Activated human T cells can present alloantigens but cannot present soluble antigens

Human T cells, when activated by antigen or mitogen, express Ia antigens. We have examined the capacity of activated T cells to stimulate autologous and allogeneic T cells and their ability to present soluble antigen. Interleukin 2-dependent T-cell lines (TCL), free of accessory cells, were used for antigen-presenting cells. These activated T cells were potent stimulators in an autologous mixed lymphocyte reaction (AMLR), more so than autologous irradiated non-T mononuclear cells. Activated T cells were also able to stimulate proliferation of allogeneic T cells in the absence of any other accessory cells, and this stimulation was blocked by anti-Ia antibodies. Resting unstimulated T cells were unable to stimulate autologous or allogeneic responses. Thus, activated T cells were able to present self antigens and alloantigens. However, activated T cells could not present soluble antigens to autologous T cells or to antigen-specific TCL even if exogenous interleukin 1 was added to cultures. The ability of activated T cells to stimulate an AMLR in vitro may reflect an important immunologic amplification mechanism in vivo. The ability of activated T cells to present alloantigens but not soluble antigens suggests an inability to process antigen, and this may provide further insights into the complexities of antigen presentation.     

On the mechanism of activation of the ATP X Mg(II)-dependent phosphoprotein phosphatase by kinase FA

The mechanism of activation of the Mg(II) X ATP-dependent phosphatase by the kinase FA has been investigated. The inactive protein phosphatase can be represented as FC X M where FC is the inactive catalytic component and M is the heat-stable modulator protein (also known as inhibitor-2). Phosphorylation of the modulator protein is demonstrated during activation of FC X M. In addition, continuous ATP hydrolysis during the activation is observed. This suggests that a cyclic phosphorylation-dephosphorylation reaction is continuously occurring during the activation. It is proposed that phosphorylation of the modulator protein causes an isomerization in FC to generate an active phosphatase. The activated phosphatase is capable of dephosphorylating the phosphorylated modulator. Upon dephosphorylation of modulator, the active phosphatase returns to its inactive form via a slow isomerization.     

Investigation of the pH dependence of proton uptake by porcine heart mitochondrial malate dehydrogenase upon binding of NADH

The pH dependence of proton uptake upon binding of NADH to porcine heart mitochondrial malate dehydrogenase (l-malate: NAD⁺ oxidoreductase, EC 1.1.1.37) has been investigated. The enzyme has been shown to exhibit a pH-dependent uptake of protons upon binding NADH at pH values from 6.0 to 8.5. Enzyme in which one histidine residue has been modified per subunit by the reagent iodoacetamide (E. M. Gregory, M. S. Rohrbach, and J. H. Harrison, 1971, Biochim. Biophys. Acta253, 489–497) was used to establish that this specific histidine residue was responsible for the uptake of a proton upon binding of NADH to the native enzyme. It has also been established that while there is no enhancement of the nucleotide fluorescence upon addition of NADH to the iodoacetamide-modified enzyme, NADH is nevertheless binding to the modified enzyme with the same stoichiometry as with native enzyme. The data are discussed in relation to the involvement of the essential histidine residue in the catalytic mechanism of “histidine dehydrogenases” recently proposed by Lodola et al. (A. Lodola, D. M. Parker, R. Jeck, and J. J. Holbrook, 1978, Biochem. J.173, 597–605) and the catalytic mechanism of “malate dehydrogenases” recently proposed by L. H. Bernstein and J. Everse (1978, J. Biol. Chem.253, 8702–8707).     

Impacts of annual weather conditions on forest productivity

The response of four northern deciduous tree species to annual climate variation is quantified at two intensively measured sites in northern Michigan, USA. Response to changes in temperature and moisture differ with the species and is dependent on other site conditions. Relationships identified in these field studies indicate that projected climate changes may have dramatic effects on the productivity of at least some commercially important tree species in the northern United States.     

Phosphorus Translocation by Red Deer on a Subalpine Grassland in the Central European Alps

We examined the role of red deer (Cervus elaphus L.) in translocating phosphorus (P) from their preferred grazing sites (short-grass vegetation on subalpine grasslands) to their wider home range in a subalpine grassland ecosystem in the Central European Alps. Phosphorus was used because it is the limiting nutrient in these grasslands. When we compared P removal of aboveground biomass due to grazing with P input due to the deposit of feces on a grid of 268 cells (20 m × 20 m) covering the entire grassland, we detected distinct spatial patterns: the proportion of heavily grazed short-grass vegetation increased with increasing soil-P pool, suggesting that red deer preferably grazed on grid cells with a higher soil-P pool. Biomass consumption related to increased proportion of short-grass vegetation, and therefore P removal, increased with increasing soil-P pool. However, within the two vegetation types (short-grass and tall-grass), consumption was independent from soil-P pool. In addition, P input rates from defecation increased with increasing soil-P pool, resulting in a constant mean net P loss of 0.083 kg ha⁻¹ y⁻¹ (0.03%–0.07% of soil-P pool) independent of both soil-P pool and vegetation type. Thus, there was no P translocation between grid cells with different soil-P pools or between short-grass and tall-grass vegetation. Based on these results, it is likely that the net rate of P loss is too small to explain the observed changes in vegetation composition from tall-herb/meadow communities to short-grass and from tall-grass to short-grass on the grassland since 1917. Instead, we suggest that the grazing patterns of red deer directly induced succession from tall-herb/meadow communities to short-grass vegetation. Yet, it is also possible that long-term net soil-P losses indirectly drive plant succession from short-grass to tall-grass vegetation, because nutrient depletion could reduce grazing pressure in short-grass vegetation and enable the characteristic tall-grass species Carex sempervirens Vill. to establish.     

