Family physician registration at locally produced short courses.

Physicians who do not take advantage of short courses that are offered within their community may miss the opportunity to learn about new services and consultants as well as to validate new information or practices with trusted colleagues. The registration patterns at short courses of all 505 family physicians in Calgary were assessed to determine whether the sociodemographic characteristics of attendees differed from those of nonattendees and whether the sociodemographic data were predictive of attendance or nonattendance. Four variables were predictive of attendance or nonattendance: certification status with the College of Family Physicians of Canada (CFPC), year of graduation, sex and hospital affiliation. Physicians who had attended four or more courses were more likely to be graduates of Canadian or US schools, to have graduated after 1969, and to be women, certificants of the CFPC and affiliated with a hospital. Universities, hospitals and professional associations planning continuing education must determine if the needs of nonattending physicians are being met through other means or if new strategies are required to ensure that these physicians have opportunities for upgrading their knowledge. Practitioners who frequently attend short courses can help planners ensure that programming is relevant.     

Tissue-specific expression and endogenous subcellular distribution of the inositol 1,3,4,5-tetrakisphosphate-binding proteins GAP1(IP4BP) and GAP1(m)

GAP1(IP4BP) and GAP1(m) belong to the GAP1 family of Ras GTPase-activating proteins that are candidate InsP4 receptors. Here we show they are ubiquitously expressed in human tissues and are likely to have tissue-specific splice variants. Analysis by subcellular fractionation of RBL-2H3 rat basophilic leukemia cells confirms that endogenous GAP1(IP4BP) is primarily localised to the plasma membrane, whereas GAP1(m) appears localised to the cytoplasm (cytosol and internal membranes) but not the plasma membrane. Subcellular fractionation did not indicate a specific co-localisation between membrane-bound GAP1(m) and several Ca2+ store markers, consistent with the lack of co-localisation between GAP1(m) and SERCA1 upon co-expression in COS-7 cells. This difference suggests that GAP1(m) does not reside at a site where it could regulate the ability of InsP4 to release intracellular Ca2+. As GAP1(m) is primarily localised to the cytosol of unstimulated cells it may be spatially regulated in order to interact with Ras at the plasma membrane.     

AGE and SEX COMPOSITION OF THE STRIPED DOLPHIN DIE-OFF IN THE WESTERN MEDITERRANEAN

During the morbillivirus epizootic that affected the Mediterranean striped dolphins ( Stenella coeruleoalba )in 1990–1992, several thous and individuals are believed to have died. In order to determine the segments of the population that were affected by the event, the sex- and age-frequency distributions of the animals that were washed ashore on the Spanish coasts were studied. The age of the animals was determined by counting dentine growth layer groups in the teeth. The results obtained indicate that both sexes were affected similarly by the disease and that the population component that suffered the largest mortality was that of sexually mature individuals, although substantial mortality of calves, probably still dependent on their mothers, was also observed. Juveniles, in contrast, appeared to be only slightly affected by the epizootic. This pattern of mortality is not consistent with the epidemiology of morbillivirus infections previously observed in other mammals. Possible explanations of this particular age distribution include behavioral factors, increased susceptibility to the disease in adults due to the effects of pollution, high temperatures and decreased food availability, or allopatric geographical distribution of population components and, therefore, of their associated mortality.     

GTP cyclohydrolase II structure and mechanism

GTP cyclohydrolase II converts GTP to 2,5-diamino-6-beta-ribosyl-4(3H)-pyrimidinone 5'-phosphate, formate and pyrophosphate, the first step in riboflavin biosynthesis. The essential role of riboflavin in metabolism and the absence of GTP cyclohydrolase II in higher eukaryotes makes it a potential novel selective antimicrobial drug target. GTP cyclohydrolase II catalyzes a distinctive overall reaction from GTP cyclohydrolase I; the latter converts GTP to dihydroneopterin triphosphate, utilized in folate and tetrahydrobiopterin biosynthesis. The structure of GTP cyclohydrolase II determined at 1.54-A resolution reveals both a different protein fold to GTP cyclohydrolase I and distinctive molecular recognition determinants for GTP; although in both enzymes there is a bound catalytic zinc. The GTP cyclohydrolase II.GMPCPP complex structure shows Arg(128) interacting with the alpha-phosphonate, and thus in the case of GTP, Arg(128) is positioned to act as the nucleophile for pyrophosphate release and formation of the proposed covalent guanylyl-GTP cyclohydrolase II intermediate. Tyr(105) is identified as playing a key role in GTP ring opening; it is hydrogen-bonded to the zinc-activated water molecule, the latter being positioned for nucleophilic attack on the guanine C-8 atom. Although GTP cyclohydrolase I and GTP cyclohydrolase II both use a zinc ion for the GTP ring opening and formate release, different residues are utilized in each case to catalyze this reaction step.     

