An improved method for preparation, stabilization, and storage of house fly (Diptera: Muscidae) microsomes

An improved method for preparation and storage of insect microsomes from house fly, Musca domestica L., abdomens was developed. Microsomes were prepared in phosphate buffer fortified with glycerol, dithiothreitol, ethylenediaminetetraacetic acid, phenylmethylsulfonyl fluoride, and 1-phenyl-2-thiourea. No cytochrome P-420 was observed when abdomens were isolated by our method. No measurable loss of cytochrome P-450, cytochrome b5 or NADPH-cytochrome c (P-450) reductase levels, or methoxyresorufin O-demethylation, 7-ethoxycoumarin O-deethylation, or aryl hydrocarbon hydroxylation activities occurred when a diluted suspension (protein concentration of 2 mg/ml) of microsomes was stored at -80 degrees C for at least 2 mo.     

Ecophysiological aspects of rapid population growth in a novel migratory blackcap (Sylvia atricapilla) population: an experimental approach

Summary Blackcaps (Sylvia atricapilla) that breed in central Europe have usually migrated to Mediterranean or African wintering grounds. In the past several decades, a portion of this breeding population has started migrating to the British Isles to overwinter and this population has increased dramatically. Several factors, including higher annual survivorship (due to supplemental feeding and reduced migratiry distance), assortative mating, and enhanced reproductive success may be involved in this rapid population growth. As part of an intensive, long-term study of this population, we tested the hypothesis that the differences in photoperiod experienced by British-wintering versus Mediterranean-wintering blackcaps might lead to relatively early vernal (i.e., migratory and/or reproductive) physiological condition in members of the former group. We found that birds exposed to photoperiodic conditions that simulated migration to Britain to overwinter generally initiated vernal migratory activity earlier than birds held under conditions simulating migration to traditional wintering areas in central Spain. This difference, coupled with the shorter migratory distance to the British Isles, leads to significantly earlier estimated arrival dates for blackcaps that winter in Britain compared to central Spain. Bimodality in arrival times suggests that assortative mating on central European breeding grounds might occur between members of the different wintering populations. Males exposed to British-winter photoperiods showed significantly earlier testicular development than males kept under Spanish-winter photoperiods. Early arrival on the breeding grounds, coupled with accelerated reproductive condition, should lead to a relatively early reproductive effort, perhaps increasing average reproductive success. In general, these results support the hypothesis that differences in photoperiod on the wintering grounds may play an important role in the dynamic state of this population.     

Thievery,home ranges,and nestmate recognition inEctatomma ruidum

Summary Thievery of food items among colonies of a ponerine ant,Ectatomma ruidum was common; nonnestmates in colonies or near the colony entrances receive incoming food items and carry them to their own colony. In our study area 7 of 10 colonies were victimized by thief ants. Colonies have discrete home ranges and home range size is correlated with the number of workers in the colony. Worker ants discriminate nestmates from non-nestmates when non-nestmates are presented at colony entrances, but individuals from different colonies were not observed to engage in agonistic interactions away from nest entrances. Non-nestmates gain entrance to colonies when the entrance is unguarded. Many instances of non-nestmates being removed from colonies by residents were observed. The costs and benefits of theft under these circumstances are considered.     

Plant hormone response mutants

A variety of plant hormone response mutants has now been described, and is surveyed in this article. In addition to hormone-insensitive mutant phenotypes with defects in hormone-related features, there exist mutants apparently constitutive for the gibberellin responses, and also a mutant hyper-responsive to gibberellin. Although there is still little biochemical evidence on the nature of these mutants, the emerging picture of their genetic dominance relationships has given rise to tentative suggestions of the involvement of represser functions in hormonal control systems.     

