Absolute polarity determination of teeth cementum by phase sensitive second harmonic generation microscopy

The absolute sign of local polarity in relation to the biological growth direction has been investigated for teeth cementum using phase sensitive second harmonic generation microscopy (PS-SHGM) and a crystal of 2-cyclooctylamino-5-nitropyridine (COANP) as a nonlinear optic (NLO) reference material. A second harmonic generation (SHG) response was found in two directions of cementum: radial (acellular extrinsic fibers that are oriented more or less perpendicular to the root surface) and circumferential (cellular intrinsic fibers that are oriented more or less parallel to the surface). A mono-polar state was demonstrated for acellular extrinsic cementum. However, along the different parts of cementum in circumferential direction, two corresponding domains were observed featuring an opposite sign of polarity indicative for a bi-polar microscopic state of cellular intrinsic cementum. The phase information showed that the orientation of radial collagen fibrils of cementum is regularly organized with the donor (D) groups pointing to the surface. Circumferential collagen molecules feature orientational disorder and are oriented up and down in random manner showing acceptor or donor groups at the surface of cementum. Considering that the cementum continues to grow in thickness throughout life, we can conclude that the cementum is growing circumferentially in two opposite directions and radially in one direction. A Markov chain type model for polarity formation in the direction of growth predicts D-groups preferably appearing at the fiber front.

Intercellular adhesion and membrane polarity in rat hepatocytes: Localization of cell -CAM 105 and other domain-specific membrane proteins {\it in situ\/} and {\it in vitro\/} by electron microscopy

Cell-CAM 105 has been identified as a cell adhesion molecule (CAM) based on the ability of monospecific and monovalent anti-cell-CAM 105 antibodies to inhibit the reaggregation of rat hepatocytes. Although one would expect to find CAMs concentrated in the lateral membrane domain where adhesive interactions predominate, immunofluorescence analysis of rat liver frozen sections revealed that cell-CAM 105 was present exclusively in the bile canalicular (BC) domain of the hepatocyte. To more precisely define the in situ localization of cell-CAM 105, immunoperoxidase and electron microscopy were used to analyze intact and mechanically dissociated fixed liver tissue. Results indicate that although cell-CAM 105 is apparently restricted to the BC domain in situ, it can be detected in the pericanalicular region of the lateral membranes when accessibility to lateral membranes is provided by mechanical dissociation. In contrast, when hepatocytes were labeled following incubation in vitro under conditions used during adhesion assays, cell-CAM 105 had redistributed to all areas of the plasma membrane. Immunofluorescence analysis of primary hepatocyte cultures revealed that cell-CAM 105 and two other BC proteins were localized in discrete domains reminscent of BC while cell-CAM 105 was also present in regions of intercellular contact. These results indicate that the distribution of cell-CAM 105 under the experimental conditions used for cell adhesion assays differs from that in situ and raises the possibility that its adhesive function may be modulated by its cell surface distribution. The implications of these and other findings are discussed with regard to a model for BC formation.^ Analysis of molecular events involved in BC formation would be accelerated if an in vitro model system were available. Although BC formation in culture has previously been observed, repolarization of cell-CAM 105 and two other domain-specific membrane proteins was incomplete. Since DMSO had been used by Isom et al. to maintain liver-specific gene expression in vitro, the effect of this differentiation system on the polarity of these membrane proteins was examined. Based on findings presented here, DMSO apparently prolongs the expression and facilitates polarization of hepatocyte membrane proteins in vitro. ^

Molecular mechanisms involved in the regulation of cell polarity and genome stability by the p21 -activated kinase, Shk1, in the fission yeast, Schizosaccharomyces pombe

