Identification of blottin: a novel gastric trefoil factor family-2 binding protein

The trefoil factor family (TFF) peptides are important in gastro-intestinal mucosal protection and repair. Their mechanism of action remains unclear and receptors are sought. We aimed to identify and characterise proteins binding to TFF2. A fusion protein of mouse TFF2 with alkaline phosphatase was generated and used to probe 2-D protein blots of mouse stomach. The resulting spots were analysed by MS. The protein identified was characterised by bioinformatics, rapid amplification of cDNA ends, in situ hybridisation (ISH) and immunohistochemistry (IHC). Functional assays were performed in gastrointestinal cell lines. A single major murine protein was identified and named blottin. It was previously unknown as a translated product. Blottin is also present in rat and human; the latter gene is also known as GDDR. The predicted full-length proteins are 184 amino acids long (20 kDa), reducing to 164 amino acids (18 kDa) after signal peptide cleavage. ISH of gastrointestinal tissues shows abundant blottin mRNA in gastric surface and foveolar epithelium. IHC shows cytoplasmic staining for blottin protein, and by immunoelectron microscopy in mucus granules and Golgi stacks. Previous work showed that blottin is down-regulated in gastric cancers. Blottin contains a BRICHOS domain, and has 56% similarity with gastrokine-1. Cultured HT-29 cells express blottin and show increased DNA synthesis with antiblottin antibody; however, this effect is reversed by the immunising peptide. We have identified and characterised a TFF2-binding protein produced by gastric epithelium. Blottin may play a role in gastrointestinal mucosal protection and modulate gut epithelial cell proliferation.     

Immobilization of enzymes based on hydrophobic interaction. III. Adsorbent substituent density and its impact on the immobilization of β-amylase

Hexyl-groups have been introduced into crosslinked Sepharose 6B, yielding gels with degrees of substitution which range from 0.02 to 0.70 mol hexyl-side chain per mole galactose residue. The gels were exposed to β-amylase in solution, and the resulting adsorbates indicated a monotonic increase in adsorption capacity with an increasing hexyl-content. Adsorbate activity, by contrast, displayed a maximum for a carrier gel with a hexyl–galactose ratio of 0.51. Adsorbates based on gels with different hexyl-content were used in column reactors for continuous maltose production from a soluble starch substrate.     

Anatomical and molecular characterization of Lactarius aff. omphaliformis, Russula alnijorullensis and Cortinarius tucumanensis ectomycorrhizae on Alnus acuminata

Ectomycorrhizae (ECM) of Lactarius aff. omphaliformis Romagn., Russula alnijorullensis (Sing.) Sing. and Cortinarius tucumanensis Mos. on Andean alder (Alnus acuminata Kunth) were characterized and identified. The identification of the fungal symbionts was achieved by morpho-anatomical observations of mycorrhizae and by comparison of ITS-RFLP patterns obtained from ECM and fruitbodies. L. aff omphaliformis ECM differed in some morphological details such as ramification and mantle type from ECM of the same species on A. glutinosa. L. aff omphaliformis ECM show an orange to ochre mantle containing latex cells, which stain with sulpho-vanillin, emanating hyphae without clamps. R. alnijorullensis ECM represent a typical Russula-type-ECM, light yellow to pinkish, the outer mantle being composed of triangular latex-filled cells staining with sulpho-vanillin, emanating hyphae without clamps. C. tucumanensis ECM exhibit a white (silvery) to yellowish brown mantle covered with soil particles, emanating hyphae with clamps.     

Testing the intermediate disturbance hypothesis: when will there be two peaks of diversity?

Succession after disturbances generates a mosaic of patches in different successional stages. The intermediate disturbance hypothesis predicts that intermediate disturbances lead to the highest diversity of these stages on a regional scale resulting in a hump‐shaped diversity–disturbance curve. We tested this prediction using field data of forest succession and hypothetical succession scenarios in combination with analytical and simulation models. According to our study the main factors shaping the diversity–disturbance curve and the position of the diversity maximum were the transition times between the successional stages, the transition type, neighbourhood effects and the choice of diversity measure. Although many scenarios confirmed the intermediate disturbance hypothesis we found that deviations in the form of two diversity maximums were possible. Such bimodal diversity–disturbance curves occurred when early and late successional stages were separated by one or more long‐lived (compared to the early stages) intermediate successional stages. Although the field data which met these conditions among all those tested were rare (one of six), the consequences of detecting two peaks are fundamental. The impact of disturbances on biodiversity can be complex and deviate from a hump‐shaped curve.     

