Soil frost enhances stream dissolved organic carbon concentrations during episodic spring snow melt from boreal mires

Changes in winter time conditions at high‐latitude ecosystems could severely affect the carbon exchange processes. Using a 15 year stream record combined with winter field measurements and laboratory experiment, we studied the regulation of dissolved organic carbon (DOC) concentration in stream water draining boreal mire during snow melt. The most unanticipated finding was that cold soils with deep soil frost resulted in increased snow melt DOC concentrations in the stream runoff. Wintertime field measurements of DOC concentrations below the mire soil frost showed that this phenomenon could be explained by freeze‐out of DOC giving higher levels of DOC in the soil water below the ice as the soil frost developed downwards in the mire. Experimental freezing of water with a certain DOC concentration in the laboratory further corroborated the freeze‐out of DOC. In this experiment, as much as 96% of the DOC was excluded from the ice, whereas the freeze‐out in the mire was less effective (60%). The difference between the proportion of DOC retained in pure water relative to the proportion retained in peat water during freezing is probably due to trapped DOC in the solid peat soil matrix. A simple mass‐balance model showed that to explain the higher stream DOC concentrations during a winter with deep soil frost, approximately 0.5% of the mire area needed to be hydrologically connected to the stream discharge. In the stream records, we also found that the DOC concentrations during snow melt episodic runoff were negatively related to increasing intensity of the snow melt episodes (dilution by low DOC snow melt water) and higher previous export of DOC.     

Aquatic export of young dissolved and gaseous carbon from a pristine boreal fen: Implications for peat carbon stock stability

The stability of northern peatland's carbon (C) store under changing climate is of major concern for the global C cycle. The aquatic export of C from boreal peatlands is recognized as both a critical pathway for the remobilization of peat C stocks as well as a major component of the net ecosystem C balance (NECB). Here, we present a full year characterization of radiocarbon content (¹⁴C) of dissolved organic carbon (DOC), carbon dioxide (CO₂), and methane (CH₄) exported from a boreal peatland catchment coupled with ¹⁴C characterization of the catchment's peat profile of the same C species. The age of aquatic C in runoff varied little throughout the year and appeared to be sustained by recently fixed C from the atmosphere (<60 years), despite stream DOC, CO₂, and CH₄ primarily being sourced from deep peat horizons (2–4 m) near the mire's outlet. In fact, the ¹⁴C content of DOC, CO₂, and CH₄ across the entire peat profile was considerably enriched with postbomb C compared with the solid peat material. Overall, our results demonstrate little to no mobilization of ancient C stocks from this boreal peatland and a relatively large resilience of the source of aquatic C export to forecasted hydroclimatic changes.     

The Synaptonemal Complex Protein SCP3 Can Form Multistranded, Cross-striated Fibers In Vivo

The synaptonemal complex protein SCP3 is part of the lateral element of the synaptonemal complex, a meiosis-specific protein structure essential for synapsis of homologous chromosomes. We have investigated the fiber-forming properties of SCP3 to elucidate its role in the synaptonemal complex. By synthesis of SCP3 in cultured somatic cells, it has been shown that SCP3 can self-assemble into thick fibers and that this process requires the COOH-terminal coiled coil domain of SCP3, as well as the NH₂-terminal nonhelical domain. We have further analyzed the thick SCP3 fibers by transmission electron microscopy and immunoelectron microscopy. We found that the fibers display a transversal striation with a periodicity of ~20 nm and consist of a large number of closely associated, thin fibers, 5–10 nm in diameter. These features suggest that the SCP3 fibers are structurally related to intermediate filaments. It is known that in some species the lateral elements of the synaptonemal complex show a highly ordered striated structure resembling that of the SCP3 fibers. We propose that SCP3 fibers constitute the core of the lateral elements of the synaptonemal complex and function as a molecular framework to which other proteins attach, regulating DNA binding to the chromatid axis, sister chromatid cohesion, synapsis, and recombination.     

