Changing patterns of vitellin-related peptides during development of the cattle tick Boophilus microplus

The major components of protein extracts from the cattle tick Boophilus microplus eggs and larvae of various ages were characterized by molecular sieving chromatography, ion exchange chromatography and SDS-PAGE. The fractions analysed showed a changing chromatographic pattern development. A serum raised against the components of a fraction showing characteristics of vitellin strongly reacted in Western blots with the major peptides of extracts from eggs, larvae, gut and ovary. Comparison of patterns obtained by electrophoresis in non-denaturing PAGE, stained with Coomassie blue or with benzidine/hydrogen peroxide, revealed that the major proteins of these extracts are haemoproteins, possibly in different aggregation states or heterogeneous in composition.     

Catabolism of aggrecan proteoglycan aggregate components in short-term explant cultures of tendon.

Hyaluronan (HA) and link protein are essential components of the aggrecan proteoglycan aggregate, whereby HA binds multiple aggrecan monomers, an interaction which is stabilised by link protein. In this study, we have examined the turnover of the aggregate components, HA, link protein, and the N-terminal G1 domain of aggrecan, in explant cultures of tissue from compressed and tensional regions of young and mature bovine tendons. Western blot analyses revealed the release of highly processed link protein and G1-containing metabolites, in the absence of catabolic agents, indicating an increased turnover of these components in tendon. In addition, significant levels of HA were released from the tissue matrix into the media compartment. Furthermore, RT-PCR analysis showed that the mRNA expression of link protein and enzymes/proteins associated with HA metabolism may be modulated in the distinct functional regions of tendon with development. Perturbation of normal aggrecan aggregate metabolism may lead to tissue dysfunction.     

Studies of the M15 β-Galactosidase Complementation Process

M15 -Galactosidase was activated by heat-denatured wild-type -galactosidase, urea, and heat-denatured wild-type -galactosidase, a peptide made up of residues 6–44 of -galactosidase and CB2, the peptide that is normally used for complementation (residues 3–92 of -galactosidase). In each case roughly equal activation levels were attained. Heat-denatured wild-type -galactosidase was present as a finely divided visible white precipitate both before and after complementation. The heat-denatured protein by itself did not migrate on native PAGE and both the protein and the activity that occurred as a result of the complementation also remained at the point of application. The N-terminal ends of the heat-denatured wild-type -galactosidase must have been available for complementation and must have been mobile enough to allow tetramer to form despite being aggregated. -Galactosidase denatured by both urea and heat resulted in a streak of interacting protein on the native PAGE. Upon activation, a streak (indicating that interaction was still occurring) was still present, but it moves more slowly. Complementation using a peptide called XP (made up of residues 6–44 plus an additional nine C-terminal amino acids) resulted in three discrete forms of active enzyme at ratios of peptide to M15 -galactosidase monomer of less than 1:1. The fastest migrating of the three bands predominated at ratios near 1:1. A single active tetrameric form of M15 -galactosidase was formed with CB2. In both of these last two cases an active slow-moving diffuse band also formed (possibly a dimer of the tetramer). A quantitation of the amount of peptide bound to M15 -galactosidase by titration with XP and with CB2 and by using gel filtration after an excess of fluorescent-labeled XP was added showed that peptide bound in a 1:1 ratio (peptide/monomer) when full activity was achieved. These fluorescent studies also showed that peptide initially bound to dimer and that the tetramer was then formed.     