Mapping changes to vegetation pattern in a restoring wetland: Finding pattern metrics that are consistent across spatial scale and time

Tidal salt marshes in the San Francisco Estuary region display heterogeneous vegetation patterns that influence wetland function and provide adequate habitat for native or endangered wildlife. In addition to analyzing the extent of vegetation, monitoring the dynamics of vegetation pattern within restoring wetlands can offer valuable information about the restoration process. Pattern metrics, derived from classified remotely sensed imagery, have been used to measure composition and configuration of patches and landscapes, but they can be unpredictable across scales, and inconsistent across time. We sought to identify pattern metrics that are consistent across spatial scale and time – and thus robust measures of vegetation and habitat configuration – for a restored tidal marsh in the San Francisco Bay, CA, USA. We used high-resolution (20 cm) remotely sensed color infrared imagery to map vegetation pattern over 2 years, and performed a multi-scale analysis of derived vegetation pattern metrics. We looked at the influence on metrics of changes in grain size through resampling and changes in minimum mapping unit (MMU) through smoothing. We examined composition, complexity, connectivity and heterogeneity metrics, focusing on perennial pickleweed (Sarcocornia pacifica), a dominant marsh plant. At our site, pickleweed patches grew larger, more irregularly shaped, and closely spaced over time, while the overall landscape became more diverse. Of the two scale factors examined, grain size was more consistent than MMU in terms of identifying relative change in composition and configuration of wetland marsh vegetation over time. Most metrics exhibited unstable behavior with larger MMUs. With small MMUs, most metrics were consistent across grain sizes, from fine (e.g. 0.16 m²) to relatively large (e.g. 16 m²) pixel sizes. Scale relationships were more variable at the landcover class level than at the landscape level (across all classes). This information may be useful to applied restoration practitioners, and adds to our general understanding of vegetation change in a restoring marsh.     

Adsorption of HSA, IgG and laminin-1 on model hydroxyapatite surfaces--effects of surface characteristics

Ellipsometry and mechanically assisted sodium dodecyl sulphate elution was utilized to study the adsorption of human serum albumin (HSA), human immunoglobulin G (IgG), and laminin-1, as well as competitive adsorption from a mixture of these proteins on spin-coated and sintered hydroxyapatite (HA) surfaces, respectively. The HA surfaces were characterized with respect to wettability and roughness by means of water contact angles and atomic force microscopy, respectively. Both surface types were hydrophilic, and the average roughness (Sa) and surface enlargement (Sdr) were lower for the sintered compared to the spin-coated HA surfaces. The adsorbed amounts on the sintered HA increased as follows: HSA < laminin-1 < IgG < the protein mixture. For the competitive adsorption experiments, the adsorbed fractions increased accordingly: HSA < laminin-1 < IgG on both types of HA substratum. However, a higher relative amount of HSA and laminin-1 and a lower relative amount of IgG was found on the spin-coated surfaces compared to the sintered surfaces. The effects observed could be ascribed to differences in surface roughness and chemical composition between the two types of HA substratum, and could have an influence on selection of future implant surface coatings.     

Transferability of Material Composition Indicators for Residential Buildings: A Conceptual Approach Based on a German‐Japanese Comparison

Most anthropogenic material stocks and flows are associated with the building sector. Several recent studies have developed material composition indicators (MCIs) suitable for calculating material stocks and flows of the building sector using bottom‐up approaches, which hold great potential to provide information to support resource efficiency policies. A major limitation is the lack of country‐specific MCIs. This study aims to introduce a concept for a better transferability of MCI across different contexts by proposing requirements for defining MCIs and to discuss options and limits of the transferability. We take existing MCIs for residential buildings in Germany and Japan as case studies and make them comparable by applying harmonization methods. Based on that, similarities and differences are systematically identified and discussed, considering their socioeconomic, cultural, technical, and environmental factors. Our results indicate significant limitations to the transferability of MCIs for detached houses, while bigger apartment complexes show greater homogeneity despite the very different environments in which they are constructed. This indicates that while it is possible to assume foreign MCIs as plausible for large constructions, local coefficients need to be estimated for smaller single‐family homes.     

