Choline metabolism in the cerebral cortex of guinea pigs. Stable-bound acetylcholine

1. The turnover of synaptosomal (vesicular-cytoplasmic) and stable-bound (vesicular) acetylcholine isolated from cortical tissue was investigated after the administration, under local anaesthesia, of [N-Me-(3)H]choline into the lateral ventricles of guinea pigs. 2. Radioactive acetylcholine and choline present in acid extracts of subcellular fractions were separated by a combination of liquid and column ion-exchange and thin-layer chromatography. 3. The specific radioactivity and pattern of labelling of acetylcholine present in a fraction of monodisperse synaptic vesicles was found to be essentially the same as that of synaptosomal acetylcholine. 4. The specific radioactivity of stable-bound acetylcholine present in partially disrupted synaptosomes (fraction H) at short times (10-20min) after the injection of [N-Me-(3)H]choline was very variable and inversely related to the yield of acetylcholine in that fraction. 5. Evidence was found for the existence of two small, but highly labelled pools of acetylcholine, one which could be isolated in fraction H and the other which was lost when synaptosomes, after isolation by gradient centrifugation, were left at 0 degrees C or pelleted. 6. It is concluded that the results are best explained by metabolic differences among the nerve-ending compartments (thought to be vesicles) which contain stable-bound acetylcholine. Computer simulation of our experiments supports this possibility and suggests that the highly labelled pool in fraction H is present in vesicles close to the external membrane.     

Scale,succession and complexity in island biogeography: are we asking the right questions?

• 1 This paper offers a commentary on the development of island ecological theory since the publication of MacArthur & Wilson’s equilibrium theory in the 1960s. I distinguish the simple model at the core of their Equilibrium Theory of Island Biogeography (ETIB) and the broader body of their theory, which embraces evolutionary as well as ecological patterns — all, however, within the overarching framework or assumption of equilibrium.
• 2 The basic problems with the ETIB have long been known, and its status as a ruling paradigm has been the subject of concern for more than two decades. With the development of nonequilibrium ideas in ecology, island biogeographers arguably now have viable theoretical frameworks to set alongside or around the ETIB. Four conditions are highlighted as extremes: i) dynamic equilibrium; ii) dynamic nonequilibrium; iii) ‘static’ equilibrium; and iv) ‘static’ nonequilibrium: together providing a conceptual framework for island ecological analyses.
• 3 The importance of scale is stressed and attention is drawn to Haila’s spatial‐temporal continuum as an organizational device. It is argued that the processes represented within the ETIB (and by extension, other island theories) may be prominent within only a limited portion of this continuum, while elsewhere they are generally subsumed by other dominant processes.
• 4 Colonization and ecosystem development of near‐shore islands constitute just a special case of ecological succession, and thus the development of theories of island assembly may benefit accordingly from efforts to incorporate ideas from the ecological succession literature.
• 5 The desirability of specifying answerable questions is stressed, as is the need to build a greater degree of complexity into the development of island ecological models. Notwithstanding which, it is also recognized that key advances are often brought about by simple, but bold models, of the form exemplified elsewhere in this issue.

     

Design of an Insulin Analog with Enhanced Receptor Binding Selectivity: RATIONALE, STRUCTURE, AND THERAPEUTIC IMPLICATIONS*

Insulin binds with high affinity to the insulin receptor (IR) and with low affinity to the type 1 insulin-like growth factor (IGF) receptor (IGFR). Such cross-binding, which reflects homologies within the insulin-IGF signaling system, is of clinical interest in relation to the association between hyperinsulinemia and colorectal cancer. Here, we employ nonstandard mutagenesis to design an insulin analog with enhanced affinity for the IR but reduced affinity for the IGFR. Unnatural amino acids were introduced by chemical synthesis at the N- and C-capping positions of a recognition α-helix (residues A1 and A8). These sites adjoin the hormone-receptor interface as indicated by photocross-linking studies. Specificity is enhanced more than 3-fold on the following: (i) substitution of Gly^A1 by d-Ala or d-Leu, and (ii) substitution of Thr^A8 by diaminobutyric acid (Dab). The crystal structure of [d-Ala^A1,Dab^A8]insulin, as determined within a T₆ zinc hexamer to a resolution of 1.35 Å, is essentially identical to that of human insulin. The nonstandard side chains project into solvent at the edge of a conserved receptor-binding surface shared by insulin and IGF-I. Our results demonstrate that modifications at this edge discriminate between IR and IGFR. Because hyperinsulinemia is typically characterized by a 3-fold increase in integrated postprandial insulin concentrations, we envisage that such insulin analogs may facilitate studies of the initiation and progression of cancer in animal models. Future development of clinical analogs lacking significant IGFR cross-binding may enhance the safety of insulin replacement therapy in patients with type 2 diabetes mellitus at increased risk of colorectal cancer.     

How resilient are Andean montane forest bird communities to habitat degradation?

The threatened forest habitats of the tropical Andes are reportedly being modified and destroyed 30% faster than their lowland tropical counterparts, but impacts on the hyper-diverse resident avifauna have received little systematic study. We present a baseline analysis of the effects of habitat modification on birds in a lower montane forest landscape in Ecuador, comparing avian community composition in landscape elements subjected to different levels of human modification: primary forest, secondary forest, edge habitat and agricultural land. We use data from a point count survey of 300 counts at 150 sites to test whether community composition and density of birds with different reported habitat preferences and foraging strategies change among landscape elements. Species richness and diversity were lowest in agricultural land, but on some measures, equally low in primary forest. Richness and diversity peaked in secondary forest and edge habitat, but ordination and density analysis revealed clear differences in their species composition. While secondary forest contained mostly forest-preferring species, edge habitat harboured a mix of forest and open-land birds. There was a clearly structured gradient in species composition across landscape elements, with densities of habitat specialists, foraging guilds and families varying considerably from primary forest to agricultural land. Agricultural land was characterised by an assemblage of widespread, abundant species very different from that in core forest habitats. As such, while the majority of montane forest birds appear resilient to a certain level of habitat modification, they cannot persist, and are displaced, where forest has been cleared outright. We argue that, for Andean montane forests, preservation of mature secondary forest offers flexibility in supplementing preserved primary forest areas to provide sufficient habitat for the persistence of this incredibly diverse but severely threatened bird community.     

Unifying and distinguishing diversity ordering methods for comparing communities

Diversity indices have been widely used in ecological research, but they remain problematic in that different indices may rank communities inconsistently. This problem can be solved by using diversity ordering methods, the output of which is a diversity profile in graphical form for each community being compared. In this paper, we demonstrate that existing diversity ordering methods can be classified into four groups and that within-group methods are essentially equivalent, while among-group methods are not. We find that the intrinsic diversity-related methods—i.e., the group containing the right tail-sum method, the logarithmic dominance plot, the majorization method, and the k-dominance plot—provide the most stringent test of diversity ordering, and we recommend the right tail-sum method as the method of preference for practical purposes.