Protein misfolding and disease: from the test tube to the organism

Protein misfolding is the underlying cause of many highly debilitating disorders ranging from Alzheimer's Disease to Cystic Fibrosis. Great strides have been made recently in understanding what causes proteins to misfold, primarily through the use of biophysical and computational techniques that enable systematic and quantitative analysis of the effects of a range of different perturbations in proteins. Correlation of the results of such analyses with observations made in animal models of disease has however been limited by their seemingly irreconcilable differences in methodology and scope. Several recent studies have however begun to overcome this limitation by combining the two approaches. This strategy has made it possible to investigate many of the consequences of protein misfolding in vivo, ranging from disease pathogenesis to epigenetic regulation, in the context of the fundamental physico-chemical principles derived from extensive and highly detailed studies undertaken in vitro.     

Effects of cover reduction on mulgara Dasycercus cristicauda (Marsupialia: Dasyuridae), rodent and invertebrate populations in central Australia: Implications for land management

Abstract This study investigates the effect of cover reduction on the mulgara, Dasycercus cristicauda, a small marsupial classified as vulnerable to extinction, which occurs in areas of central Australia dominated by hummock grasslands. Loss or degradation of spinifex has been implicated in population declines of this species previously, but the importance of cover in maintaining quality habitat remains speculative. To determine the effect on D. cristicauda of cover reduction, caused by the harvesting of spinifex, we monitored population changes and changes in prey resources (rodents and invertebrates) before and after spinifex harvesting took place at a site near the Ayers Rock Resort, Northern Territory, Australia. Ten plots, each of 8.75 ha, were established and sampled from May 1994 to October 1995. Harvesting took place on five plots in August 1994, which reduced spinifex cover from 46 to 21% and the amount of spinifex >0.25 m high from 42 to 2%. Harvesting did not significantly affect the number of D. cristicauda known to be alive or captured despite other studies indicating that cover is an important habitat attribute. There was also no evidence that cover reduction impacted on the biomass of the invertebrate food resources. However, there was a significant reduction in the number of rodents captured. The lack of a response to cover reduction by D. cristicauda is possibly because the cover of Triodia remained high enough (above 15%) to sustain animals, and harvested areas were relatively small. This study therefore suggests that D. cristicauda can tolerate a moderate local reduction in cover of its preferred habitat. However, it remains possible that other land use practices that cause severe reduction of cover (including clearing for mining or fire prevention, grazing which may result in spinifex reduction through trampling, and fire management) will have more dramatic effects on D. cristicauda. Evaluation of such effects should be a priority for future research.     

Geographical gradients in seed mass in relation to climate

Aim To determine whether latitudinal and longitudinal gradients in seed mass are related to variation in climatic features including temperature, solar radiation and rainfall. Location Australia. Methods Seed mass was estimated from over 1600 provenances covering the latitudinal and longitudinal extents of 34 perennial Glycine taxa in Australia. Climatic data were obtained from ANUCLIM 5.1 for collection locations based on long‐term meteorological records across Australia. These climatic data were subject to principal components analysis to extract three components as climatic indices. Generalized linear models were used in three separate sets of analyses to evaluate whether seed mass–latitude and seed mass–longitude relationships persisted after taking climatic variation into account. First, relationships were examined across species in analyses that did not explicitly consider phylogenetic relationships. Secondly, phylogenetic regressions were performed to examine patterns of correlated evolutionary change throughout the Glycine phylogeny. Within‐species analysis was also performed to examine consistency across different taxonomic levels. Results Geographical variation in seed mass among species was related primarily to temperature and solar radiation, while rainfall was much less influential upon seed mass. Partialing out the influence of temperature and solar radiation in models resulted in the disappearance of significant seed mass–latitude and seed mass–longitude relationships. Patterns within species were generally consistent with patterns among species. However, in several species, factors additional to these climatic variables may contribute to the origin and maintenance of geographical gradients in seed mass, as significant seed mass–latitude and seed mass–longitude relationships remained after controlling for the influence of climatic variables. Main conclusions Our empirical results support the hypotheses that (1) seed mass is larger at low latitudes and in the interior of the Australian continent due to increased metabolic costs at high temperatures, and that (2) higher levels of solar radiation result in an increase in the availability of photosynthate, which in turn leads to an increase in biomass for the production of large seeds. In effect, our findings show that greater energy is available precisely where needed, that is, where high temperatures require large seed mass on the basis of metabolic requirements.     

Forest tree growth is linked to mycorrhizal fungal composition and function across Europe

Most trees form symbioses with ectomycorrhizal fungi (EMF) which influence access to growth-limiting soil resources. Mesocosm experiments repeatedly show that EMF species differentially affect plant development, yet whether these effects ripple up to influence the growth of entire forests remains unknown. Here we tested the effects of EMF composition and functional genes relative to variation in well-known drivers of tree growth by combining paired molecular EMF surveys with high-resolution forest inventory data across 15 European countries. We show that EMF composition was linked to a three-fold difference in tree growth rate even when controlling for the primary abiotic drivers of tree growth. Fast tree growth was associated with EMF communities harboring high inorganic but low organic nitrogen acquisition gene proportions and EMF which form contact versus medium-distance fringe exploration types. These findings suggest that EMF composition is a strong bio-indicator of underlying drivers of tree growth and/or that variation of forest EMF communities causes differences in tree growth. While it may be too early to assign causality or directionality, our study is one of the first to link fine-scale variation within a key component of the forest microbiome to ecosystem functioning at a continental scale.Subject terms: Microbial ecology, Forest ecology, Fungal ecology, Biogeochemistry     

Structure of the caudal neural tube in an ascidian larva: vestiges of its possible evolutionary origin from a ciliated band.