Nurse-Protocol Management of Low Back Pain: Outcomes,Patient Satisfaction and Efficiency of Primary Care

To test the validity of a nurse-administered protocol for low back pain, a prospective trial of 419 patients was undertaken in a walk-in clinic. In all, 222 patients were randomly allocated to a “nurse-protocol group” in which they were evaluated by one of five nurses using the protocol; the nurses independently managed 53 percent of the patients and referred to a physician patients with potentially complex conditions. In addition, 197 patients in a randomly allocated control group were managed by one of 32 physicians. Care in the experimental and control groups was compared by follow-up telephone contact and by a four-month chart review. There was no significant difference in symptomatic relief or the development of serious disease in the two groups. Nurse-protocol patients expressed greater satisfaction with the care they had received; patient satisfaction correlated positively with symptom relief. In over 95 percent of the patients, there were noncomplex, nonserious, nonchronic conditions as the cause of back pain. We conclude that nurse-protocol management of this generally benign condition in a primary care setting is both effective and efficient.     

Headwater Mires Constitute a Major Source of Nitrogen (N) to Surface Waters in the Boreal Landscape

Nutrient exports from soils have important implications for long-term patterns of nutrient limitation on land and resource delivery to aquatic environments. While plant–soil systems are notably efficient at retaining limiting nutrients, spatial and temporal mismatches in resource supply and demand may create opportunities for hydrologic losses to occur. Spatial mismatches may be particularly important in peat-forming landscapes, where the development of a two-layer vertical structure can isolate plant communities on the surface from resource pools that accumulate at depth. Our objectives were to test this idea in northern Sweden, where nitrogen (N) limitation of terrestrial plants is widespread, and where peat-forming, mire ecosystems are dominant features of the landscape. We quantified vertical patterns of N chemistry in a minerogenic mire, estimated the seasonal and annual hydrologic export of organic and inorganic N from this system, and evaluated the broader influence of mire cover on N chemistry across a stream network. Relatively high concentrations of ammonium (up to 2 mg l^?1) were observed in groundwater several meters below the peat surface, and N was routed to the outlet stream along deep, preferential flowpaths. Areal estimates of inorganic N export from the mire were several times greater than from an adjacent, forested catchment, with markedly higher loss rates during the growing season, when plant N demand is ostensibly greatest. At broader scales, mire cover was positively correlated with long-term concentrations of inorganic and organic N in streams across the drainage network. This study provides an example of how mire formation and peat accumulation can create broad-scale heterogeneity in nutrient supply and demand across boreal landscapes. This mismatch allows for hydrologic losses of reactive N that are independent of annual plant demand and potentially important to receiving lakes and streams.     

Nutrient Constraints on Metabolism Affect the Temperature Regulation of Aquatic Bacterial Growth Efficiency

Inorganic nutrient availability and temperature are recognized as major regulators of organic carbon processing by aquatic bacteria, but little is known about how these two factors interact to control bacterial metabolic processes. We manipulated the temperature of boreal humic stream water samples within 0–25°C and measured bacterial production (BP) and respiration (BR) with and without inorganic nitrogen?+?phosphorus addition. Both BP and BR increased exponentially with temperature in all experiments, with Q ₁₀ values varying between 1.2 and 2.4. The bacterial growth efficiency (BGE) showed strong negative relationships with temperature in nutrient-enriched samples and in natural stream water where community-level BP and BR were not limited by nutrients. However, there were no relationships between BGE and temperature in samples where BP and BR were significantly constrained by the inorganic nutrient availability. The results suggest that metabolic responses of aquatic bacterial communities to temperature variations can be strongly dependent on whether the bacterial metabolism is limited by inorganic nutrients or not. Such responses can have consequences for both the carbon flux through aquatic food webs and for the flux of CO₂ from aquatic systems to the atmosphere.     

