Climate change mitigation in Canada's forest sector: a spatially explicit case study for two regions.

BACKGROUND: We determine the potential of forests and the forest sector to mitigate greenhouse gas (GHG) emissions by changes in management practices and wood use for two regions within Canada's managed forest from 2018 to 2050. Our modeling frameworks include the Carbon Budget Model of the Canadian Forest Sector, a framework for harvested wood products that estimates emissions based on product half-life decay times, and an account of marginal emission substitution benefits from the changes in use of wood products and bioenergy. Using a spatially explicit forest inventory with 16 ha pixels, we examine mitigation scenarios relating to forest management and wood use: increased harvesting efficiency; residue management for bioenergy; reduced harvest; reduced slashburning, and more longer-lived wood products. The primary reason for the spatially explicit approach at this coarse resolution was to estimate transportation distances associated with delivering harvest residues for heat and/or electricity production for local communities. RESULTS: Results demonstrated large differences among alternative scenarios, and from alternative assumptions about substitution benefits for fossil fuel-based energy and products which changed scenario rankings. Combining forest management activities with a wood-use scenario that generated more longer-lived products had the highest mitigation potential. CONCLUSIONS: The use of harvest residues to meet local energy demands in place of burning fossil fuels was found to be an effective scenario to reduce GHG emissions, along with scenarios that increased the utilization level for harvest, and increased the longevity of wood products. Substitution benefits from avoiding fossil fuels or emissions-intensive products were dependent on local circumstances for energy demand and fuel mix, and the assumed wood use for products. As projected future demand for biomass use in national GHG mitigation strategies could exceed sustainable biomass supply, analyses such as this can help identify biomass sources that achieve the greatest mitigation benefits.

A large-diameter hollow-shaft cryogenic motor based on a superconducting magnetic bearing for millimeter-wave polarimetry.

In this paper, we present the design and measured performance of a novel cryogenic motor based on a superconducting magnetic bearing (SMB). The motor is tailored for use in millimeter-wave half-wave plate (HWP) polarimeters, where a HWP is rapidly rotated in front of a polarization analyzer or polarization-sensitive detector. This polarimetry technique is commonly used in cosmic microwave background polarization studies. The SMB we use is composed of fourteen yttrium barium copper oxide (YBCO) disks and a contiguous neodymium iron boron (NdFeB) ring magnet. The motor is a hollow-shaft motor because the HWP is ultimately installed in the rotor. The motor presented here has a 100 mm diameter rotor aperture. However, the design can be scaled up to rotor aperture diameters of approximately 500 mm. Our motor system is composed of four primary subsystems: (i) the rotor assembly, which includes the NdFeB ring magnet, (ii) the stator assembly, which includes the YBCO disks, (iii) an incremental encoder, and (iv) the drive electronics. While the YBCO is cooling through its superconducting transition, the rotor is held above the stator by a novel hold and release mechanism. The encoder subsystem consists of a custom-built encoder disk read out by two fiber optic readout sensors. For the demonstration described in this paper, we ran the motor at 50 K and tested rotation frequencies up to approximately 10 Hz. The feedback system was able to stabilize the rotation speed to approximately 0.4%, and the measured rotor orientation angle uncertainty is less than 0.15°. Lower temperature operation will require additional development activities, which we will discuss.

Historical effects of dissolved organic carbon export and land management decisions on the watershed-scale forest carbon budget of a coastal British Columbia Douglas-fir-dominated landscape.

