Sub-Inhibitory Cefsulodin Sensitization of E. coli to β-lactams Is Mediated by PBP1b Inhibition

The combination of antibiotics is one of the strategies to combat drug-resistant bacteria, though only a handful of such combinations are in use, such as the β-lactam combinations. In the present study, the efficacy of a specific sub-inhibitory concentration of cefsulodin with other β-lactams was evaluated against a range of Gram-negative clinical isolates. This approach increased the sensitivity of the isolates, regardless of the β-lactamase production. The preferred target and mechanism of action of cefsulodin were identified in laboratory strains of Escherichia coli, by examining the effects of deleting the penicillin-binding protein (PBP) 1a and 1b encoding genes individually. Deletion of PBP1b was involved in sensitizing the bacteria to β-lactam agents, irrespective of its O-antigen status. Moreover, the use of a sub-inhibitory concentration of cefsulodin in combination with a β-lactam exerted an effect similar to that one obtained for PBP1b gene deletion. We conclude that the identified β-lactam/cefsulodin combination works by inhibiting PBP1b (at least partially) despite the involvement of β-lactamases, and therefore could be extended to a broad range of Gram-negative pathogens.     

Molecular motors: strategies to get along

The majority of active transport in the cell is driven by three classes of molecular motors: the kinesin and dynein families that move toward the plus-end and minus-end of microtubules, respectively, and the unconventional myosin motors that move along actin filaments. Each class of motor has different properties, but in the cell they often function together. In this review we summarize what is known about their single-molecule properties and the possibilities for regulation of such properties. In view of new results on cytoplasmic dynein, we attempt to rationalize how these different classes of motors might work together as part of the intracellular transport machinery. We propose that kinesin and myosin are robust and highly efficient transporters, but with somewhat limited room for regulation of function. Because cytoplasmic dynein is less efficient and robust, to achieve function comparable to the other motors it requires a number of accessory proteins as well as multiple dyneins functioning together. This necessity for additional factors, as well as dynein's inherent complexity, in principle allows for greatly increased control of function by taking the factors away either singly or in combination. Thus, dynein's contribution relative to the other motors can be dynamically tuned, allowing the motors to function together differently in a variety of situations.     

Sparging and agitation-induced injury of cultured animals cells: Do cell-to-bubble interactions in the bulk liquid injure cells?

It has been established that the forces resulting from bubbles rupturing at the free air (gas)/liquid surface injure animal cells in agitated and/or sparged bioreactors. Although it has been suggested that bubble coalescence and breakup within agitated and sparged bioreactors (i.e., away from the free liquid surface) can be a source of cell injury as well, the evidence has been indirect. We have carried out experiments to examine this issue. The free air/liquid surface in a sparged and agitated bioractor was eliminated by completely filling the 2-L reactor and allowing sparged bubbles to escape through an outlet tube. Two identical bioreactors were run in parallel to make comparisons between cultures that were oxygenated via direct air sparging and the control culture in which silicone tubing was used for bubble-free oxygenation. Thus, cell damage from cell-to-bubble interactions due to processes (bubble coalescence and breakup) occurring in the bulk liquid could be isolated by eliminating damage due to bubbles rupturing at the free air/liquid surface of the bioreactor. We found that Chinese hamster ovary (CHO) cells grown in medium that does not contain shear-protecting additives can be agitated at rates up to 600 rpm without being damaged extensively by cell-to bubble interactions in the bulk of the bioreactor. We verified this using both batch and high-density perfusion cultures. We tested two impeller designs (pitched blade and Rushton) and found them not to affect cell damage under similar operational conditions. Sparger location (above vs. below the impeller) had no effect on cell damage at higher agitation rates but may affect the injury process at lower agitation intensities (here, below 250 rpm). In the absence of a headspace, we found less cell damage at higher agitation intensities (400 and 600 rpm), and we suggest that this nonintuitive finding derives from the important effect of bubble size and foam stability on the cell damage process. (c) 1996 John Wiley & Sons, Inc.     

Community dynamics in relation to management of heathland vegetation in Scotland

The paper describes studies of post-fire succession in heathland vegetation in N.E. Scotland, dominated by Calluna vulgaris. A preliminary model (Legg, 1978) suggested good agreement between simulation of succession on the basis of a Markov chain and observations of stands at different stages of development after burning, at least in the earlier stages. Vegetation transitions are currently being recorded in permanent plots on burnt areas. First results confirm the view that (a) the post-fire succession has the properties of a Markov process, (b) this type of model remains valid when constructed from records of actual transitions, rather than data obtained by inference from evidence of transition. Comparing successional events in stands where, at the time of burning, the Calluna population was in pioneer-, building-, mature-and degenerate phases, shows that transition matrices generally agree with the Markov hypothesis, but not in the case of stands where Calluna was degenerate when burnt. The composition of establishing vegetation 1 year after fire is not confined to species normally associated with the early stages of succession, but reflects the composition of the stand before burning. Redevelopment after fire is described in terms of an initial floristic composition of species with strategies permitting early re-establishment, selected by the recurrence of the fire factor. Subsequent transitions represent changes in their relative abundance due to differing growth properties and competitive interactions. This interpretation applies only under conditions of recurrent incidence of fire (normally once in 10–15 yr). If fire does not recur, Calluna stands pass into the degenerate phase, where changes in the nature of relay floristics may come into play (e.g. with tree colonization).Nomenelature follows Clapham, Tutin & Warburg (1962) for vascular plants; Smith (1978) for bryophytes.     