Ultrastructural analysis and differential immunocytochemical staining with two antitubulin monoclonal antibodies were used to reexamine the organization and development of the neural tube in the larva of an ascidian, Ciona intestinalis, in appraisal of a theory that the dorsal tubular nervous system of the chordates evolved from two halves of a ciliated band in an auricularia-like larva of the kind found in echinoderms and hemichordates. One of the antibodies stained cilia in the nervous system and elsewhere; the other reacted primarily with neuronal axons. The caudal neural tube consists of four rows of large ciliated ependymal-glial cells enclosing an axial neural canal into which their single cilia extend. Two ventrolateral nerve tracts, containing axons, arise in the posterior brain region and extend along the length of the caudal tube, partially surrounded by the ependymal cells. The nonnervous, ciliated, ependymal neural tube of the ascidian larva with its two associated nerve tracts survives as a primitive early condition that could result from a ciliated band transformation. Tissues in the distal-most part of the ascidian larval tail have cell lineage origins that indicate an evolutionary history different from those in the proximal majority of the tail. The ependymal cells in this presumed later addition to the tail are not ciliated, although all of the others in the caudal ependymal tube appear to be.     

Faecal indicator impacts on recreational waters: budget studies and diffuse source modelling

A series of investigations are underway which have quantified the contribution of faecal indicators delivered to nearshore coastal waters from the sewerage system and riverine inputs. Studies have been completed in Jersey, Staithes, Yorkshire, South Wales and the North-west. The research protocols have involved quantification of high and low flow faecal indicator delivery from the sewerage system and riverine sources as well as construction of nonpoint source models designed to predict faecal indicator delivery from diffuse, catchment sources. These investigations suggest a dynamic, but predictable, balance between inputs from the sewerage system and from 'catchment' sources. The sewerage system dominates during low flow conditions but is often overtaken by riverine inputs during high flow conditions after rainfall. Many bathing beach locations exhibit non-compliance after rainfall when stream inputs, rather than sewerage inputs, commonly dominate. The implications of this input pattern is that routine monitoring data may not provide information relevant to new infrastructure planning designed to achieve bathing beach compliance. This suggests that the present scientific information base is insufficient to underpin the extensive UK infra-structure investment programmes designed to ensure compliance with existing EU Directive 76/160/EEC standards. Furthermore, results to date, suggest that management attention must expand from its historical focus on infra-structure provision to incorporate diffuse sources of faecal indicator loading which present a new set of management and modelling challenges.     

Energetics of photosynthesis in Zea mays. I. Studies of the flash-induced electrochromic shift and fluorescence induction in bundle sheath cells

Bundle sheath strands free of mesophyll contamination were isolated from 3–4-week-old leaves of maize (Zea mays L.). Patterns of electron flow in the preparations were studied in the presence of physiological substrates. Relative electron flow rates were estimated from the flash-induced electrochromic band shift changes (P-518) and cytochrome f turnover. Induction of chlorophyll fluorescence was also measured. Little Photosystem II activity was found to be present, the principal pathway of electron flow being Photosystem I-driven cyclic electron transfer. The latter was activated through reductive poising by NADPH, generated via malate decarboxylation or (less efficiently) from dihydroxyacetone phosphate. The actions of these electron donors and of oxygen, nitrite and methyl viologen as electron acceptors in redox poising the Photosystem I-driven cycle were investigated and are discussed in relation to the regulation of photosynthesis in the bundle sheath.     

Sequence specificity of cytosine methylation in the DNA of the Chinese hamster ovary (CHO-K1) cell line

We have determined the DNA renaturation kinetics for those DNA sequences of the Chinese hamster ovary (CHO-K1) cells in which enzymatic cytosine methylation occurred immediately after strand synthesis and for those in which methylation was delayed after strand synthesis. DNA sequences showing immediate or delayed methylation were found to be distributed throughout all repetition classes of the DNA of these cells, with a slight concentration of immediate methylation in moderately repetitive sequences and with delayed methylation being slightly over-represented in the highly repetitive fraction. However, DNA sequences showing both classes of methylation were represented equally in unique DNA sequences. We interpret these data to mean that the methylase acting near the replication forks (the ‘immediate’ methylase) is a relatively inefficient enzyme, missing some 20% of hemimethylated sites produced by DNA replication in these cells. We suggest that the methylase performing maintenance methylation at sites remote from the replication forks (the ‘delayed’ methylase) is simply a back-up enzyme for the first and that it has no true sequence specificity. The implications of this for the function(s) of DNA methylation in mammalian cells are discussed.     

Intensive investigation in management of Hodgkin's disease.

Ninety-eight patients with clinically localised Hodgkin''s disease underwent laparotomy and splenectomy to determine the extent of microscopic spread. In 68 patients the procedure was carried out for untreated disease apparently confined above the diaphragm. Abdominal disease cannot be confidently excluded on the basis of non-invasive investigation at presentation. Clinical assessment of splenic disease was unreliable unless gross splenomegaly was present. Pedal lymphography was accurate in assessing para-aortic and iliac disease but of no value in assessing other intra-abdominal lymph node involvement, including that of the mesenteric lymph node. Trephine bone marrow biopsy findings were normal in all patients before surgery, and only one patient was found to have diseased bone marrow by Stryker-saw biopsy at operation. Liver disease was identified at operation in nine patients, some of whom were asymptomatic with clinically undetectable splenic and nodal disease. Detailed clinical staging failed to detect disease in one-third of patients who underwent laparotomy. These studies show that if radiotherapy is to remain the treatment of choice for disease truly localised to lymph nodes a detailed staging procedure, including laparotomy and splenectomy, remains essential. The value of this potentially curative treatment is considerably diminished in the patient who has been inadequately staged.