Function of calmodulin in postsynaptic densities. II. Presence of a calmodulin- activatable protein kinase activity

Because the calmodulin in postsynaptic densities (PSDs) activates a cyclic nucleotide phosphodiesterase, we decided to explore the possibility that the PSD also contains a calmodulin-activatable protein kinase activity. As seen by autoradiographic analysis of coomassie blue-stained SDS polyacrylamide gels, many proteins in a native PSD preparation were phosphorylated in the presence of [γ-(32)P]ATP and Mg(2+) alone. Addition of Ca(2+) alone to the native PSD preparation had little or no effect on phosphorylation. However, upon addition of exogenous calmodulin there was a general increase in background phosphorylation with a statistically significant increase in the phosphorylation of two protein regions: 51,000 and 62,000 M(r). Similar results were also obtained in sonicated or freeze thawed native PSD preparations by addition of Ca(2+) alone without exogenous calmodulin, indicating that the calmodulin in the PSD can activate the kinase present under certain conditions. The calmodulin dependency of the reaction was further strengthened by the observed inhibition of the calmodulin-activatable phosphorylation, but not of the Mg(2+)-dependent activity, by the Ca(2+) chelator, EGTA, which also removes the calmodulin from the structure (26), and by the binding to calmodulin of the antipsychotic drug chlorpromazine in the presence of Ca(2+). In addition, when a calmodulin-deficient PSD preparation was prepared (26), sonicated, and incubated with [γ-(32)P]ATP, Mg(2+) and Ca(2+), one could not induce a Ca(2+)-stimulation of protein kinase activity unless exogenous calmodulin was added back to the system, indicating a reconstitution of calmodulin into the PSD. We have also attempted to identify the two major phosphorylated proteins. Based on SDS polyacrylamide gel electrophoresis, it appears that the major 51,000 M(r) PSD protein is the one that is phosphorylated and not the 51,000 M(r) component of brain intermediate filaments, which is a known PSD contaminant. In addition, papain digestion of the 51,000 M(r) protein revealed multiple phosphorylation sites different from those phosphorylated by the Mg(2+)-dependent kinase(s). Finally, although the calmodulin-activatable protein kinase may phosphorylate proteins I(a) and I(b), the cyclic AMP-dependent protein kinase, which definitely does phosphorylate protein I(a) and I(b) and is present in the PSD, does not phosphorylate the 51,000 and 62,000 M(r) proteins, because specific inhibition of this kinase has no effect on the levels of the phosphorylation of these latter two proteins.     

The Emerging Role of Copeptin

Direct measurement of the nonapeptide vasopressin has been limited by analyte instability ex vivo and in vivo rapid degradation, low serum concentrations requiring a sensitive assay and inherent secretory pulsatility. Copeptin is a 39 amino acid glycopeptide cleavage product of vasopressin synthesis with high stability, providing a marker of vasopressin secretion. Copeptin measurement has applications in diagnosis of diabetes insipidus and other diseases with altered vasopressin secretion. This review summarises our current understanding of serum copeptin measurement in diabetes insipidus and possible future applications of copeptin assays. As vasopressin is a stress hormone, there is emerging evidence on the use of copeptin for diagnosis and prognostication of disorders such as syndrome of inappropriate anti-diuretic hormone secretion, diabetes mellitus, critical illness, stroke, cardiovascular disease, respiratory disease, renal disease and thermal stress. Copeptin concentration measurement is likely to improve the diagnostic reliability of diabetes insipidus and, as a marker of stress, may have diagnostic or prognostic utility in specific clinical circumstances. Further studies are needed to determine if goal-directed therapy using plasma copeptin concentrations may improve patient outcomes.     

Made in America: Immigrant Students in Our Public Schools

Made in America: Immigrant Students in Our Public Schools. Laurie Olsen New York: The New Press, 1997. 276 pp.     

