Immunoreactivity of the <Emphasis Type="Italic">Mycobacterium avium</Emphasis> subsp. <Emphasis Type="Italic">paratuberculosis</Emphasis> 19-kDa lipoprotein

Background  

The Mycobacterium tuberculosis 19-kDa lipoprotein has been reported to stimulate both T and B cell responses as well as induce a number of Th1 cytokines. In order to evaluate the Mycobacterium avium subsp. paratuberculosis (M. avium subsp. paratuberculosis) 19-kDa lipoprotein as an immunomodulator in cattle with Johne's disease, the gene encoding the 19-kDa protein (MAP0261c) was analyzed.     

Thermodynamic linkage between the S1 site,the Na+ site,and the Ca2+ site in the protease domain of human activated protein C (APC). Sodium ion in the APC crystal structure is coordinated to four carbonyl groups from two separate loops

The serine protease domain of activated protein C (APC) contains a Na+ and a Ca2+ site. However, the number and identity of the APC residues that coordinate to Na+ is not precisely known. Further, the functional link between the Na+ and the Ca2+ site is insufficiently defined, and their linkage to the substrate S1 site has not been studied. Here, we systematically investigate the functional significance of these two cation sites and their thermodynamic links to the S1 site. Kinetic data reveal that Na+ binds to the substrate-occupied APC with K(d) values of approximately 24 mm in the absence and approximately 6 mm in the presence of Ca2+. Sodium-occupied APC has approximately 100-fold increased catalytic efficiency ( approximately 4-fold decrease in K(m) and approximately 25-fold increase in k(cat)) in hydrolyzing S-2288 (H-d-Ile-Pro-Arg-p-nitroanilide) and Ca2+ further increases this k(cat) slightly ( approximately 1.2-fold). Ca2+ binds to the protease domain of APC with K(d) values of approximately 438 microm in the absence and approximately 105 microm in the presence of Na+. Ca2+ binding to the protease domain of APC does not affect K(m) but increases the k(cat) approximately 10-fold, and Na+ further increases this k(cat) approximately 3-fold and decreases the K(m) value approximately 3.7-fold. In agreement with the K(m) data, sodium-occupied APC has approximately 4-fold increased affinity in binding to p-aminobenzamidine (S1 probe). Crystallographically, the Ca2+ site in APC is similar to that in trypsin, and the Na+ site is similar to that in factor Xa but not thrombin. Collectively, the Na+ site is thermodynamically linked to the S1 site as well as to the protease domain Ca2+ site, whereas the Ca2+ site is only linked to the Na+ site. The significance of these findings is that under physiologic conditions, most of the APC will exist in Na2+-APC-Ca2+ form, which has 110-fold increased proteolytic activity.     

Longevity determines sizes of an adult intertidal barnacle

The small honeycomb barnacle Chamaesipho tasmanica Foster and Anderson often forms continuous sheets covering the substratum at mid-shore levels of sheltered rocky shores but also occurs in sparse distributions on exposed shores, and on higher levels of sheltered shores, in south-eastern Australia. Larger barnacles are generally found in more exposed and higher areas. Effects of each site on ultimate sizes of barnacles were therefore examined by measuring growth of barnacles of the same cohort at sheltered and exposed shores from the end of 1989 to the end of 1992. Because Chamaesipho varied in size among sites, and because size was not necessarily representative of age, three growth models were proposed to explain this size differential. While rates of growth and periods of growth did not differ, differences in longevity were found to influence size so that larger barnacles survived longer and were more abundant on the more exposed shores. In fact, when barnacles were aged, it was found that a greater proportion of older barnacles (>3 years of age) occupied these exposed areas. It seems that site-specific characteristics influenced longevity in some places so that larger Chamaesipho continued to predominate in these areas.     

Induced resistance to Mexican bean beetles in soybean: variation among genotypes and lack of correlation with constitutive resistance

Fourteen genotypes (varieties) of soybean (Glycine max) were screened for levels of induced resistance to Mexican bean beetle (Epilachna varivestis) damage, and a subset of 6 of those varieties was screened for levels of constitutive resistance to Mexican bean beetles. Experiments were carried out in the greenhouse, with damage imposed by Mexican bean beetle larvae, and levels of resistance measured by a choice test bioassay with adult beetles. We found significant variation among soybean genotypes in levels of both induced and constitutive resistance. We found no significant correlation between levels of induced and constitutive resistance measured in the same genotypes. We compare these results to past work on resistance in the soybean-Mexican bean beetle system, consider the implications of variation in both types of resistance for plant-herbivore interactions in agricultural and natural systems, and discuss the relationship between induced and constitutive resistance. Received: 30 November 1998 / Accepted: 25 June 1999     

