Mechanism of ubiquitin activation revealed by the structure of a bacterial MoeB-MoaD complex.

The activation of ubiquitin and related protein modifiers is catalysed by members of the E1 enzyme family that use ATP for the covalent self-attachment of the modifiers to a conserved cysteine. The Escherichia coli proteins MoeB and MoaD are involved in molybdenum cofactor (Moco) biosynthesis, an evolutionarily conserved pathway. The MoeB- and E1-catalysed reactions are mechanistically similar, and despite a lack of sequence similarity, MoaD and ubiquitin display the same fold including a conserved carboxy-terminal Gly-Gly motif. Similar to the E1 enzymes, MoeB activates the C terminus of MoaD to form an acyl-adenylate. Subsequently, a sulphurtransferase converts the MoaD acyl-adenylate to a thiocarboxylate that acts as the sulphur donor during Moco biosynthesis. These findings suggest that ubiquitin and E1 are derived from two ancestral genes closely related to moaD and moeB. Here we present the crystal structures of the MoeB-MoaD complex in its apo, ATP-bound, and MoaD-adenylate forms, and highlight the functional similarities between the MoeB- and E1-substrate complexes. These structures provide a molecular framework for understanding the activation of ubiquitin, Rub, SUMO and the sulphur incorporation step during Moco and thiamine biosynthesis.     

Identification of a bacterial inhibitor against g-type lysozyme

Lysozymes are antibacterial effectors of the innate immune system in animals that hydrolyze peptidoglycan. Bacteria have evolved protective mechanisms that contribute to lysozyme tolerance such as the production of lysozyme inhibitors, but only inhibitors of chicken (c-) and invertebrate (i-) type lysozyme have been identified. We here report the discovery of a novel Escherichia coli inhibitor specific for goose (g-) type lysozymes, which we designate PliG (periplasmic lysozyme inhibitor of g-type lysozyme). Although it does not inhibit c- or i-type lysozymes, PliG shares a structural sequence motif with the previously described PliI and MliC/PliC lysozyme inhibitor families, suggesting a common ancestry and mode of action. Deletion of pliG increased the sensitivity of E. coli to g-type lysozyme. The existence of inhibitors against all major types of animal lysozyme and their contribution to lysozyme tolerance suggest that lysozyme inhibitors may play a role in bacterial interactions with animal hosts.     

Impact of bolus volume on small intestinal intra-luminal impedance in healthy subjects

AIM: To assess the impact of bolus volume on the characteristics of small intestinal (SI) impedance signals.METHODS: Concurrent SI manometry-impedance measurements were performed on 12 healthy volunteers to assess the pattern of proximal jejunal fluid bolus movement over a 14 cm-segment.Each subject was given 34 boluses of normal saline (volume from 1 to 30 mL) via the feeding tube placed immediately above the proximal margin of the studied segment.A bolus-induced impedance event occurred if there was > 12%...     

Habitat use of leatherside chub ( Gila copei ) in the presence of predatory brown trout ( Salmo trutta )

The objectives of this study were to (1) examine distribution and habitat use of leatherside chub ( Gila copei ) in a creek with high densities of predatory brown trout ( Salmo trutta ), and (2) compare study results with published accounts of leatherside chub habitat use in streams with few or no brown trout. A 14-km section of Diamond Fork Creek, Utah, was point-shocked to determine macrohabitat (main channel and lateral habitat) and microhabitat (current, depth, substrate, and cover) use of leatherside chug. At the macrohabitat level, leatherside chub were most common in lower reaches of Diamond Fork Creek where the channel was braided and lateral habitats (cutoff pools and backwaters) were abundant. Only a single leatherside chub was found in the main channel of Diamond Fork. Investigations in other systems lacking brown trout indicated that leatherside chub occupy main channel habitats. At the microhabitat level, we found significant positive associations between presence of leatherside chub and both habitat depth and silt. In addition, we found a significant negative association between presence of leatherside chub and number of brown trout. Our results suggest predation by brown trout may limit leatherside chub to lateral habitats that could provide refugia against predation. The introduction of brown trout to systems with little lateral habitat could have serious effects on persistence of resident leatherside chub populations.     

