Variation in leaf physiology of <Emphasis Type="Italic">Salix arctica</Emphasis> within and across ecosystems in the High Arctic: test of a dual isotope (Δ<Superscript>13</Superscript>C and Δ<Superscript>18</Superscript>O) conceptual model

Leaf carbon isotope discrimination (Δ¹³C) varies with the balance between net photosynthesis (A) and stomatal conductance (g _s ). Inferences that can be made with Δ¹³C are limited, as changes could reflect variation in A and/or g _s . Investigators have suggested that leaf δ¹⁸O enrichment above source water (Δ¹⁸O) may enable differentiation between sources of variation in Δ¹³C, as leaf Δ¹⁸O varies with transpiration rate (E), which is closely correlated with g _s when leaves experience similar leaf to air vapor pressure differences. We examined leaf gas exchange of Salix arctica at eight sites with similar air temperatures and relative humidities but divergent soil temperatures and soil water contents near Pituffik, Greenland (76°N, 38°W). We found negative correlations at the site level between g _s and Δ¹⁸O in bulk leaf tissue (r ² = 0.62, slope = −17.9‰/mol H₂O m⁻² s⁻¹, P = 0.02) and leaf α-cellulose (r ² = 0.83, slope = −11.5‰ mol H₂O m⁻² s⁻¹, P < 0.01), consistent with the notion that leaf water enrichment declines with increasing E. We also found negative correlations at the site-level between intrinsic water-use efficiency (iWUE) and Δ¹³C in bulk leaf tissue (r ² = 0.65, slope = −0.08‰/μmol CO₂ /mol H₂O, P = 0.02) and leaf α-cellulose (r ² = 0.50, slope = −0.05 ‰/[μmol CO₂ /mol H₂O], P = 0.05). When increasing Δ¹³C was driven by increasing g _s alone, we found negative slopes between Δ¹³C and Δ¹⁸O for bulk leaf tissue (−0.664) and leaf α-cellulose (−1.135). When both g _s and A _max increased, we found steeper negative slopes between Δ¹³C and Δ¹⁸O for bulk leaf tissue (−2.307) and leaf α-cellulose (−1.296). Our results suggest that the dual isotope approach is capable of revealing the qualitative contributions of g _s and A _max to Δ¹³C at the site level. In our study, bulk leaf tissue was a better medium than leaf α-cellulose for application of the dual isotope approach.     

An update on the routine application of MALDI-TOF MS in clinical microbiology

Introduction: Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has entered clinical diagnostics and is today a generally accepted and integral part of the workflow for microbial identification. MALDI-TOF MS identification systems received approval from national and international institutions, such as the USA-FDA, and are continuously improved and adopted to other fields like veterinary and industrial microbiology. The question is whether MALDI-TOF MS also has the potential to replace other conventional and molecular techniques operated in routine diagnostic laboratories.

Areas covered: We give an overview of new advancements of mass spectral analysis in the context of microbial diagnostics. In particular, the expansion of databases to increase the range of readily identifiable bacteria and fungi, the refined discrimination of species complexes, subspecies, and types, the testing for antibiotic resistance or susceptibility, progress in sample preparation including automation, and applications of other mass spectrometry techniques are discussed.

Expert opinion: Although many new approaches of MALDI-TOF MS are still in the stage of proof of principle, it is expectable that MALDI-TOF MS will expand its role in the clinical microbiology laboratory of the future. New databases, instruments and analytical software modules will continue to be developed to further improve diagnostic efficacy.     

Cooperative assembly of simian virus 40 T-antigen hexamers on functional halves of the replication origin.

The cofactor ATP stimulates the formation of T-antigen double hexamers on the simian virus 40 core origin of replication (I. A. Mastrangelo, P. V. C. Hough, J. S. Wall, M. Dodson, F. B. Dean, and J. Horwitz, Nature [London] 338:658-662, 1989). We report here the pathway for the assembly of hexamers and double hexamers on the core origin. ATP triggers the cooperative assembly of hexamers on the early and late halves of the origin even when they are completely isolated. Hexamer assembly nucleates at T-antigen recognition pentanucleotides in the early half of the origin. In intact origins, assembly of the first hexamer on the early half of the origin cooperatively stimulates the assembly of a second hexamer on the adjacent late half of the origin. Thus, monomer-monomer and hexamer-hexamer interactions of T antigen, allosterically activated by ATP, constitute two distinct types of cooperative interaction with the origin. Finally, we show that the assembly of T-antigen hexamers on isolated half origins leads to the same array of structural changes that T antigen induces in intact origins. We conclude that the origin is divided into complementary halves that each promote the assembly of functional T-antigen hexamers.     