Short-term changes in carbon-isotope discrimination identify transitions between C3 and C4 carboxylation during Crassulacean acid metabolism

Short-term measurements of instantaneous carbon-isotope discrimination have been determined from mass-spectrometric analyses of CO₂ collected online during gas exchange for the epiphytic bromeliad Tillandsia utriculata L. Using this technique, the isotopic signature of CO₂ exchange for each phase of Crassulacean acid metabolism (CAM) has been characterised. During night-time fixation of CO₂ (Phase I), discrimination () ranged from 4.4 to 6.6, equivalent to an effective carbon-isotope ratio (¹³C) of –12.3 to –14.5 versus Pee Dee Belemnite (PDB). These values reflected the gross photosynthetic balance between net CO₂ uptake and refixation of respiratory CO₂, characteristic of CAM in the Bromeliaceae. When for the relative proportion of external (p _a ) and internal (p _i) CO₂ is taken into account, calculated p _i/p _a decreased during the later part of the dark period from 0.68 to 0.48. Measurements of during Phase II, early in the light period, showed the transition between C₄ and C₃ pathways, with carboxylation being increasingly dominated by ribulose bisphosphate carboxylase (Rubisco) as increased from 10.5 to 21.2 During decarboxylation in the light period (Phase III), CO₂ leaked out of the leaf and the inherent discrimination of Rubisco was expressed. The value of calculated from on-line measurements (64.4) showed that the CO₂ lost was considerably enriched in ¹³C, and this was confirmed by direct analysis of the CO₂ diffusing out into a CO₂-free atmosphere ( ¹³C = + 51.6 versus PDB). Instantaneous discrimination was characteristic of the C₃ pathway during Phase IV (late in the light period), but a reduction in showed an increasing contribution from phosphoenolpyruvate carboxylase. The results from this non-invasive technique confirm the observations that double carboxylation involving both phosphoenolpyruvate carboxylase and Rubisco occurs during the transient phases of CAM (II and IV) in the light period.Abbreviations and Symbols CAM Crassulacean acid metabolism - H⁺ (dawn-dusk) variation in titratable acidity - ¹³C carbonisotope ratio of plant organic material, relative to Pee Dee Belemnite (vs. PDB) - discrimination against ¹³CO₂, - p _i, p _a internal, external partial pressures of CO₂ - Rubisco ribulose1,5-bisphosphate carboxylase - PAR photosynthetically active radiation - PEPCase phosphoenolpyruvate carboxylase We are grateful for financial support in respect of research grants (GR3/5360, GR3/6419) and a studentship awarded by the Natural Environment Research Council, UK.     

Transgenetic studies implicate interactions between homologous PrP isoforms in scrapie prion replication

Transgenic (Tg) mice expressing both Syrian hamster (Ha) and mouse (Mo) prion protein (PrP) genes were used to probe the mechanism of scrapie prion replication. Four Tg lines expressing HaPrP exhibited distinct incubation times ranging from 48 to 277 days, which correlated inversely with HaPrP mRNA and HaPrPC. Bioassays of Tg brain extracts showed that the prion inoculum dictates which prions are synthesized de novo. Tg mice inoculated with Ha prions had approximately 10(9) ID50 units of Ha prions per gram of brain and less than 10 units of Mo prions. Conversely, Tg mice inoculated with Mo prions synthesized Mo prions but not Ha prions. Similarly, Tg mice inoculated with Ha prions exhibited neuropathologic changes characteristic of hamsters with scrapie, while Mo prions produced changes similar to those in non-Tg mice. Our results argue that species specificity of scrapie prions resides in the PrP sequence and prion synthesis is initiated by a species-specific interaction between PrPSc in the inoculum and homologous PrPC.     

Catabolic plasmids of environmental and ecological significance

The environmental and ecological significance of catabolic plasmids and their host strains are discussed in the context of their potential application for environmental biotechnology. Included is a comprehensive list of naturally occurring discrete catabolic plasmids isolated from either natural habitats or selective enrichment studies. General properties, such as plasmid maintenance, stability and transfer, are discussed together with the techniques for plasmid detection and monitoring in the environment. The issues concerning the construction of catabolic strains with new or broader substrate ranges and the uses of monocultures or consortia for in situ treatment are addressed.     