CAPRI and RASAL impose different modes of information processing on Ras due to contrasting temporal filtering of Ca2+

The versatility of Ca2+ as a second messenger lies in the complex manner in which Ca2+ signals are generated. How information contained within the Ca2+ code is interpreted underlies cell function. Recently, we identified CAPRI and RASAL as related Ca2+-triggered Ras GTPase-activating proteins. RASAL tracks agonist-stimulated Ca2+ oscillations by repetitively associating with the plasma membrane, yet CAPRI displays a long-lasting Ca2+-triggered translocation that is refractory to cytosolic Ca2+ oscillations. CAPRI behavior is Ca2+- and C2 domain-dependent but sustained recruitment is predominantly Ca2+ independent, necessitating integration of Ca2+ by the C2 domains with agonist-evoked plasma membrane interaction sites for the pleckstrin homology domain. Using an assay to monitor Ras activity in real time, we correlate the spatial and temporal translocation of CAPRI with the deactivation of H-Ras. CAPRI seems to low-pass filter the Ca2+ signal, converting different intensities of stimulation into different durations of Ras activity in contrast to the preservation of Ca2+ frequency information by RASAL, suggesting sophisticated modes of Ca2+-regulated Ras deactivation.     

High-resolution structures reveal details of domain closure and "half-of-sites-reactivity" in Escherichia coli aspartate beta-semialdehyde dehydrogenase

Two high-resolution structures have been determined for Eschericia coli aspartate beta-semialdehyde dehydrogenase (ecASADH), an enzyme of the aspartate biosynthetic pathway, which is a potential target for novel antimicrobial drugs. Both ASADH structures were of the open form and were refined to 1.95 A and 1.6 A resolution, allowing a more detailed comparison with the closed form of the enzyme than previously possible. A more complex scheme for domain closure is apparent with the subunit being split into two further sub-domains with relative motions about three hinge axes. Analysis of hinge data and torsion-angle difference plots is combined to allow the proposal of a detailed structural mechanism for ecASADH domain closure. Additionally, asymmetric distortions of individual subunits are identified, which form the basis for the previously reported "half-of-the-sites reactivity" (HOSR). A putative explanation of this arrangement is also presented, suggesting the HOSR system may provide a means for ecASADH to offset the energy required to remobilise flexible loops at the end of the reaction cycle, and hence avoid falling into an energy minimum.     

Application of a novel method for age estimation of a baleen whale and a porpoise

Eyeballs from 121 fin whales (Balaenoptera physalus) and 83 harbor porpoises (Phocoena phocoena) were used for age estimation using the aspartic acid racemization (AAR) technique. The racemization rate (k_Asp) for fin whales was established from 15 fetuses (age 0) and 15 adult whales where age was estimated by reading growth layer groups (GLGs) in the earplugs. The (k_Asp) for harbor porpoises was derived from 15 porpoises (two calves and 13 > 1 yr old) age‐estimated by counting GLGs in the teeth and two calves classified to age based on length. The (k_Asp) values were estimated by regression of GLGs against D/L ratios. For the fin whales an (k_Asp) of 1.15 × 10^?3/yr (SE ± 0.00005) and a D/L ratio at birth [(D/L)₀] of 0.028 (SE ± 0.0012) were estimated, which is in agreement with rates for other mysticeti. For the harbor porpoises a (k_Asp) of 3.10 × 10^?3/yr (SE ± 0.0004) and a (D/L)₀ value of 0.023 (SE ± 0.0018) were estimated, which is considerably higher than found for other cetaceans. Correlation between chosen age estimates from AAR and GLG counts indicated that AAR might be an alternative method for estimating age in marine mammals.