterium longum subspecies longum, Megasphaera elsdenii, Ruminococcus gnavus, and Veillonella parvula). These species are members of the human gut microbiota and have the biochemical capacity, collectively, to degrade and ferment the polysaccharides and produce short-chain fatty acids (SCFAs). B. ovatus utilized glycans in the order β-glucan, pectin, xyloglucan, and arabinoxylan, whereas B. longum subsp. longum utilization was in the order arabinoxylan, arabinan, pectin, and β-glucan. Propionate, as a proportion of total SCFAs, was augmented when polysaccharide mixtures contained galactan, resulting in greater succinate production by B. ovatus and conversion of succinate to propionate by V. parvula. Overall, we derived a synthetic ecological community that carries out SCFA production by the common pathways used by bacterial species for this purpose. Systems like this might be used to predict changes to the emergent properties of the gut ecosystem when diet is altered, with the aim of beneficially affecting human physiology. [ABSTRACT FROM AUTHOR]

Whole-transcriptome analysis was used to investigate the molecular interplay between three bacterial species that are members of the human gut microbiota. Bacteroides ovatus , Subdoligranulum variabile , and Hungatella hathewayi formed associations in cocultures fed barley β-glucan, a constituent of dietary fiber. B. ovatus depolymerized β-glucan and released, but did not utilize, 3- O -β-cellobiosyl-d-glucose (DP3) and 3- O -β-cellotriosyl-d-glucose (DP4). These oligosaccharides provided growth substrates for S. variabile and H. hathewayi with a preference for DP4 in the case of the latter species. There was increased transcription of a B. ovatus mixed-linkage-β-glucan utilization locus, as well as carbohydrate transporters in S. variabile and H. hathewayi when in batch coculture. Increased transcription of the β-glucan utilization locus did not occur in continuous culture. Evidence for interactions relating to provision of cobalamin, alterations to signaling, and modulation of the "stringent response" (an adaptation to nutrient deprivation) were detected. Overall, we established a bacterial consortium based on barley β-glucan in vitro , which can be used to investigate aspects of the functional blueprint of the human gut microbiota. IMPORTANCE The microbial community, mostly composed of bacterial species, residing in the human gut degrades and ferments polysaccharides derived from plants (dietary fiber) that would not otherwise be digested. In this way, the collective metabolic actions of community members extract additional energy from the human diet. While the variety of bacteria present in the microbial community is well known, the formation of bacterial consortia, and the consequent interactions that result in the digestion of dietary polysaccharides, has not been studied extensively. The importance of our work was the establishment, under laboratory conditions, of a consortium of gut bacteria that formed around a dietary constituent commonly present in cereals. This enabled the metabolic interplay between the bacterial species to be studied. This kind of knowledge is required to construct an interactive, metabolic blueprint of the microbial community that inhabits the human gut., (Copyright © 2020 American Society for Microbiology.)

Dietary fiber provides growth substrates for bacterial species that belong to the colonic microbiota of humans. The microbiota degrades and ferments substrates, producing characteristic short-chain fatty acid profiles. Dietary fiber contains plant cell wall-associated polysaccharides (hemicelluloses and pectins) that are chemically diverse in composition and structure. Thus, depending on plant sources, dietary fiber daily presents the microbiota with mixtures of plant polysaccharides of various types and complexity. We studied the extent and preferential order in which mixtures of plant polysaccharides (arabinoxylan, xyloglucan, β-glucan, and pectin) were utilized by a coculture of five bacterial species ( Bacteroides ovatus , Bifidobacterium longum subspecies longum , Megasphaera elsdenii , Ruminococcus gnavus , and Veillonella parvula ). These species are members of the human gut microbiota and have the biochemical capacity, collectively, to degrade and ferment the polysaccharides and produce short-chain fatty acids (SCFAs). B. ovatus utilized glycans in the order β-glucan, pectin, xyloglucan, and arabinoxylan, whereas B. longum subsp. longum utilization was in the order arabinoxylan, arabinan, pectin, and β-glucan. Propionate, as a proportion of total SCFAs, was augmented when polysaccharide mixtures contained galactan, resulting in greater succinate production by B. ovatus and conversion of succinate to propionate by V. parvula Overall, we derived a synthetic ecological community that carries out SCFA production by the common pathways used by bacterial species for this purpose. Systems like this might be used to predict changes to the emergent properties of the gut ecosystem when diet is altered, with the aim of beneficially affecting human physiology. IMPORTANCE This study addresses the question as to how bacterial species, characteristic of the human gut microbiota, collectively utilize mixtures of plant polysaccharides such as are found in dietary fiber. Five bacterial species with the capacity to degrade polymers and/or produce acidic fermentation products detectable in human feces were used in the experiments. The bacteria showed preferential use of certain polysaccharides over others for growth, and this influenced their fermentation output qualitatively. These kinds of studies are essential in developing concepts of how the gut microbial community shares habitat resources, directly and indirectly, when presented with mixtures of polysaccharides that are found in human diets. The concepts are required in planning dietary interventions that might correct imbalances in the functioning of the human microbiota so as to support measures to reduce metabolic conditions such as obesity., (Copyright © 2020 American Society for Microbiology.)