Evolution at the tip and base of the X chromosome in an African population of Drosophila melanogaster

Hitchhiking effects of advantageous mutations have been invoked to explain reduced polymorphism in regions of low crossing-over in Drosophila. Besides reducing DNA heterozygosity, hitchhiking effects should produce strong linkage disequilibrium and a frequency spectrum skewed toward an excess of rare polymorphisms (compared to the neutral expectation). We measured DNA polymorphism in a Zimbabwe population of D. melanogaster at three loci, yellow, achaete, and suppressor of forked, located in regions of reduced crossing-over. Similar to previously published surveys of these genomic regions in other populations, we observed low levels of nucleotide variability. However, the frequency spectrum was compatible with a neutral model, and there was abundant evidence for recombination in the history of the yellow and ac genes. Thus, some aspects of the data cannot be accounted for by a simple hitchhiking model. An alternative hypothesis, background selection, might be compatible with the observed patterns of linkage disequilibrium and the frequency spectrum. However, this model cannot account for the observed reduction in nucleotide heterozygosity. Thus, there is currently no satisfactory theoretical model for the data from the tip and base of the X chromosome in D. melanogaster.      

Short chain oligogalacturonides induce ethylene production and expression of the gene encoding aminocyclopropane 1-carboxylic acid oxidase in tomato plants

Oligogalacturonic acids (OGAs), derived from plant cell wall pectin, have been implicated in a number of signal transduction pathways involved in growth, development and defense responses of higher plants. This study investigates the size range of OGAs capable of inducing ethylene synthesis in tomato plants, and demonstrates that in contrast with many other effects, only short chain OGAs are active. Oligomers across a range of DP from 2-15 were separated and purified to homogeneity by QAE-Sephadex anion exchange chromatography using a novel elution system. The OGAs were applied to tomato plants and assayed for their ability to induce ethylene gas release and changes in steady state levels of mRNA encoding the ethylene forming enzyme aminocyclopropane-1-carboxylic acid oxidase (ACO). The study demonstrated that only OGAs in the size range of DP4-6 were active both in eliciting ACO expression and in the production of ethylene.      

Neogene planktonic foraminiferal biostratigraphy and evolution: Equatorial to subantarctic, South Pacific

