Human mannosidosis--the enzyme defect

Normal human liver α-mannosidase exists in at least 3 forms, separable by DEAE cellulose chromatography. The A and B forms are most active at pH 4.4 while activity of form C is maximal at pH 6.0. In two cases of mannosidosis, examined by ion exchange chromatography and isoelectric focusing, both A and B forms were absent and the residual α-mannosidase activity was due to the presence of the C form in normal amounts.     

The uptake of swainsonine, a specific inhibitor of alpha-D-mannosidase, into normal human fibroblasts in culture

Swainsonine, an indolizidine alkaloid, found in plants of the genus Swainsona, has been shown to be a strong inhibitor in vitro of the alpha-D-mannosidase activity in normal human fibroblasts. Therefore, inhibition of alpha-D-mannosidase activity in extracts of harvested cells grown with swainsonine in the medium has been used to follow the association of the alkaloid with normal human fibroblasts in culture. Swainsonine that could not be removed by extensive washing became associated with the cells within 1 min, and it is concluded that the alkaloid is internalized rapidly by the cells. The amount of swainsonine taken up into the cells depended on the length of time in contact and the concentration of swainsonine in the medium, but at 37 degrees C a plateau of internalized swainsonine occurred after 2 hr with extracellular concentrations of swainsonine of 100 microM or greater. At lower concentrations of swainsonine the rate of uptake was found to be temperature-dependent, increasing greatly at 20 degrees C. The rapidity and temperature sensitivity of the uptake, together with the observation that mannose or mannose-6-phosphate did not prevent the association, suggest that swainsonine enters the cells by permeation rather than by endocytosis. When swainsonine is withdrawn from the culture medium, there is a decrease with time of cell-associated swainsonine. The kinetics of uptake and release of swainsonine and its slightly basic nature make it likely that swainsonine is concentrated initially in the lysosomes. This rapid, but reversible, concentration of swainsonine in lysosomes would be consistent with the observed effects of the toxin in vivo.     

Spatial and environmental factors contributing to patterns in arboreal and terrestrial oribatid mite diversity across spatial scales

Recruitment is a crucial event in the plant life cycle that is very sensitive to interaction with established vegetation. Based on a large comparative experiment, we tested the hypothesis that the components of recruitment––emergence time and rate, seedling survival and biomass––differ in response to plant–plant interactions during recruitment. The consequences for the population are predicted with a simple demographic model assessing the response of seed production. In a common garden experiment, we recorded the recruitment of four target species in an individual-based survey protocol. A total of 7,680 seeds were sown within 20 neighbourhoods, consisting of 19 mono-specific herbaceous stands and a control treatment without vegetation. We measured transmitted light, temperature and moisture at soil surface to characterise the environmental conditions within neighbourhoods. The mean height of neighbours controlled temperature buffering and light interception and thus depicted the interaction gradient. Emergence rate and time increased with neighbour height in two of the four target species, while seedling survival and biomass significantly decreased with neighbour height in three and all four target species, respectively. We recorded a shift in seedling neighbour interactions under the tallest neighbours that largely favoured emergence but strongly depressed seedling survival and biomass. The components of recruitment were predicted to differ in their impact on later adult performance. Biomass strongly contributed to predicted seed production in three target species, and emergence had an equal or greater impact on a fourth species. These results confirm the fundamental role of plant–plant interactions in the recruitment of herbaceous species through a complex combination of habitat amelioration, which facilitates emergence and light competition, which in turn limits seedling survival and biomass.     