BACKGROUND: To address how natural disturbance, forest harvest, and deforestation from reservoir creation affect landscape-level carbon (C) budgets, a retrospective C budget for the 8500 ha Sooke Lake Watershed (SLW) from 1911 to 2012 was developed using historical spatial inventory and disturbance data. To simulate forest C dynamics, data was input into a spatially-explicit version of the Carbon Budget Model-Canadian Forest Sector (CBM-CFS3). Transfers of terrestrial C to inland aquatic environments need to be considered to better capture the watershed scale C balance. Using dissolved organic C (DOC) and stream flow measurements from three SLW catchments, DOC load into the reservoir was derived for a 17-year period. C stocks and stock changes between a baseline and two alternative management scenarios were compared to understand the relative impact of successive reservoir expansions and sustained harvest activity over the 100-year period. RESULTS: Dissolved organic C flux for the three catchments ranged from 0.017 to 0.057 Mg C ha-1 year-1. Constraining CBM-CFS3 to observed DOC loads required parameterization of humified soil C losses of 2.5, 5.5, and 6.5%. Scaled to the watershed and assuming none of the exported terrestrial DOC was respired to CO2, we hypothesize that over 100 years up to 30,657 Mg C may have been available for sequestration in sediment. By 2012, deforestation due to reservoir creation/expansion resulted in the watershed forest lands sequestering 14 Mg C ha-1 less than without reservoir expansion. Sustained harvest activity had a substantially greater impact, reducing forest C stores by 93 Mg C ha-1 by 2012. However approximately half of the C exported as merchantable wood during logging (~176,000 Mg C) may remain in harvested wood products, reducing the cumulative impact of forestry activity from 93 to 71 Mg C ha-1. CONCLUSIONS: Dissolved organic C flux from temperate forest ecosystems is a small but persistent C flux which may have long term implications for C storage in inland aquatic systems. This is a first step integrating fluvial transport of C into a forest carbon model by parameterizing DOC flux from soil C pools. While deforestation related to successive reservoir expansions did impact the watershed-scale C budget, over multi-decadal time periods, sustained harvest activity was more influential.

Impact of adding Saccharomyces strains on fermentation, aerobic stability, nutritive value, and select lactobacilli populations in corn silage.

Bacterial inoculants can improve the conservation and nutritional quality of silages. Inclusion of the yeast Saccharomyces in the diet of dairy cattle has also been reported to be beneficial. The present study assessed the ability of silage to be used as a means of delivering Saccharomyces strains to ruminants. Two strains of Saccharomyces cerevisiae (strain 1 and 3)and 1 strain of Saccharomyces paradoxus (strain 2) were inoculated (10(3) cfu/g) individually onto corn forage that was ensiled in mini silos for 90 d. Fermentation characteristics, aerobic stability, and nutritive value of silages were determined and real-time quantitative PCR (RT-qPCR) was used to quantify S. cerevisiae, S.paradoxus, total Saccharomyces, fungal, and bacterial populations. Fermentation characteristics of silage inoculated with S1 were similar to control silage. Although strain 3 inoculation increased ash and decreased OM contents of silage (P = 0.017), no differences were observed in nutrient composition or fermentation profiles after 90 d of ensiling. Inoculation with Saccharomyces had no detrimental effect on the aerobic stability of silage. In vitro DM disappearance, gas production, and microbial protein synthesis were not affected by yeast inoculation.Saccharomyces strain 1 was quantified throughout ensiling, whereas strain 2 was detected only immediately after inoculation. Saccharomyces cerevisiae strain 3 was quantified until d 7 and detectable 90 d after ensiling. All inoculants were detected and quantified during aerobic exposure. Inoculation with Saccharomyces did not alter lactobacilli populations. Saccharomycetales were detected by RT-qPCR throughout ensiling in all silages. Both S. cerevisiae and S. paradoxus populations increased during aerobic exposure, demonstrating that the density of these yeast strains would increase between the time that silage was removed from storage and the time it was fed.

Comparative denaturing gradient gel electrophoresis analysis of fungal communities associated with whole plant corn silage.

Significant portions of grain produced for livestock consumption are convened into ensiled forage. Silage producers have long recognized the positive effects of using an inoculant to insure the proper transformation of forage into a palatable and digestible feedstuff. When silage is fed from a storage structure, exposure to air stimulates the growth of epiphytic aerobes that may result in the loss of up to 50% of the dry matter. Moreover, fungi have been found to be associated with ensiled forage, but their growth is normally suppressed by the anaerobic conditions. However, the introduction of oxygen results in a fungal bloom, and the fungi and the associated metabolites may result in lost productivity in the livestock consuming the contaminated forage. In this study, we report on the diversity of the fungal community associated with whole plant corn silage during the ensiling process, and the effect of two different bacterial inoculants as compared with the uninoculated natural epiphytic fermentation on the distribution of the fungi associated with the silage. The fungal community from duplicate mini-silo packages of the same treatment was analyzed by denaturing gradient gel electrophoresis and direct sequencing of the resulting operational taxonomic units. This method proved useful in analyzing the complex microbial communities associated with the forage in that it was possible to determine that one inoculant dramatically influenced the fungal community associated with whole plant corn silage.

Assembly of long chain phosphatidylcholines at a liquid-liquid interface.