Germline variants in oculocutaneous albinism genes and predisposition to familial cutaneous melanoma

Approximately 1%–2% of cutaneous melanoma (CM) is classified as strongly familial. We sought to investigate unexplained CM predisposition in families negative for the known susceptibility genes using next‐generation sequencing of affected individuals. Segregation of germline variants of interest within families was assessed by Sanger sequencing. Several heterozygous variants in oculocutaneous albinism (OCA) genes: TYR, OCA2, TYRP1 and SLC45A2, were present in our CM cohort. OCA is a group of autosomal recessive genetic disorders, resulting in pigmentation defects of the eyes, hair and skin. Missense variants classified as pathogenic for OCA were present in multiple families and some fully segregated with CM. The functionally compromised TYR p.T373K variant was present in three unrelated families. In OCA2, known pathogenic variants: p.V443I and p.N489D, were present in three families and one family, respectively. We identified a likely pathogenic SLC45A2 frameshift variant that fully segregated with CM in a family of four cases. Another four‐case family harboured cosegregating variants (p.A24T and p.R153C) of uncertain functional significance in TYRP1. We conclude that rare, heterozygous variants in OCA genes confer moderate risk for CM.     

Influences of blackberry margins on population dynamics of Drosophila suzukii and grape infestation in adjacent vineyards

Understanding the dynamics of pest insect populations in relation to the presence of non‐crop habitats and infestation levels of adjacent crops is essential to develop sustainable pest management strategies. The invasive pest species Drosophila suzukii (Diptera: Drosophilidae) is able to utilize a broad range of host plants. In viticulture, scientific risk assessment for D. suzukii has only recently started and studies assessing the effects of field margins containing wild host plants on D. suzukii population dynamics and on infestation risks in adjacent vineyards are lacking. Thus, in a one‐year field study, the role of different field margins on fly abundance and crop infestation in adjacent vineyards of Vitis vinifera, variety “Pinot Noir,” were investigated. Different monitoring methods were conducted to assess fly distribution, sex ratio and grape infestation in 14 vineyards adjacent to field margins containing either blackberry (BB) Rubus spp. or non‐host (NH) plants. Our results show that blackberries strongly enhanced D. suzukii abundance within field margin vegetation all year long, whereas fly abundance in vineyards adjacent to BB margins was just enhanced in some seasonal periods. Moreover, the influence of BB margins was limited by distance. However, high fly numbers in BB field margins did result in zero egg infestation of “Pinot Noir” berries. These results may have important implications for winegrowers to make efficient management decisions: regardless of high abundance of adult D. suzukii, only grape berry monitoring can assess the actual infestation risk and the potential need to take management action.     

De novo synthesis of NGF subunits in S-180 mouse sarcoma cell line

It is an accepted hypothesis that the nerve growth factor protein (NGF) plays an important role in the development of vertebrate sympathetic and sensory ganglia and has effects on some central neurons. The best known NGF species is that isolated from the mouse submaxillary gland, MSG-NGF. MSG-NGF can be isolated as a subunit containing protein, 7S-NGF, made up of three dissimilar subunits called alpha-, beta-, and gamma-NGF. Beta-NGF is the biologically active subunit and its synthesis in vivo and in vitro has been demonstrated. Less is known about the synthesis of the alpha- and gamma-NGF or the assembly of the subunits into the 7S complex. In order to develop a clonal model system for the study of NGF synthesis, processing and secretion, affinity chromatography techniques were applied to cell extracts of S180 mouse sarcoma, a cell line known to synthesize NGF. After incubating S180 cells in³⁵S-Methionine, cell extracts were exposed to antibody directed against alpha-NGF, gamma-NGF or beta-NGF covalently bound to Sepharose beads in order to elute and characterize the desired NGF subunits. Parallel experiments using immunoabsorbed [³⁵S]Methionine-beta-NGF were carried out in the presence or absence of excess NGF, in order to demonstrate the specificity of this procedure. Affinity chromatography with a substrate analogue to arginine ester bound to Sepharose beads was also used to isolate de novo synthesized gamma-NGF. We were able to show that the S180 line synthesized alpha-, beta-, and gamma-NGF indistiguishable from alpha-, beta-, and gamma-NGF isolated from mouse submaxillary gland in terms of antigenic and physicochemical properties, and biological and enzymatic activities. These results are consistent with the hypothesis that NGF is synthesized, assembled and secreted by a single cell type.Special Issue dedicated to Dr. E. M. Shooter and Dr. S. Varon.