Regulation of maturation-promoting factor by protein kinase C in Chaetopterus oocytes

Summary

We present the results of a variety of studies showing that activation of protein kinase C (PKC) in oocytes of Chaetopterus pergamentaceus results in germinal vesicle breakdown (GVBD). Phorbol esters and diacylglycerol can initiate a morphologically normal GVBD accompanied by a spectrum of associated biochemical processes, including increased protein phosphorylation, a shift in protein synthesis and activation of a protein kinase, maturation promoting factor (MPF). MPF activation is essential for GVBD in response to phorbol esters. In addition, inhibitors of PKC can block naturally-induced GVBD. We also present evidence that PKC can phosphorylate p34^cde2, the catalytic subunit of MPF and that phosphorylation by PKC increases the histone H1 kinase activity of immunoprecipitated MPF. Immunoblot studies show that Chaetopterus oocyte p34^cdc2 is not tyrosine phosphorylated prior to the initiation of GVBD, indicating that activation of MPF at GVBD in this species does not require p80^cdc25, the activator of MPF at mitosis. These results suggest that PKC is an essential regulator of GVBD which can directly phosphorylate and regulate p34^cdc2. Since PKC is the intracellular receptor for and is directly activated by tumor-promoters, tumor promotion might involve acceleration of the cell cycle through modification of the enzymatic activity of MPF by PKC.     

Analysis of Tyrosinase Mutations Associated With Tyrosinase-Related Oculocutaneous Albinism (OCAI)

Mutations of the tyrosinase gene associated with a partial or complete loss of enzymatic activity are responsible for tyrosinase related oculocutaneous albinism (OCA1). A large number of mutations have been identified and their analysis has provided in-sight into the biology of tyrosinase and the pathogenesis of these different mutations. Missense mutations produce their effect on the activity of an enzyme by altering an amino acid at a specific site. The location of these mutations in the peptide can be used to indicate potential domains important for enzymatic activity. Missense mutations of the tyrosinase polypeptide cluster in four regions, suggesting that these are important functional domains. Two of the potential domains involve the copper binding sites while the others are likely involved in substrate binding. More critical analysis of the copper binding domain of tyrosinase can be gained by analyzing the structure of hemocyanin, a copper-binding protein with a high degree of homology to tyrosinase in the copper binding region. This analysis indicates a single catalytic site in tyrosinase for all enzymatic activities.     

The Living, Endogenous Stages of the Rat Coccidium, Eimeria nieschulzi

SYNOPSIS. The living, endogenous stages of Eimeria nieschulzi Dieben, 1924 (Landers isolate) were studied under the phase contrast microscope. Active sporozoites were found as early as 2.5 hours after exposure and as late as 48 hours after exposure. The first generation schizont was recognized by the presence of a refractile globule remaining from the sporozoite. First and second generation merozoites were only weakly motile and had small paired organelles. Third generation merozoites were seen 48–120 hours after exposure and were strongly motile from 72 hours after exposure onward. The paired organelle consisted of 2 intertwining portions, one 5.5 μ long, the other tapering to a slender filament and continuing to about the posterior quarter of the parasite. The fourth generation merozoites were short, curved, and weakly motile. A paired organelle about 3 μ long was seen. Gametocytes and gametes were seen 144–192 hours after exposure. Macrogametes appeared to elaborate refractile granules in the vicinity of the nucleus. No motility of any type was seen in the macrogametes. Microgametocytes were recognized when nuclear material moved to the periphery of the parasite for the formation of microgametes. Observations on living organisms agreed generally with those made on fixed and stained organisms with the exception that the living merozoites were about 20% larger.     

Gyrfalcon Falco rusticolus post-glacial colonization and extreme long-term use of nest-sites in Greenland

Gyrfalcons Falco rusticolus use the same nest-sites over long periods of time, and in the cold dry climate of Greenland, guano and other nest debris decay slowly. Nineteen guano samples and three feathers were collected from 13 Gyrfalcon nests with stratified faecal accumulation in central-west and northwest Greenland. Samples were ¹⁴C dated, with the oldest guano sample dating to c . 2740–2360 calendar years (cal yr) before present (BP) and three others were probably > 1000 cal yr BP. Feather samples ranged from 670 to 60 cal yr BP. Although the estimated age of material was correlated with sample depth, both sample depth and guano thickness gave a much less reliable prediction of sample age than use of radiocarbon dating on which the margin of error was less. Older samples were obtained from sites farther from the current Greenland Ice Sheet and at higher elevations, while younger samples were closer to the current ice sheet and at lower elevations. Values for δ¹³C showed that Gyrfalcons nesting farther from the Greenland Ice Sheet had a more marine diet, whereas those nesting closer to the ice sheet (= further inland) fed on a more terrestrial diet. The duration of nest-site use by Gyrfalcons is a probable indicator of both the time at which colonization occurred and the palaeoenvironmental conditions and patterns of glacial retreat. Nowhere before has such extreme long-term to present use of raptor nest-sites been documented.     