The molecular-level organization of mixed and pure saturated symmetric chain 1,2-diacyl-sn-glycero-3-phosphocholines (PCs) adsorbed at a carbon tetrachloride-aqueous interface is explored by probing the hydrocarbon chain conformation within the adsorbed layer. PCs of the chain lengths found most frequently in biological systems, which in pure form are seen to form either very well-ordered or disordered layers, are observed in these studies to assemble into interfacial layers ranging from disordered to ordered states when mixed in various proportions. Independently, while C(16) and shorter chain PCs tend to form disordered layers, a strong increase in ordering is observed for C(18) and longer chain PCs in which the hydrocarbon chains are found to be primarily in an all trans conformation. Pure C(17)-PCs adsorbed at the interface produce layers with an intermediate degree of chain ordering. The ability to tune interfacial layer properties in mixed systems as a function of molecular composition, including PC chain length as demonstrated here, is an important mechanism by which surface characteristics of oil-water emulsion systems can be controlled both in vivo and in numerous commercial applications.

Use of an enhanced Escherichia coli opal suppressor strain to screen a Mycoplasma hyopneumoniae library.

Many mycoplasma genes contain internal UGA (opal) codons because of their use as tryptophan coding codons. This results in a lack of expression of many cloned mycoplasma antigenic epitopes in Escherichia coli. It has been shown that opal suppressors can be used to enhance expression of defined mycoplasma gene sequences, but no studies have been published using E. coli suppressor strains to screen mycoplasma gene libraries for immunoreactive epitopes. The E. coli suppressor strain ISM612 was used to screen a Mycoplasma hyopneumoniae Lambda gene library. This strain contained an inducible opal suppressor, trpT, as well as the release factor 2 mutation prfB3. Strain ISM612 was shown to enhance antibody recognition of cloned mycoplasmal gene sequences.

A genetic and molecular characterization of the recA gene from Staphylococcus aureus.

Previous studies have identified mutant strains of Staphylococcus aureus that have deficiencies in genetic recombination and DNA repair. Although these phenotypes were tentatively attributed to mutations within the S. aureus recA gene, experimental evidence to confirm this has never been reported. To characterize recA from S. aureus, we first isolated transposon insertion mutations that were in close proximity to the recA-like mutation (uvs-568) in strain 112 UVS-1. This allowed for the mobilization of the uvs-568 mutation into strain RN4220, the common laboratory strain of S. aureus. Next, using Bacillus subtilis recA as a probe, we cloned S. aureus recA and determined its nucleotide sequence. The deduced amino acid (aa) sequence of RecA contained 347 aa and was 74% identical to B. subtilis RecA. Using a cloned DNA fragment originating from within S. aureus recA, we then constructed a recA null mutant strain, designated KB103, which exhibited the same phenotypic characteristics imposed by the uvs-568 mutation in the same background. Furthermore, genetic and physical mapping of S. aureus recA placed it in the same region as the uvs-568 mutation. These data strongly suggest that these mutations represent different alleles of the same recA gene.

Expression of a retroposon-like sequence upstream of the putative Trypanosoma brucei variant surface glycoprotein gene expression site promoter.

We have cloned the region spanning the putative promoter from two variant surface glycoprotein gene expression sites that are at each end of chromosome M4 of Trypanosoma brucei IsTat 7. Both expression sites contain a retroposon-like sequence (ESR) pseudogene whose 3' end is approximately 30 bp upstream of the putative expression site promoter. The ESRs from both expression sites share considerable sequence homology and are related to LINE-like elements, especially the T. brucei ingi retroposon. Other ESRs are located on large, but not intermediate or mini-, chromosomes in the IsTaR 1 serodeme, and the total copy number is 10 to 20, similar to that estimated for variant surface glycoprotein expression sites. No DNA rearrangements in the vicinity of the ESR and putative expression site promoter were detected following antigenic switches in the IsTaR 1 serodeme. ESR transcripts are present in bloodstream, but not procyclic, forms. Variation in transcript size and sequence between bloodstream variant antigenic types implies that only the ESR from the active expression site is transcribed. This pattern of expression reflects that of sequences downstream of the putative expression site promoter, suggesting that the region of coordinately controlled expression extends upstream of this promoter.

Enhanced readthrough of opal (UGA) stop codons and production of Mycoplasma pneumoniae P1 epitopes in Escherichia coli.

