Covalent association of LTBP-1 (latent TGF-beta binding protein-1) to latent TGF-beta is mediated by the third eight-cysteine (also referred to as TB) module of LTBP-1, a domain designated as CR3. Spodoptera frugiperda (Sf9) cells have proved a suitable cell system in which to study this association and to produce recombinant CR3, and we show here that another lepidopteran cell line, Trichoplusia niTN-5B1-4 (High-Five) cells, allows the recovery of large amounts of functional recombinant CR3. CR3 contains an N-glycosylation site, which is conserved in all forms of LTBP known to date. When we examined the status of this N-glycosylation using MALDI-TOF mass spectrometry and enzymatic analysis, we found that CR3 is one of the rare recombinant peptides modified with complex glycans in insect cells. Sf9 cells mainly processed the fucosylated paucomannosidic structure (GlcNAc)(2)(Mannose)(3)Fucose, although hybrid and complex N-glycosylations were also detected. In High-Five cells, the peptide was found to be modified with a wide variety of hybrid and complex sugars in addition to paucomanosidic oligosaccharides. Most glycans had one or two fucose residues bound through alpha1,3 and alpha1,6 linkages to the innermost GlcNAc. On the basis of these results and on the structure of an eight-cysteine domain from fibrillin-1, we present a model of glycosylated CR3 and discuss the role of glycosylation in eight-cysteine domain protein-protein interactions.
This study assessed changes in the phytoplankton community related to macronutrient and bioactive trace metal distribution in surface waters of the Atlantic sector of the Southern Ocean, between Cape Town and Antarctica along the GEOTRACES GIPY_05 (mainly along prime meridian) transect in summer 2014-2015. Several general community structure features were reaffirmed, such as the restriction of cyanobacteria to the northern Subtropical Zone, while haptophytes, such as Phaeocystis, along with diatoms, dominate the community north of the Polar Front, and diatoms clearly dominate south of the Polar Front. These community structure changes were often linked with macro-and micro-nutrient composition changes. For example, the concentration of diatoms increased southwards with the availability of silica whereas the cyanobacterial contribution in the northern Subtropical Zone appeared to be linked with labile cobalt depletion. However, these links were not consistent along the entire transect, i.e., no individual nutrient, such as silica or iron, was linked to community composition changes across all water masses. Each station showed a rather unique combination of nutrient and community compositions. Our findings also indicated impacts on the phytoplankton community through trace metal distributions that could be related to a deep mixing event at ∼54 • S and to ice melt at ∼65 and 68 • S. The timing of sampling after such trace metal fluxes proved to be an important consideration, particularly where iron appeared to be preferentially depleted to near-limiting concentrations, possibly driving utilization of other metals. The study highlights the importance of considering a suite of trace metals when assessing controls of phytoplankton variability in the open ocean and emphasizes the need for higher resolution trace metal sampling and multi-element incubation studies to further study the complex relationships between phytoplankton and nutrients.
Integrating renewable energy into public space is becoming more common as a climate change solution. However, this approach is often guided by the environmental pillar of sustainability, with less focus on the economic and social pillars. The purpose of this paper is to examine this issue in the speculative renewable energy propositions for Freshkills Park in New York City submitted for the 2012 Land Art Generator Initiative (LAGI) competition. This paper first proposes an optimal electricity distribution (OED) framework in and around public spaces based on relevant ecology and energy theory (Odum's fourth and fifth law of thermodynamics). This framework addresses social engagement related to public interaction, and economic engagement related to the estimated quantity of electricity produced, in conjunction with environmental engagement related to the embodied energy required to construct the renewable energy infrastructure. Next, the study uses the OED framework to analyse the top twenty-five projects submitted for the LAGI 2012 competition. The findings reveal an electricity distribution imbalance and suggest a lack of in-depth understanding about sustainable electricity distribution within public space design. The paper concludes with suggestions for future research.
Fusobacterium necrophorum is a strict anaerobic organism responsible for a number of clinical syndromes known as necrobacillosis. Although meningeal infections with anaerobes are rare, delayed diagnosis and treatment can be potentially fatal. We report a unique case of Fusobacterium meningitis, mastoiditis, and sepsis in a previously healthy adolescent. Diagnosis and management of this condition are discussed in the context of a literature review.
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