Evidence of damage from exotic invasive earthworm activity was highly correlated to sugar maple dieback in the Upper Great Lakes region

Sugar maple (Acer sacharrum Marsh.) in the western Upper Great Lakes region has recently been reported with increased crown dieback symptoms, prompting investigation of the dieback etiology across the region. Evaluation of sugar maple dieback from 2009 to 2012 across a 120 plot network in Upper Michigan, northern Wisconsin, and eastern Minnesota has indicated that forest floor disturbance impacts from exotic invasive earthworms was significantly related to maple dieback. Other plot level variables tested showed significant relationships among dieback and increased soil carbon, decreased soil manganese, and reduced herbaceous cover, each of which was also be correlated to earthworm activity. Relationships between possible causal factors and recent growth trends and seedling counts were also examined. Maple regeneration counts were not correlated with the amount of dieback. The recent mean radial increment was significantly correlated with various soil features and nutrients. This study presents significant evidence correlating sugar maple dieback in the western Upper Great Lakes region with earthworm activity, and highlights the need for considering the impacts of non-native earthworm on soil properties when assessing sugar maple health and productivity.     

Kinase FA-mediated regulation of rabbit skeletal muscle protein phosphatase. Reversible phosphorylation of the modulator subunit

A mechanism of activation of the ATP.Mg-dependent protein phosphatase (FC.M) has been proposed (Jurgensen, S., Shacter, E., Huang, C. Y., Chock, P. B., Yang, S.-D., Vandenheede, J. R., and Merlevede, W. (1984) J. Biol. Chem. 259, 5864-5870) in which a transient phosphorylation by the kinase FA of the modulator subunit (M) is the driving force for the transition of the inactive catalytic subunit (FC) into its active conformation. Incubation of FC.M with kinase FA and Mg2+ and adenosine 5'-(gamma-thio)triphosphate results in thiophosphorylation of M and also a conformational change in the phosphatase catalytic subunit; however, the enzyme remains inactive. Proteolysis of this inactive, thiophosphorylated complex causes proteolytic destruction of the modulator subunit and yields an active phosphorylase phosphatase species. Similar treatment of the native inactive enzyme does not yield active phosphatase. Evidence is presented, suggesting that a molecule of modulator is bound at an "inhibitory site" on the native enzyme. This modulator does not prevent the conformational change in the phosphatase catalytic subunit upon incubation with kinase FA and ATP.Mg but does partially inhibit the expression of the phosphorylase phosphatase activity.     

Comparison of phenotypic and genetic clone delineation in quaking aspen, <Emphasis Type="Italic">Populus tremuloides</Emphasis>

Key message

Clonal delineation at nuclear microsatellites and phenotypic traits showed high correspondence and revealed an important role of both sexual and clonal reproduction for stand genetic structure.

Abstract

Quaking aspen (Populus tremuloides Michx.) grows throughout the northern and central portions of North America. Reproduction occurs both sexually via seeds and clonally from root suckers. Clonal delineation using morphological/phenological traits, and more recently, highly variable nuclear microsatellites have shown considerable variation in the size of clonal assemblies, and the relative importance of sexual versus clonal reproduction across the species range. In order to provide reliable estimates of genet size (N/G; ramets per sampled genet) and genotypic diversity (G/N; genets/ramets), and to compare genetic and phenotypic clone delineation, we characterized 181 sampled stems (ramets) at seven nuclear microsatellites, and morphological and phenological traits from six clones (genet size ≥11). Genotypic diversity was moderate (G/N = 0.18) and within the range reported in other studies across North America. Multivariate statistics revealed a high correspondence between genetic and phenotypic clone delineation, both with and without predefined genetic groups (94.2 %, 81.7 %). Moderate average genet size (5.6 ramets per genet) and the occurrence of genetically distinct single-ramet genets surrounded by larger genets suggested intermediate levels of sexual reproduction contributing to the genetic structure of this stand. Significant differences among genets were found for phenological and morphological traits such as bark thickness and leaf shape. However, most clones showed no significant differences in diameter growth which was likely caused by poor drainage in this high clay soil that inhibited the expression of genetic differences in growth.

     

The propagule density of Lipomyces and other yeasts in Forest Soils

Yeasts constituted a major portion of the mycoflora in a variety of forest soils. Numbers ofLipomyces were highest in the mineral soil and very low in the L horizon or upper portion of the F horizon. It is felt that the form of nitrogen in the soil plays a major role in the development of large populations ofLipomyces in soil. Yeasts (excludingLipomyces) were most numerous in the L horizon and mineral soil and least common in the F horizon. Yeast cells acounted for a smaller percentage of the total fungal propagules on hardwood litter than on conifer litter.Paper No. 3740 of the Journal Series of the North Carolina State University Agr. Exp. Sta., Raleigh, N.C. 27607.     

P2-P3 conformationally constrained ketoamide-based inhibitors of cathepsin K

An orally bioavailable series of ketoamide-based cathepsin K inhibitors with good pharmacokinetic properties has been identified. Starting from a potent inhibitor endowed with poor drug properties, conformational constraint of the P(2)-P(3) linker and modifications to P(1') elements led to an enhancement in potency, solubility, clearance, and bioavailability. These optimized inhibitors attenuated bone resorption in a rat TPTX hypocalcemic bone resorption model.