Vascular flora of Kane Lake Cirque, Pioneer Mountains, Idaho

Kane Lake Cirque lies in the western Pioneer Mountains of south central Idaho. An inventory of the high elevation flora of the cirque revealed the presence of 180 vascular taxa representing 95 genera and 30 families. Five alpine taxa are here documented from Idaho for the first time: Carex incurviformis Mack., Draba fladnizensis Wilfen., Potentilla nivea L., Ranunculus gelidus Kar. & Kir., and Ranunculus pygmaeus Wahlenb. Kane Lake Cirque also contains populations of four additional alpine taxa considered to be of conservation concern in Idaho: Erigeron humilis Graham, Parnassia kotzebuei Cham., Saxifraga adscendens L., and Saxifraga cernua L.     

Soil properties associated with vegetation patches in a Pinus ponderosa —bunchgrass mosaic

Since Euro-American settlement, fire exclusion and other factors have dramatically altered interior western coniferous forests. Once open and parklike, present-day structure in many southwestern Pinus ponderosa forests consists of dense stands of young, small-diameter trees, with small patches of larger, old trees, and relict open bunchgrass areas. Our objectives were to assess differences in soil properties associated with these different vegetation patches. We examined soil morphological characteristics, pH, organic C concentration, total N concentration, C:N ratio, and phytolith concentration from profiles within 6 transects (18 soil pedons) crossing patches of dense stands of small diameter trees, patches of old-growth trees, and open grassy areas. Results indicate that old-growth plots had significantly lower A horizon pH and thicker O horizons than grass plots. In general, we found vegetation patches had statistically similar C and N concentrations and C:N ratios for A and B horizons; however, C in the A horizon was positively correlated with O horizon accumulation ( r 2 = 0.79). Greater accumulation of organic C in the A horizon of forested areas contrasts with commonly reported results from mesic, mid-continental prairie-forest ecosystems but is typical for many arid, semiarid, and humid savanna ecosystems. Phytolith concentration was similar among old-growth pine, dense younger pine, and open grassy plots; the lack of a spatial pattern in phytolith distribution could indicate that grass cover was more spatially continuous in the past. Additionally, this interpretation is consistent with current theories regarding historical vegetation change in these forests.     

Role of serum components in the binding and phagocytosis of oxidatively damaged erythrocytes by autologous mouse macrophages

To investigate the role of autologous serum components in the recognition of damaged cells by macrophages, we examined the binding and phagocytosis of damage oxidatively damaged red blood cells with Cu2+ and ascorbate (oxRBCs) by autologous resident mouse peritoneal macrophages. The binding of oxRBCs by macrophages was independent of the presence of serum. However, phagocytosis by macrophages increased with serum concentration, and macrophages showed little ingestion of oxRBCs in a serum-free medium. Macrophages neither bound nor appreciably ingested native RBCs (before oxidation) in either the absence or presence of autologous serum. Mouse macrophages ingested significantly more native as well as oxRBCs in the presence of heat-inactivated fetal calf serum than in the presence of heat-inactivated mouse serum. Pretreated oxRBCs with normal serum were rarely ingested by macrophages in a serum-free medium. Phagocytosis of oxRBCs was significantly inhibited by depletion of IgG or calcium from serum, by heat inactivation of complement, or by antiserum against mouse C3. These results demonstrate that serum components such as IgG, C3, and calcium are involved in phagocytosis of oxRBCs by autologous macrophages.     

An Adaptive Design for Six Sigma（ ADFSS ） ： a Simulated Annealing and Regression Analysis Embedded Approach

Design for six sigma (DFSS) is a powerful approach of designing products,processes,and services with the objective of meeting the needs of customers in a cost-effective manner.DFSS activities are class...