Effects of Temperature and Various Red or Far-red Irradiations on Phytochrome- and Gibberellin A3-mediated Germination Control in Partially Hydrated Lettuce Seeds

Dark reversion of phytochrome in partially hydrated lettuceseeds (Lactuca sativa cv. Grand Rapids) is temperature dependent.After initial red irradiation (R) the higher the storage temperature,the higher the dark reversion rate. Following dark moist storage(DMS) at 30 ?C for 15 d none of the seeds receiving initialR germinated, whereas seeds stored at 0 ?C germinated nearlyas well (about 80%) as unstored controls. The half-time fordark reversion at 20 ?C and 30 ?C is 9 d and 3 d respectively.Repeated R treatments given at 5 d intervals during DMS at 20?C and 30 ?C maintained a high germination capacity. With threeor more R treatments the effect of high temperature largelydisappeared. Dark reversion of phytochrome was not observed in partiallyhydrated lettuce seeds receiving continuous red irradiation(cont R) for two or more days. The promotive effect of contR could be reversed at any time with a brief far-red irradiation(FR), indicating that the phytochrome system remained fullyphototransformable. With continuous far-red light (cont FR)the ability of gibberellin A₃ (GA₃) to stimulate germinationdisappeared and response to GA₃ also diminished in cont R followedby FR but at a slower rate indicating the induction of secondarydormancy in these partially hydrated seeds. This induction ofdormancy was retarded by repetitive or cont R but was enhancedby cont FR. The results of this study suggest a role for theaccumulated stable intermediates of phytochrome transformationin partially hydrated seeds with repeated or continuous R treatmentsand different effects of GA₃ and R in the regulation of germination. Key words: Phytochrome, Lactuca sativa, Seed germination, Temperature, Dark reversion of phytochrome, Seed water content     

Temporal,Spatial, and Landscape Habitat Characteristics of Moose—Vehicle Collisions in Western Maine

Abstract: We analyzed moose (Alces alces)-vehicle collisions (MVCs) in western Maine, USA, from 1992 to 2005 (n = 8,156) using Geographic Information Systems to identify patterns of temporal and spatial distribution and develop predictive models based on road and landscape characteristics. We used chi-square and correlation analyses to assess temporal characteristics of MVCs, K-function and kernel analyses to identify spatial clusters of MVCs, and logistic regression to relate covariates for traffic, land-cover, land-form, and relative moose abundance to probability of MVC. We evaluated candidate models using Akaike's Information Criterion, area under the receiver operating characteristic curve (AUC), and the percentage of correctly classified observations. Most (81.6%) MVCs occurred from May to October, with peak monthly frequencies in June (18.6%). Moose-vehicle collisions were clustered spatially on roads at local (0–4 km) and regional scales (22–41 km and 45–54 km), but not at intermediate scales. Traffic-related covariates predicting MVCs included traffic volume and speed limit. For each additional 500 vehicles/day, odds of a location being an MVC increased by 57%. For each 8-km/hr increase in speed limit, odds of an MVC increased by 35%. Landscape composition covariates best predicted MVCs within a 2.5-km radius of the collision site. Mean percent cover within 2.5 km of MVCs was comprised of 36% more cutover forest, 10% more coniferous forest, 5% less deciduous-mixed forest, and 10% less nonwoody wetland than for random points. For every 5% increase in percent cutover and coniferous forest within 2.5 km of the road, predicted odds of MVC increased by 36% and 19%, respectively. Landscape configuration covariates best predicted MVCs within the 5.0-km radius. Moose-vehicle collisions were associated with areas of less interspersion of cover types; for each 5% increase in an index of interspersion-juxtaposition, predicted odds of MVC decreased by 11%. Our final model attained high predictive power (AUC = 0.835) and validation accuracy (75.0%). The model also proved robust to physiographic variation, exhibiting high predictive power (AUC = 0.828) and validation accuracy (68.8%). Managers seeking to prioritize resources for reducing MVCs or predicting future areas of high MVC probability should assess land-cover composition and configuration surrounding MVC hotspots at geographic extents out to 2.5–5 km and use this information to plan expensive roadside management practices such as fencing. The importance of traffic and landscape covariates in our modeling suggests that effective management to reduce MVCs will require a complex combination of driving speed reductions and modifications to forest management along roads.