Detailed planktonic foraminiferal zonations have been established for the Neogene (Latest Oligocene through present) in six DSDP sites in the South Pacific ranging from equatorial to subantarctic latitudes (48°S). Two basic zonal schemes are readily recognized: tropical and temperate. The tropical zonation is best developed in DSDP Site 289 and the temperate zonation in Sites 206, 207A and 284. Tropical and temperate zonations can be linked by a warm subtropical scheme in Site 208, because this sequence includes a mixture of tropical and temperate elements. A site located close to the Subantarctic Convergence (Site 281) contains a zonation largely of temperate character, but the present of cooler elements and some differences in biostratigraphic ranges have required a slightly different biostratigraphic scheme.Although two broad schemes are recognized, none of the biostratigraphic sequences are identical between any of the sites. This reflects differences in biogeography, evolution and diachronous extinction at various latitudes during the entire Neogene. Diachronism in biostratigraphic ranges continue to create difficulties in correlation across such wide latitudes.Our detailed work has required the establishment of new biostratigraphic zonations in certain parts of the Neogene sequence and modifications in some other parts. Otherwise, previously established schemes are followed as closely as possible. In the temperate region, a new zonation has been established for the Early Miocene to early Middle Miocene. For the remainder of the Neogene the zonation of Kennett (1973) has been largely used. The tropical zonation of Blow (1969) is employed in the equatorial Site 289, but with further subdivisions for Zones N4 and N17. For areas intermediate between tropical and temperate latitudes (Site 208), a modified Early Miocene zonation is established based on changes in tropical and temperate elements.The zonal schemes are established on taxa that exhibit both diachronous and isochronous ranges across the latitudes. Zones that are at least partly diachronous include the Globigerinoides trilobus and Globorotalia miozea Zones of Early Miocene age; perhaps the Globorotalia mayeri Zone (its base) of the Middle Miocene; the Globorotalia conomiozea Zone of the Late Miocene; and the Globorotalia crassaformis Zone of the Early Pliocene.A large number of datum levels are recognized based on first evolutionary appearances or extinctions. The most widely applicable datums are as follows: latest Oligocene — Globigerinoides F.A.; Early Miocene — Globoquadrina dehiscens, F.A., Globorotalia kugleri L.A., Catapsydrax dissimilis L.A. and Praeorbulina glomerosa F.A.; Middle Miocene — Orbulina suturalis F.A., Globorotalia peripheroacuta F.A., Fohsella lineage L.A., Globorotalia mayeri L.A.; Late Miocene — “Neogloboquadrina” continuosa L.A., Globoquadrina dehiscens L.A., Globorotalia cibaoensis F.A.; Early Pliocene — Globorotalia puncticulata F.A., Globorotalia margaritae F.A.; Early Pleistocene — Globorotalia truncatulinoides F.A. A number of other datums are identified which assist with correlation over more restricted latitudinal ranges.The evolution of most Neogene planktonic foraminifera is now well established for a wide range of water masses. Evolutionary lineages are primarily centered in the temperate and tropical regions. Tropical lineages have recently been reviewed by Srinivasan and Kennett (1981) and are not discussed in detail here. However, Sphaeroidinellopsis seminulina is now considered to have evolved directly into S. paenedehiscens during the Late Miocene and S. subdehiscens Blow is considered to be junior synonym of S. seminulina.A new evolutionary lineage is recognized in the warm subtropics (Site 208) whereby Globigerina woodi woodi gave rise to Globigerinoides subquadratus via Globigerina brazieri. The discovery of this lineage clearly demonstrates that Globigerinoides is a polyphyletic “genus”. Another major phylogenetic lineage is recognized within the temperate globorotaliids of Early Miocene age as follows: “N.” continuosa → Globorotalia zealandica incognita → G. zealandica zelandica → G. praescitula → G. miozea. Although parts of this lineage have been recognized earlier, the entire phylogeny has previously been underscribed.A new Early to Middle Miocene lineage is recognized in the subantarctic to temperate areas which involve a transition from Globorotalia praescitula to G. challengeri n. sp. via intermediate forms.Two major Neogene globorotaliid lineages — the Menardella of the tropics and Middle Miocene to Recent forms of Globoconella of the temperate areas — are both considered to have evolved from Globorotalia praescitula beginning in the Early Miocene. This evolution initially was restricted to temperate areas but has since separated into distinctly tropical and temperate phylogenetic elements.     

The complex mechanism of antibody-mediated clearance of Bordetella from the lungs requires TLR4

Although the antibacterial effects of Abs are well studied in in vitro systems, the in vivo effects of Abs cannot always be accurately predicted. Complicated cross-talk between different effector functions of Abs and various arms of the immune system can affect their activities in vivo. Using the mouse respiratory pathogen Bordetella bronchiseptica, we examined the mechanisms of Ab-mediated clearance of bacteria from the respiratory tract. Interestingly, although TLR4 was not necessary for protective immunity following infection, it was required for rapid bacterial clearance in mice that were vaccinated or adoptively transferred Abs. TLR4 was important for the rapid recruitment of neutrophils that are necessary for Ab-mediated bacterial clearance via a mechanism that requires both FcgammaR and CR3. These data are consistent with a model in which TLR4-mediated inflammatory responses aid in the recruitment of neutrophils, which phagocytose Ab- and complement-opsonized bacteria via FcgammaRs and CR3. Although pattern recognition receptors are known to be involved in innate immunity and the generation of adaptive immunity, their contributions to specific adaptive immune functions should be considered in ongoing efforts to improve vaccine-induced protective immunity.