Prey selection and foraging behaviour by Pterostichus cupreus L. (Col., Carabidae) under laboratory conditions

Abstract: Several authors have indicated that carabid beetles offer potential as predators of insect pests on arable crops, but this potential is reduced by their limited ability to climb the crop plants. In the current investigation an initial laboratory experiment indicated that seven species of carabid beetles commonly found in arable fields in the UK would feed on both aphid and collembolan prey. Pterostichus cupreus L. and Bembidion guttula Fabr. were found to accept both live and dead aphids and collembolans. In further laboratory investigations P. cupreus exhibited greater consumption of the cereal aphid Metopolophium dirhodum Walker than the alternative prey species Heteromurus nitidus , Templeton (Collembola, Entomobryidae) and there was some indication of preference for the aphid prey as these were often consumed first. Significantly more handling time was required for the consumption of M. dirhodum compared to H. nitidus. The treatments where P. cupreus was given a diet of aphids or collembolans prior to the prey choice experiment, did not appear to influence the number or type of prey items consumed.
Using a four-arm olfactometer P. cupreus reacted positively to the volatiles from collembolans, but gave a less clear response to those from the cereal aphid M. dirhodum . In an artificial field arena P. cupreus readily climbed barley plants in search of M. dirhodum , perhaps as a response to volatile emissions, but this behaviour changed when H. nitidus were introduced onto the substrate of the arena. Although the current results indicated that P. cupreus has potential for the control of cereal aphids, the presence of alternative prey appeared to reduce this potential and suggest that the carabid is an opportunistic feeder.     

Appropriate experimental design for transplanting mussels (Mytilus sp.) in analyses of environmental stress: an example in Sydney Harbour (Australia)

Some locations in Sydney Harbour (Sydney, Australia) contain large amounts of contaminants (heavy metals and hydrocarbons), sometimes in concentrations thought to affect biological systems. In order to estimate effects of sediment-bound contaminants on the physiology of organisms living above the sediment, the rates of clearance and respiration and the efficiency of absorption of mussels, Mytilus sp., living in a contaminated location were measured, the scope for growth (SfG) was calculated and compared to that of mussels living in uncontaminated locations. Two different models were proposed to explain expected differences. The first was that the contaminants at the impacted location reduced the SfG of local mussels; the second was that at the contaminated location only those mussels survived that had a small SfG (genetic differences between populations might be a reason for differential survival). To test which model applied, mussels were transplanted between the contaminated and uncontaminated locations. Moving and disturbing mussels (handling, tagging and caging) required the inclusion of two control treatments. These treatments were essential for a proper evaluation of the results but have generally not been included in similar studies. Effects of moving were estimated by translocating mussels from the uncontaminated and from the contaminated location to similar locations. To estimate effects of disturbance, mussels in the experimental locations were given the same treatment as the experimental mussels, but were returned to the place of origin. It was predicted that translocating and disturbing mussels would have no effect on the SfG, which would be similar to that of mussels at the place of origin. As expected, SfG was smaller (because rate of clearance was reduced) in undisturbed mussels at the contaminated location compared with those of mussels at the uncontaminated location. Because there were significant effects of disturbance on the SfG of mussels at the contaminated location, it was concluded that this difference was not caused by differences between the amounts of contaminants in the two locations, but was caused by other confounding factors (physical disturbance by crabs and fouling organisms). That the interpretation of the results would have been different if proper controls were not included is discussed in this paper and the importance of proper experimental controls is stressed.     

Effects of height on the shore and complexity of habitat on abundances of amphipods on rocky shores in New South Wales, Australia

Patterns of abundance of epifaunal crustaceans were compared between two common brown algae in intertidal rock pools on a shore near Sydney, New South Wales, Australia. Amphipods were significantly more abundant on Sargassum globulariaefolium (which was most common low on the shore) than on Hormosira banksii (more common at mid-shore levels). Experiments tested the hypotheses that height on the shore and structural complexity of the algae were the main factors influencing patterns of abundance of amphipods. Reciprocal transplants using defaunated plants and similar experiments using artificial plants allowed simultaneous tests of these hypotheses. The abundance of a common intertidal amphipod, Sunampithoe graxon was significantly smaller on Sargassum transferred from low to mid-tidal level of the shore than on any other treatments, indicating that height on the shore affects numbers of this species. There were no Sunampithoe on Hormosira even when transplanted to low shore areas, indicating that other factors, apart from the height on the shore, are also important for this amphipod. Another common amphipod, Hyale maroubrae was generally found in greatest abundances on Hormosira transplanted from mid- to low shore and on Sargassum low on the shore, again indicating the importance of height on the shore. There was no difference in abundance of amphipods on ‘complex’ and ‘simple’ artificial plants indicating that structural complexity (as defined in this study) did not influence these amphipods.