Evaluation of the obstetric significance of some pelvic characters in an 18th century British sample of known parity status.

The excavations at Christ Church, Spitalfields (1984-86) produced a sample of 968 human skeletons which were interred between 1729 and 1859. Of these, 387 were recovered in association with coffin plates stating name, age at death, and date of death. There are 138 adult females in the named sample and the obstetric histories of 94 have been reconstructed from historical documentation. Such variables as birth spacing, number of children, and age at first and last births are known for the majority of this sample. Any individual about whose history there is any doubt has been excluded from the analysis. A middle-class group, they were largely of high nutritional status and, by the standards of the day, lived in sanitary and comfortable conditions. Both males and females have a mean age at death of 56 years. The presence or absence, the typology, the severity, the width and the length of the preauricular sulcus, the presence or absence and the number of pits on the dorsal aspect of the pubic body, sulci along the anterior sacrum adjacent to the auricular facet, and the extension of the pubic tubercle were evaluated in relation to the obstetric histories of these females. Statistical analysis has demonstrated a relationship between the presence of pubic tubercle extension and parity status and between the degree of extension and the number of children borne (P < .02). Statistically there is no significant relationship (P > .05) between either the preauricular sulcus or pubic pitting and parity status. Sacral scarring is significantly associated (P < .05) with parity status, but as it was evident in only eight females it has little practical application. Sample sizes are small, and it must be considered that statistical evaluation of larger samples might detect associations between variables not demonstrated here. There is no significant relationship between any of the cortical variants under consideration and age at death.     

Palatine torus in the Greenlandic Norse.

Physical anthropologists have long been intrigued by the distinctive oral tori expressed by the medieval Norse populations of Iceland and Greenland. To assess the temporal and spatial variation of one form of oral tori, palatine torus, observations were made on all available Greenlandic Norse skeletons, as well as on samples of medieval Icelanders and Norwegians. In terms of temporal variation, 12th to 14th century (medieval) Greenlanders from the Eastern and Western settlements exhibited higher frequencies and more pronounced expressions of palatine torus compared with early 11th century Greenlanders. The early Greenlandic sample closely approximated the medieval Icelandic and Norwegian samples for total torus frequency, although the Norwegians exhibited the trait to a less pronounced degree. As degree of expression is the most distinctive aspect of torus variation among the Norse, some combination of environmental factors, including increased masticatory stress and chronic undernutrition, probably accounts for most of the difference between settlement period and medieval Greenlanders. Although palatine torus may be hereditary in part, environmental factors play a significant role in the expression of this trait.     

The quantitative analysis of endogenous folate catabolites in human urine.

In man folates are catabolized and excreted as inactive cleaved degradation products, a mixture of pteridines and p-aminobenzoylglutamate (pABGlu) or its acetamido derivative (apABGlu). The daily rate of excretion represents the inescapable use of the vitamin in metabolic activity and thus has implications for determining the recommended dietary allowance for the vitamin. Furthermore, the rate of catabolism has been suggested to rise during pregnancy and in certain disease states. A method is described for the quantitative extraction and assay of the folate catabolites pABGlu and apABGlu in human urine. Aliquots of 24-h urine collections are acidified and applied to columns of Dowex 50W cation-exchange resin. The catabolites are selectively batch-eluted with increasing concentrations of HCl. The fraction containing pABGlu is diazotized and then applied to a C18 Sep Pak column for further purification and concentration. The fraction containing apABGlu was deacetylated and reapplied to the Dowex column and then treated identically to the pABGlu fraction. The methanolic concentrates of both extracts were evaporated to dryness and reconstituted with water and pABGlu was regenerated by reductive cleavage of the diazotized material with Zn/HCl. The extracts of the two catabolites were separated by reverse-phase HPLC using a Radial Pak C18 column. Recovery of isolated material was monitored by the addition of high specific activity tritiated labels of both compounds added as internal standards to all urine aliquots prior to purification and analysis.