Sectoral Aggregation Error in the Accounting of Energy and Emissions Embodied in Trade and Consumption

Correctly accounting for the energy and emissions embodied in consumption and trade is essential to effective climate policy design. Robust methods are needed for both policy making and research—for example, the assignment of border carbon adjustments (BCAs) and greenhouse gas emission reduction responsibilities rely on the consistency and accuracy of such estimates. This analysis investigates the potential magnitude and consequences of the error present in estimates of energy and emissions embodied in trade and consumption. To quantify the error of embodied emissions accounting, we compare the results from the disaggregated Global Trade Analysis Project (GTAP 8) data set, which contains 57 sectors to results from different levels of aggregation of this data set (3, 7, 16, and 26 sectors), using 5,000 randomly generated sectoral aggregation schemes as well as aggregations generated using several commonly applied decisions rules. We find that some commonly applied decision rules for sectoral aggregation can produce a large error. We further show that an aggregation scheme that clusters sectors according to their energy, emissions, and trade intensities (net exports over output) can minimize error in embodied energy and emissions accounting at different levels of aggregation. This sectoral aggregation scheme can be readily used in any input‐output analysis and provide useful information for computable general equilibrium modeling exercises in which sector aggregation is necessary, although our findings suggest that, when possible, the most disaggregated data available should be used.     

Comparison of the N-acetylhexosaminidase components in the ram testis and epididymis by isoelectric focusing

1. Several enzymic components, with both N-acetyl-beta-glucosaminidase and N-acetyl-beta-galactosaminidase activity, have been demonstrated in the ram testis and epididymis by isoelectric focusing. 2. The component (I), which predominates in the testis and caput of the epididymis, is isoelectric at pH6.2+/-0.1, whereas the predominant component (II) in the epididymal isthmus and cauda is isoelectric at pH7.0+/-0.1. 3. The total activity and the relative proportions of the enzymic components vary in the different sections of the epididymis. 4. Although their pH optima are slightly different, the appropriate K(m) values for components I and II for the hydrolysis of 4-methylumbelliferyl-N-acetyl-beta-glucosaminide and N-acetyl-beta-galactosaminide and the ratios of the maximal velocities towards the two substrates are very similar.     

Aspects of vertebral development in lizards and snakes

The embryonic and postnatal development of vertebrae of the mid-trunk and mid-tail in Lacerta vivipara, Anguis fragilis and Natrix natrix have been studied. Centra and neural arches develop independently after the perichordal tube has been formed, the neurocentral suture representing a consistent boundary between them. The condyle and cup of each centrum are formed by differential growth of cartilage; in Anguis and Natrix the joints between centra are synovial. The zygosphenial joints of Lacerta are continuous with the zygapophysial joints and not separate from them as in snakes. The autotomy splits in the tail vertebrae of Lacerta are derived from four distinct components which become continuous in postnatal life. The chordal cartilage is not involved in the split formation.     

Isolation of mannose-binding proteins from human and rat liver

The interaction of e-aq., CO2-. and one-electron reduced nitroaromatics (RNO2-.) with ascorbate oxidase (AAO) was studied in aqueous solution at pH 6.0 and 7.5 by using the technique of pulse radiolysis. From observations at 330, 410 and 610 nm, interaction of e-aq. and CO2-. with AAO results in non-specific reduction of the protein followed by reduction of Type 1 Cu in a rate-determining intramolecular step. Only a few per cent of the reducing equivalents ultimately results in reduction of Type 1 Cu. With large excesses of reducing equivalents (e-aq. and CO2-.) with respect to the copper concentration, the amount of Type 1 copper reduced never exceeds 50% of the total amount of Type 1 copper after a single radiation pulse. With less-powerful reducing agents, e.g. RNO2-. reduction of Type 1 Cu occurs via a bimolecular step, and there is no evidence for formation of radicals on protein residues. From observations at 330 nm it is evident that Type 2 and/or Type 3 Cu may also be reduced along with Type 1 Cu. Almost stoichiometric reduction of AAO by RNO2-. was observed, e.g. the protein accepts 6-7 reducing equivalents. It is inferred that the various types of redox couples Cu2+/Cu+ are in equilibrium and that intramolecular electron transfer between the different types of Cu is not rate-determining when using RNO2-. as reducing agent.