A knowledge of the mobilities of the polysaccharides or parts of polysaccharides in a cell-wall preparation provides information about possible molecular interactions among the polysaccharides in the cell wall and the relative locations of polysaccharides within the cell wall. A number of solid-state 13 C NMR techniques have been developed that can be used to investigate different types of polysaccharide mobilities: rigid, semirigid, mobile, and highly mobile. In this chapter techniques are described for obtaining spectra from primary cell-wall preparations using CP/MAS, proton-rotating frame, proton spin-spin, spin-echo relaxation spectra and single-pulse excitation. We also describe how proton spin relaxation editing can be used to obtain subspectra for cell-wall polysaccharides of different mobilities, and how 2D and 3D solid-state NMR experiments have recently been applied to plant cell walls.

Pectic polysaccharides from New Zealand (NZ) spinach ( Tetragonia tetragonioides ) and karaka berries ( Corynocarpus laevigatus ) were extracted and analyzed. NZ spinach polysaccharides comprised mostly homogalacturonan (64.4%) and rhamnogalacturonan I (5.8%), with side chains of arabinan (8.1%), galactan (2.2%), and type II arabinogalactan (7.1%); karaka berry polysaccharides comprised homogalacturonan (21.8%) and rhamnogalacturonan I (10.0%), with greater proportions of side chains (arabinan, 15.6%; galactan, 23.8%; and type II arabinogalactan, 19.3%). Screening of gut commensal Bacteroides showed that six were able to grow on the NZ spinach extract, while five were able to grow on the karaka berry extract. Analysis of the polysaccharides remaining after fermentation, by size-exclusion chromatography and constituent sugar analysis, showed that the Bacteroides species that grew on these two substrates showed preferences for the different pectic polysaccharide types. Our data suggest that, to completely degrade and utilize the complex pectin structures found in plants, members of Bacteroides and other bowel bacteria work as metabolic consortia.

Polysaccharides from feijoa fruit were extracted and analyzed; the composition of these polysaccharides conforms to those typically found in the primary cell walls of eudicotyledons. The two major polysaccharide extracts consisted of mainly pectic polysaccharides and hemicellulosic polysaccharides [xyloglucan (77%) and arabinoxylan (16%)]. A collection of commensal Bacteroides species was screened for growth in culture using these polysaccharide preparations and placed into five categories based on their preference for each substrate. Most of the species tested could utilize the pectic polysaccharides, but growth on the hemicellulose was more limited. Constituent sugar and glycosyl linkage analysis showed that species that grew on the hemicellulose fraction showed differences in their preference for the two polysaccharides in this preparation. Our data demonstrate that the members of the genus Bacteroides show differential hydrolysis of pectic polysaccharides, xyloglucan, and arabinoxylan, which might influence the structure and metabolic activities of the microbiota in the human gut.

A novel anaerobic pectinolytic bacterium (strain 14 T ) was isolated from human faeces. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain 14 T belonged to the family Ruminococcaceae, but was located separately from known clostridial clusters within the taxon. The closest cultured relative of strain 14 T was Acetivibrio cellulolyticus (89.7 % sequence similarity). Strain 14 T shared ~99 % sequence similarity with cloned 16S rRNA gene sequences from uncultured bacteria derived from the human gut. Cells were Gram-stain-positive, non-motile cocci approximately 0.6 µm in diameter. Strain 14 T fermented pectins from citrus peel, apple, and kiwifruit as well as carbohydrates that are constituents of pectins and hemicellulose, such as galacturonic acid, xylose, and arabinose. TEM images of strain 14 T , cultured in association with plant tissues, suggested extracellular fibrolytic activity associated with the bacterial cells, forming zones of degradation in the pectin-rich regions of middle lamella. Phylogenetic and phenotypic analysis supported the differentiation of strain 14 T as a novel genus in the family Ruminococcaceae. The name Monoglobus pectinilyticus gen. nov., sp. nov. is proposed; the type strain is 14 T (JCM 31914 T =DSM 104782 T ).