A conceptual framework for the spatial analysis of functional trait diversity

Functional trait diversity is a popular tool in modern ecology, mainly used to infer assembly processes and ecosystem functioning. Patterns of functional trait diversity are shaped by ecological processes such as environmental filtering, species interactions and dispersal that are inherently spatial, and different processes may operate at different spatial scales. Adding a spatial dimension to the analysis of functional trait diversity may thus increase our ability to infer community assembly processes and to predict change in assembly processes following disturbance or land‐use change. Richness, evenness and divergence of functional traits are commonly used indices of functional trait diversity that are known to respond differently to large‐scale filters related to environmental heterogeneity and dispersal and fine‐scale filters related to species interactions (competition). Recent developments in spatial statistics make it possible to separately quantify large‐scale patterns (variation in local means) and fine‐scale patterns (variation around local means) by decomposing overall spatial autocorrelation quantified by Moran's coefficient into its positive and negative components using Moran eigenvector maps (MEM). We thus propose to identify the spatial signature of multiple ecological processes that are potentially acting at different spatial scales by contrasting positive and negative components of spatial autocorrelation for each of the three indices of functional trait diversity. We illustrate this approach with a case study from riparian plant communities, where we test the effects of disturbance on spatial patterns of functional trait diversity. The fine‐scale pattern of all three indices was increased in the disturbed versus control habitat, suggesting an increase in local scale competition and an overall increase in unexplained variance in the post‐disturbance versus control community. Further research using simulation modeling should focus on establishing the proposed link between community assembly rules and spatial patterns of functional trait diversity to maximize our ability to infer multiple processes from spatial community structure.     

Artificial neural networks for qualitative and quantitative analysis of target proteins with polymerized liposome vesicles

We investigate the feasibility of using the luminescence response of polymerized liposomes incorporating ethylenediaminetetraacetate europium(III) (EDTA-Eu(3+)) for monitoring protein concentrations in aqueous media. Quantitative analysis is based on the linear relationship between the luminescence enhancement of the lanthanide ion and protein concentration. Analytical figures of merit are presented for carbonic anhydrase, human serum albumin, gamma-globulins, and thermolysin. Qualitative analysis is based on the luminescence lifetime of the liposome sensor. This parameter, which follows well-behaved single exponential decays and provides characteristic values for each of the four studied proteins, demonstrates the selective potential for protein identification. Then partial least squares-1 and artificial neural networks are compared toward the quantitative and qualitative analysis of human serum albumin and carbonic anhydrase in binary mixtures without previous separation at the concentration levels found in aqueous humor.     

Cytology and sexuality of 11 species of Elatostema (Urticaceae) in limestone karsts suggests that apomixis is a recurring phenomenon

Limestone karsts across southern China to southeastern Asia are renowned biodiversity hotspots. The karst are characterized by exposed calcareous rocks, seasonal droughts and thin soils that are deficient in N and P but with high Ca and Mg content. The stressful habitat may result in high biodiversity through mechanisms such as niche differentiation, hybridization, polyploidy and apomixis. The genus Elatostema (Urticaceae) has particularily high species diversity in this area and can be used a model genus to explore the mechanisms of speciation. We conducted cytological studies on 11 species of Elatostema from 12 populations in Guangxi, China. We found five populations to be diploid (2n = 26), and seven triploid populations (2n = 39). We infer x = 13 as the basic chromosome number of Elatostema. The chromosome numbers 2n = 26 and 2n = 39 were both found in populations of E. longistipulum, indicating that this species comprise both diploids and triploids. Both male and female plants of Elatostema were found to be diploid. In contrast, the triploids found were all female plants, and these produce seeds, presumably by apomixis. We found no clear relationship between ploidy level or reproductive pathway and endemism in Elatostema which might be because Elatostema species are wind‐pollinated and independent of biotic pollinators. However, a random sample of 11 karst species revealed that ca 2/3 appeared to be apomictic, suggesting that it is a widespread reproductive strategy of Elatostema in the limestone karsts of Guangxi. Apomixis enables plants to reproduce and disperse from a single individual, allowing ‘hopeful monsters’ adapted to a new habitat to form stable populations.