Intercellular adhesion molecule-1 and hair follicle regression.

Although the intercellular adhesion molecule-1 (ICAM-1) is recognized for its pivotal role in inflammation and immune responses, its role in developmental systems, such as the cyclic growth (anagen) and regression (catagen) of the hair follicle, remains to be explored. Here we demonstrate that ICAM-1 expression in murine skin is even more widespread and more developmentally regulated than was previously believed. In addition to endothelial cells, selected epidermal and follicular keratinocyte subpopulations, as well as interfollicular fibroblasts, express ICAM-1. Murine hair follicles express ICAM-1 only late during morphogenesis. Thereafter, morphologically identical follicles markedly differ in their ICAM-1 expression patterns, which become strikingly hair cycle-dependent in both intra- and extrafollicular skin compartments. Minimal ICAM-1 and leukocyte function-associated (LFA-1) protein and mRNA expression is observed during early anagen and maximal expression during late anagen and catagen. Keratinocytes of the distal outer root sheath, fibroblasts of the perifollicular connective tissue sheath, and perifollicular blood vessels exhibit maximal ICAM-1 immunoreactivity during catagen, which corresponds to changes of LFA-1 expression on perifollicular macrophages. Finally, ICAM-1-deficient mice display significant catagen acceleration compared to wild-type controls. Therefore, ICAM-1 upregulation is not limited to pathological situations but is also important for skin and hair follicle remodeling. Collectively, this suggests a new and apparently nonimmunological function for ICAM-1-related signaling in cutaneous biology.     

Carbon import among vegetative tillers within two bunchgrasses: assessment with carbon-11 labelling

Summary Carbon allocation among bunchgrass tillers was examined with carbon-11 (¹¹CO₂) steady state labelling. Labelled carbon was continuously transported from parent tillers to anatomically attached daughter tillers at a time when morphological characteristics indicated that tiller maturation had occurred. Steady state levels of import into monitored daughter tillers increased within 30 min of either defoliation or shading. Import levels decreased within 30 min of the removal of shading, but remained accelerated throughout an 84 h observation period following defoliation. A second defoliation further increased carbon import into a monitored tiller above the previously accelerated level resulting from the initial defoliation. Carbon import by vegetative tillers in the two bunchgrass species examined may be most appropriately viewed as a series of potentially accelerated import levels above a low level of continuous import.     

A convenient incorporation of conformationally constrained 5,5-dimethylproline into the ribonuclease A 89-124 sequence by condensation of synthetic peptide fragments.

The presence of l-5,5-dimethylproline (dmP) within an amino acid sequence results in the formation of an X-dmP peptide bond predominantly locked in a cis conformation. However, the common use of this unnatural amino acid has been hampered by the difficulty of the economical incorporation of the dmP residue into longer peptide segments due to the steric hindrance imposed by the dimethyl moieties. Here, we describe synthesis of the C-terminal 36-residue peptide, corresponding to the 89-124 sequence of bovine pancreatic ribonuclease A (RNase A), in which dmP is incorporated as a substitute for Pro93. The peptide was assembled by condensation of protected 5- and 31-residue peptide fragments, which were synthesized by solid-phase peptide methodology using fluorenylmethyloxycarbonyl chemistry. We focused on optimizing the synthesis of the Fmoc-Ser(tBu)-Ser(tBu)-Lys(Boc)-Tyr(tBu)-dmP-OH pentapeptide (residues 89-93) with efficient acylation of the sterically hindered dmP residue. In a comparative study, the reagent O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate was found to be superior to bromo-tris-pyrrolidino-phosphonium hexafluorophosphate and tetramethylfluoroformamidinium hexafluorophosphate for the synthesis of the -Tyr(tBu)-dmP- peptide bond in solution as well as on a resin.     