Expression of mycoplasma sequences in Escherichia coli is often hindered by an unusual mycoplasmal codon usage pattern: the UGA stop codon is utilized for tryptophan. This may result in the truncation of cloned proteins and may prevent the detection of products of many cloned genes. To circumvent this translation barrier, we have developed an expression system for the production of mycoplasma proteins in E. coli. The efficiency of an opal suppressor tRNA (trpT176) was augmented with other suppressor mutations (prfB3 or rrsB(SuUGA-delta C1054)) which influence termination events. System efficacy was analyzed by employing suppressor mutations in the expression of TGA-containing sequences from the P1 protein-encoding gene of Mycoplasma pneumoniae.

Insertion of Tn916 into Bacillus pumilus plasmid pMGD302 and evidence for plasmid transfer by conjugation.

As part of an effort to develop systems for genetic analysis of strains of Bacillus pumilus which are being used as a microbial hay preservative, we introduced the conjugative Enterococcus faecalis transposon Tn916 into B. pumilus ATCC 1 and two naturally occurring hay isolates of B. pumilus. B. pumilus transconjugants resistant to tetracycline were detected at a frequency of approximately 6.5 x 10(-7) per recipient after filter mating with E. faecalis CG110. Southern hybridization confirmed the insertion of Tn916 into several different sites in the B. pumilus chromosome. Transfer of Tn916 also was observed between strains of B. pumilus in filter matings, and one donor strain transferred tetracycline resistance to recipients in broth matings at high frequency (up to 3.4 x 10(-5) per recipient). Transfer from this donor strain in broth matings was DNase-resistant and was not mediated by culture filtrates. Transconjugants from these broth matings contained derivatives of a cryptic plasmid (pMGD302, approx 60 kb) from the donor strain with Tn916 inserted at various sites. The plasmids containing Tn916 insertions transferred to a B. pumilus recipient strain at frequencies of approx 5 x 10(-6) per recipient. This evidence suggests that pMGD302 can transfer by a process resembling conjugation between strains of B. pumilus.

Hepatic protein tyrosine phosphatases in the rat.

Regulation of cell growth and metabolism by protein tyrosine phosphorylation involves dephosphorylation via the action of protein tyrosine phosphatases (PTPases). We have characterized the membrane PTPases in rat liver, monitoring their activity by measuring the dephosphorylation of P-Tyr-reduced, carboxyamidomethylated and maleylated lysozyme (P-Tyr-RCML) and P-Tyr-myelin basic protein (P-Tyr-MBP). Separation of membrane PTPases by poly (L-lysine) chromatography yielded three peaks of PTPase, termed I, II and III. PTPases I and II were most active with P-Tyr-RCML, whereas PTPase III showed greater activity with P-Tyr-MBP than with P-Tyr-RCML (ratio of activities 4:1). Separation of membrane proteins by gel-filtration chromatography yielded two peaks of activity. Based on substrate specificity, sensitivity to inhibitors and requirement for thiol-containing compounds, the activity peak with an Mr of approximately 400,000 corresponded to PTPase III, whereas that with an Mr of approx. 40,000 contained PTPases I and II. All three PTPases dephosphorylated epidermal growth factor receptors and insulin receptors, but only PTPases I and II were active with P-Tyr-asialoglycoprotein receptors. Although none of the above characteristics distinguished between PTPases I and II, only PTPase I reacted in a Western immunoblotting procedure with anti-peptide antibodies directed towards human placental PTPase. We conclude that the membrane fraction from rat liver contains at least three distinct PTPases.

Does the duration of N2O administration affect postoperative nausea and vomiting?

Nausea and vomiting are the most frequent postoperative complications in the ambulatory surgical setting. In the present study, data were obtained from 184 adult ambulatory cosmetic surgery patients to determine if the use of nitrous oxide (N2O) was associated with an increased incidence of postoperative nausea and/or vomiting (PNV). Anesthesia was induced with thiopental and maintained with an opioid (fentanyl or sufentanil) and isoflurane with or without N2O. Data were analyzed statistically using the two-way chi-square test and the Fisher Exact Test. The major finding was that the incidence of PNV was directly related to the duration of anesthesia in the patients who received N2O, but not in those who were N2O-free. Anesthesia times and the percentages of patients who exhibited PNV in the N2O-free and N2O-treated groups, respectively, were: (1) less than 1 hour, 0% and 6.3%; (2) between 1.0 and 1.9 hours, 35.3% and 36.8%; (3) between 2.0 and 2.9 hours, 24.2% and 66.7% (p = .06); (4) between 3.0 and 5.3 hours, 35% and 100% (P less than .05). Thus, avoidance of N2O in ambulatory cosmetic surgery cases lasting 3, and quite probably 2 or more hours in which general anesthesia is maintained with a synthetic opioid and isoflurane appears to reduce the likelihood that these short-stay patients will experience PNV.