Serotonin,Eating Behavior,and Fat Intake

There is an intimate relationship between nutritional intake (eating) and serotonin activity. Experimental manipulations (mainly neuropharmacological) of serotonin influence the pattern of eating behavior, subjective feelings of appetite motivation, and the response to nutritional challenges. Similarly, nutritional manipulations (food restriction, dieting, or altered nutrient supply) change the sensitivity of the serotonin network. Traditionally, serotonin has been linked to the macronutrient carbohydrate via the intermediary step of plasma amino acid ratios. However, it has also been demonstrated that 5-HT drugs will reduce energy intake and reverse body weight gain in rats exposed to weight increasing high fat diets. 5-HT drugs can also reduce food intake and block weight gain of rats on a high fat cafeteria diet. Some diet selection studies in rats indicate that the most prominent reduction of macronutrient intake is for fat. These data indicate that 5-HT activity can bring about a reduction in fat consumption. In turn, different types of dietary fat can alter brain 5-HT activity. In human studies the methodology of food choice experiments has often precluded the detection of an effect of 5-HT manipulation on fat intake. However, there is evidence that in obese and lean subjects some 5-HT drugs can readily reduce the intake of high fat foods. Data also suggest that 5-HT activation can lead to a selective avoidance of fat in the diet. These effects of 5-HT on the intake of dietary fat may involve a pre-absorptive mechanism and there is evidence that 5-HT is linked to cholecystokinin and enterostatin. These proposals have theoretical and practical implications and suggest possible strategies to intensify or advance fat-induced satiety signals.     

One-minute exposure of 4-cell mouse embryos to glucose overcomes morula block in CZB medium

One-cell mouse embryos that block at the 2-cell stage can progress to the morula stage in CZB medium, but fail to cavitate and then swell and lyse. A 1-min exposure to 27 mM glucose at the 4-cell stage (～42 hr) will support a high frequency of development to the blastocyst stage (75%) in the same medium. A glucose exposure is beneficial anytime between 30 and 54 hr of culture (67–73% blastocysts). Of a group of additional sugars and glucose analogues tested for their ability to replace glucose, only galactose was equivalent in promoting embryo development to the blastocyst stage (64% blastocysts). © 1994 Wiley-Liss, Inc.     

Behavior and biology of the tomato psyllid, Bactericerca cockerelli, in response to the Mi-1.2 gene

The Mi‐1.2 gene, identified from wild varieties of tomato, Solanum peruvianum (Mill) (Solanaceae), has been incorporated into near‐isogenic commercial varieties of tomato and has been shown to confer resistance to three different species of phloem feeders: aphids, whiteflies, and nematodes. The results presented here show that plants bearing Mi‐1.2 were also resistant to the tomato psyllid, Bactericerca [Paratrioza] cockerelli (Sulc) (Homoptera: Psyllidae), a serious pest of tomato, Solanum lycopersicon (Mill), in the western half of North America. In choice studies, tomato psyllids preferred to settle on plants that did not contain the gene [Moneymaker (mi‐1.2)] compared to near‐isogenic plants with the gene [Motelle (Mi‐1.2)]. As a result, total oviposition was higher on the susceptible variety, although no‐choice studies indicated that there were no differences in numbers of eggs laid by individual females on either variety. Survival from egg to adult was higher on plants lacking the gene compared to plants containing the gene. However, there were no differences in total development time of individuals reared from either variety. The results suggest that mechanisms of resistance to the tomato psyllid observed in plants bearing the Mi‐1.2 gene are distinct from the mechanisms of resistance to the three phloem feeders examined in other studies.     

Factors influencing the en route survivorship and post-voyage growth of a common ship biofouling organism,Bugula neritina

The likelihood that viable non-indigenous biofouling species will survive a voyage on a vessel is influenced by a range of factors, including the speed, duration, and route of the voyage and the amount of time the vessel spends in port. In this study, a land-based dynamic flow device was used to test the effect of recruit age, vessel speed and voyage duration on the survivorship and growth of the bryozoan Bugula neritina. In the experiment, one-week-old recruits had a higher likelihood (100%) of surviving voyages than older (one–month-old, 90%) or younger (one-day-old, 79%) recruits, but survival was not influenced by vessel speed (6 and 18 knots) or voyage duration (two and eight days). The results suggest that the non-indigenous species B. neritina can be effectively transferred at a range of ages but one-week-old recruits are more likely to survive the translocation process and survive in the recipient environment.