Modeling the seasonality of belowground respiration along an elevation gradient in the western Chugach Mountains, Alaska

Belowground respiration is typically the largest flux of carbon from terrestrial ecosystems to the atmosphere, making up >70% of total respiration in boreal forests. Recent work has shown that belowground respiration continues during the snow-covered season in boreal ecosystems, but few studies have made complementary measurements during the snow-free season and it remains uncertain what proportion of annual belowground respiration occurs during winter. Traditional models of the relationship between temperature and respiration assume fixed temperature sensitivity, but it has become clear that the apparent temperature sensitivity of belowground respiration increases as soils approach 0°C. Use of fixed temperature sensitivity to model carbon budgets of northern ecosystems may, therefore, yield misleading results. We measured belowground respiration monthly over 2 years in four ecosystems along an elevation gradient in south-central Alaska. Three models, representing different hypotheses about the relationship between temperature and respiration, were confronted with the data. A logistic model, which allows the temperature sensitivity to vary inversely with temperature, and a variation of the Q₁₀ model, which allows the temperature sensitivity to vary seasonally, performed well at all sites and produced similar estimates of seasonal and annual belowground respiration. The traditional Q₁₀ model performed poorly at all sites and overestimated respiration during the snow-covered season. Annual belowground respiration was generally greater than in ecosystems of interior Alaska, where winters are colder and summers are warmer and drier. Belowground respiration during the snow-covered season made up 6–15% of the annual total—a small, but sensitive, component of annual carbon budgets.     

SNL fibroblast feeder layers support derivation and maintenance of human induced pluripotent stem cells

Induced pluripotent stem(iPS) cells can be derived from human somatic cells by cellular reprogramming.This technology provides a potential source of non-controversial therapeutic cells for tissue repair,drug discovery,and opportunities for studying the molecular basis of human disease.Normally,mouse embryonic fibroblasts(MEFs) are used as feeder layers in the initial derivation of iPS lines.The purpose of this study was to determine whether SNL fibroblasts can be used to support the growth of human iPS cell...     

Prevalence and molecular characterization of Mycoplasma ovis in selected free-ranging Brazilian deer populations

Mycoplasma ovis is a hemoplasma that may cause anemia and mortality in small ruminants. Our aim was to determine whether M. ovis infects populations of free-ranging deer in Brazil. Buffy coat samples from 64 Blastocerus dichotomus from Porto Primavera, 18 Ozotocerus bezoarticus from Pantanal, and 21 O. bezoarticus from Emas National Park were tested. Using a M. ovis PCR protocol to amplify extracted DNA, 46/64 (72%) of deer from Porto Primavera, 10/18 (56%) from Pantanal, and 4/21 (19%) from Emas National Park were positive, giving an overall positive rate of 58% for hemoplasma in these wild deer. Sequencing and phylogenetic analysis of the 16S rRNA gene revealed 3 genetically distinct hemoplasmas including M. ovis, 'Candidatus Mycoplasma erythrocervae', and a hemoplasma most closely related to M. ovis. Phylogenetic analysis of the 23S rRNA gene from selected sequences confirmed these relationships.     

Soil–plant N processes in a High Arctic ecosystem,NW Greenland are altered by long‐term experimental warming and higher rainfall

Rapid temperature and precipitation changes in High Arctic tundra ecosystems are altering the biogeochemical cycles of carbon (C) and nitrogen (N), but in ways that are difficult to predict. The challenge grows from the uncertainty of N cycle responses and the extent to which shifts in soil N are coupled with the C cycle and productivity of tundra systems. We used a long‐term (since 2003) experiment of summer warming and supplemental summer water additions to a High Arctic ecosystem in NW Greenland, and applied a combination of discrete sampling and in situ soil core incubations to measure C and N pools and seasonal microbial processes that might control plant‐available N. We hypothesized that elevated temperature and increased precipitation would stimulate microbial activity and net inorganic N mineralization, thereby increasing plant N‐availability through the growing season. While we did find increased N mineralization rates under both global change scenarios, water addition also significantly increased net nitrification rates, loss of NO₃^?‐N via leaching, and lowered rates of labile organic N production. We also expected the chronic warming and watering would lead to long‐term changes in soil N‐cycling that would be reflected in soil δ¹⁵N values. We found that soil δ¹⁵N decreased under the different climate change scenarios. Our results suggest that temperature accelerates biological processes and existing C and N transformations, but moisture increases soil hydraulic connectivity and so alters the pathways, and changes the fate of the products of C and N transformations. In addition, our findings indicate that warmer, wetter High Arctic tundra will be cycling N and C in ways that may transform these landscapes in part leading to greater C sequestration, but simultaneously, N losses from the upper soil profile that may be transported to depth dissolved in water and or transported off site in lateral flow.