Use of a Plasmid DNA Probe To Monitor Populations of Bacillus pumilus Inoculant Strains in Hay.

We are evaluating naturally occurring isolates of Bacillus pumilus for use as microbial hay preservatives. Seven isolates of B. pumilus from hay contained a 42-kb cryptic plasmid (pMGD296). We wished to determine whether pMGD296 could be used as a molecular marker to follow populations of these isolates in hay over time. Southern blots and colony blots of 69 isolates of B. pumilus and other Bacillus spp. were probed with P-labeled pMGD296. Twenty-nine probe-positive isolates were identified; of these, 28 contained a plasmid with a restriction profile identical to that of pMGD296. One isolate from untreated hay contained a 40-kb plasmid (pMGD150) that was homologous to pMGD296 but had a different restriction fragment pattern. Regions of homology between the two plasmids were identified by Southern blotting, and a 1.9-kb HindIII-PstI fragment of pMGD296 lacking strong homology to pMGD150 was cloned in pUC18. The cloned fragment hybridized only with isolates containing pMGD296 and was used to estimate populations of these isolates in treated and untreated hay.

The trypanosome leucine repeat gene in the variant surface glycoprotein expression site encodes a putative metal-binding domain and a region resembling protein-binding domains of yeast, Drosophila, and mammalian proteins.

We have identified a new variant surface glycoprotein expression site-associated gene (ESAG) in Trypanosoma brucei, the trypanosome leucine repeat (T-LR) gene. Like most other ESAGs, it is expressed in a life cycle stage-specific manner. The N-terminal 20% of the predicted T-LR protein resembles the metal-binding domains of nucleic acid-binding proteins. The remainder is composed of leucine-rich repeats that are characteristic of protein-binding domains found in a variety of other eucaryote proteins. This is the first report of leucine-rich repeats and potential nucleic acid-binding domains on the same protein. The T-LR gene is adjacent to ESAG 4, which has homology to the catalytic domain of adenylate cyclase. This is intriguing, since yeast adenylate cyclase has a leucine-rich repeat regulatory domain. The leucine-rich repeat and putative metal-binding domains suggest a possible regulatory role that may involve adenylate cyclase activity or nucleic acid binding.

A retroposon in the 5' flank of a Trypanosoma brucei VSG gene lacks insertional terminal repeats.

A retroposon-like repeated sequence, ingi, occurs in high copy number in the genome of Trypanosoma brucei brucei. An ingi is present in the 5' flank of the 5C gene, an intrachromosomal IsTat 1.5 variant surface glycoprotein (VSG) gene family member. The 5' end of the ingi is located 22 bp upstream of the putative VSG start codon and the ingi open reading frame is in the opposite orientation to that of the VSG gene. The termini of the ingi are not flanked by a short repeat sequence and there are no sequences upstream of the ingi insertion which are homologous to the 5' flanking sequence of other 5 VSG gene family members. Thus, it appears that recombination and/or gene conversion between two ingi sequences may have eliminated the original 5C gene flanking sequence. Similar events may also have occurred with all but one previously reported ingi.

An extensively edited mitochondrial transcript in kinetoplastids encodes a protein homologous to ATPase subunit 6.

The mitochondrial MURF4 gene of T. brucei has pronounced G versus C strand bias and heterogeneously sized transcripts, characteristic of genes encoding extensively edited transcripts. We find that MURF4 transcripts of T. brucei are extensively edited throughout by the addition and deletion of numerous uridines, creating potential initiation and termination codons and a continuous open reading frame. A potential guide RNA sequence occurs in a minicircle between inverted repeats. The 5' region of L. tarentolae MURF4 transcripts is also extensively edited, with a created initiation codon. The predicted MURF4 amino acid sequences have homology to those of mitochondrial ATP-ase subunit 6 genes from a variety of organisms. In addition, their hydropathic profiles are quite similar to those of other species. We therefore conclude that MURF4 encodes ATPase subunit 6 genes.