A model system for increasing the intensity of whole-cell biocatalysis: investigation of the rate of oxidation of D-sorbitol to L-sorbose by thin bi-layer latex coatings of non-growing Gluconobacter oxydans

We developed a novel <50-microm thick nano-porous bi-layer latex coating for preserving Gluconobacter oxydans, a strict aerobe, as a whole cell biocatalyst. G. oxydans was entrapped in an acrylate/vinyl acetate co-polymer matrix (T (g) approximately 10 degrees C) and cast into 12.7-mm diameter patch coatings (cellcoat) containing approximately 10(9) CFU covered by a nano-porous topcoat. The oxidation of D-sorbitol to L-sorbose was used to investigate the coating catalytic properties. Intrinsic kinetics was studied in microbioreactors using a pH 6.0 D-sorbitol, phosphate, pyruvate (SPP) non-growth medium at 30 degrees C, and the Michaelis-Menten constants determined. By using a diffusion cell, cellcoat and topcoat diffusivities, optimized by arresting polymer particle coalescence by glycerol and/or sucrose addition, were determined. Cryo-FESEM images revealed a two-layer structure with G. oxydans surrounded by <40-nm pores. Viable cell density, cell leakage, and oxidation kinetics in SPP medium for >150 h were investigated. Even though the coatings were optimized for permeability, approximately 50% of G. oxydans viability was lost during cellcoat drying and further reduction was observed as the topcoat was added. High reaction rates per unit volume of coating (80-100 g/L x h) were observed which agreed with predictions of a diffusion-reaction model using parameters estimated by independent experiments. Cellcoat effectiveness factors of 0.22-0.49 were observed which are 20-fold greater than any previously reported for this G. oxydans oxidation. These nano-structured coatings and the possibility of improving their ability to preserve G. oxydans viability may be useful for engineering highly reactive adhesive coatings for multi-phase micro-channel and membrane bioreactors to dramatically increase the intensity of whole-cell oxidations.     

Caspase cleaved presenilin-1 is part of active gamma-secretase complexes

gamma-Secretase is a key enzyme involved in the processing of the beta-amyloid precursor protein into amyloid beta-peptides (Abeta). Abeta accumulates and forms plaques in Alzheimer's disease (AD) brains. A progressive neurodegeneration and cognitive decline occurs during the course of the disease, and Abeta is believed to be central for the molecular pathogenesis of AD. Apoptosis has been implicated as one of the mechanisms behind the neuronal cell loss seen in AD. We have studied preservation and activity of the gamma-secretase complex during apoptosis in neuroblastoma cells (SH-SY5Y) exposed to staurosporine (STS). We report that the known components (presenilin, Nicastrin, Aph-1 and Pen-2) interact and form active gamma-secretase complexes in apoptotic cells. In addition, the fragments corresponding to the PS1 N-terminal fragment and the caspase-cleaved PS1 C-terminal fragment (PS1-caspCTF) were found to form active gamma-secretase complexes when co-expressed in presenilin (PS) knockout cells. Interestingly, PS1-caspCTF replaced the normal PS1 C-terminal fragment and was co-immunoprecipitated with the gamma-secretase complex in SH-SY5Y cells exposed to STS. In addition, Abeta was detected in medium from apoptotic HEK APP(swe) cells. Together, the data show that gamma-secretase complexes containing PS1-caspCTF are active, and suggest that this proteolytic activity is also important in dying cells and may affect the progression of AD.     

Amino Acid Accumulation Limits the Overexpression of Proteins in Lactococcus lactis

Background

Understanding the biogenesis pathways for the functional expression of recombinant proteins, in particular membrane proteins and complex multidomain assemblies, is a fundamental issue in cell biology and of high importance for future progress in structural genomics. In this study, we employed a proteomic approach to understand the difference in expression levels for various multidomain membrane proteins in L. lactis cells grown in complex and synthetic media.

Methodology/Principal Findings

The proteomic profiles of cells growing in media in which the proteins were expressed to high or low levels suggested a limitation in the availability of branched-chain amino acids, more specifically a too limited capacity to accumulate these nutrients. By supplying the cells with an alternative path for accumulation of Ile, Leu and/or Val, i.e., a medium supplement of the appropriate dipeptides, or by engineering the transport capacity for branched-chain amino acids, the expression levels could be increased several fold.

Conclusions

We show that the availability of branched chain amino acids is a critical factor for the (over)expression of proteins in L. lactis. The forward engineering of cells for functional protein production required fine-tuning of co-expression of the branched chain amino acid transporter.