The essential p21-activated kinase (PAK), Shk1, is a critical component of a Ras/Cdc42/PAK complex required for cell viability, normal cell polarity, proper regulation of cytoskeletal dynamics, and sexual differentiation in the fission yeast, Schizosaccharomyces pombe. While cellular functions of PAKs have been described in eukaryotes from yeasts to mammals, the molecular mechanisms of PAK regulation and function are poorly understood. This study has characterized a novel Shk1 inhibitor, Skb15, and, in addition, identified the cell polarity regulator, Tea1, as a potential biological substrate of Shk1 in S. pombe. Skb15 is a highly conserved WD repeat protein that was discovered from a two-hybrid screen for proteins that interact with the catalytic domain of Shk1. Molecular data indicate that Skb15 negatively regulates Shk1 kinase activity in S. pombe cells. A null mutation in the skb15 gene is lethal and results in deregulation of actin polymerization and localization, microtubule biogenesis, and the cytokinetic machinery, as well as a substantial uncoupling of these processes from the cell cycle. Loss of Skb15 function is suppressed by partial loss of Shk1, demonstrating that negative regulation of Shk1 by Skb15 is required for proper execution of cytoskeletal remodeling and cytokinetic functions. A mouse homolog of Skb15 can substitute for its counterpart in fission yeast, demonstrating that Skb15 protein function has been substantially conserved through evolution. ^ Our laboratory has recently demonstrated that Shk1, in addition to regulating actin cytoskeletal organization, is required for proper regulation of microtubule dynamics in S. pombe cells. The Shk1 protein localizes to interphase and mitotic microtubules, the septum-forming region, and cell ends. This pattern of localization overlaps with that of the cell polarity regulator, Tea1, in S. pombe cells. The tea1 gene was identified by Paul Nurse's laboratory from a screen for genes involved in the control of cell morphogenesis in S. pombe. In contrast to wild type S. pombe cells, which are rod shaped, tea1 null cells are often bent and/or branched in shape. The Tea1 protein localizes to the cell ends, like Shk1, and the growing tips of interphase microtubules. Thus, experiments were performed to investigate whether Tea1 interacts with Shk1. The tea1 null mutation strongly suppresses the loss of function of Skb15, an essential inhibitor of Shk1 function. All defects associated with the skb15 mutation, including defects in F-actin organization, septation, spindle elongation, and chromosome segregation, are suppressed by tea1Δ, suggesting that Tea1 may function in these diverse processes. Consistent with a role for Tea1 in cytokinesis, tea1Δ cells have a modest cell separation defect that is greatly exacerbated by a shk1 mutation and, like Shk1, Tea1 localizes to the septation site. Molecular analyses showed that Tea1 phosphorylation is significantly dependent on Shk1 function in vivo and that bacterially expressed Tea1 protein is directly phosphorylated by recombinant Shk1 kinase in vitro. Taken together, these results identify Tea1 as a potential biological substrate of Shk1 in S. pombe. ^ In summary, this study provides new insights into a conserved regulatory mechanism for PAKs, and also begins to uncover the molecular mechanisms by which the Ras/Cdc42/PAK complex regulates the microtubule and actin cytoskeletons and cell growth polarization in fission yeast. ^

Structural group composition of low-polarity organic matter dissolved in waters of the Northwest Indian Ocean and the Red Sea

Data on amounts of various functional groups, i.e. aldehyde, acid, ester, alcohol, thiol and aromatic groups in several fractions of low-polarity dissolved organic matter are presented. An assumption that this organic matter is part of the lipid fraction is not confirmed. Amount of aromatic compounds in waters of the Northwest Indian Ocean is estimated to be about 1000 times higher than quantity of aromatic hydrocarbons discharged into the ocean each year in petroleum and petroleum products.

Characteristic directions and polarity ratings and polarity chron assignments of ODP Leg 171B sites

The full suite of magnetic polarity chrons from Subchron M''-2r'' (early Albian) through Chron C13r (latest Eocene) were resolved at one or more Ocean Drilling Program sites on the Blake Nose salient of the Florida continental margin. These sediments preserve diverse assemblages of calcareous and siliceous microfossils; therefore, the composite suite provides a reference section for high-resolution correlation of biostratigraphic datums to magnetic polarity chrons of the Late Cretaceous and Paleogene. Relative condensation or absence of polarity zones at different sites along the transect enhance the recognition and dating of depositional sequences and unconformities within the margin succession. A stable paleolatitude of ~25°N was maintained from the late Aptian through Eocene.

Revised geomagnetic polarity timescale for the late Oligocene to early Miocene of ODP Site 177-1090

At Ocean Drilling Program (ODP) Site 1090 (subantarctic South Atlantic), benthic foraminiferal stable isotope data (from Cibicidoides and Oridorsalis) span the late Oligocene through early Miocene (~24-16 Ma) at a temporal resolution of ~5 ky. Over the same interval, a magnetic polarity stratigraphy can be unequivocally correlated to the geomagnetic polarity time scale (GPTS), thereby providing direct correlation of the isotope record to the GPTS. In an initial age model, we use the newly derived age of the Oligocene/Miocene (O/M) boundary of 23.0 Ma of Shackleton et al. (2000, doi:10.1130/0091-7613(2000)28<447:ACAFTO>2.0.CO;2), revised to the new astronomical calculation (La2003) of Laskar et al (2004, doi:10.1016/j.icarus.2004.04.005) to recalculate the spline ages of Cande and Kent (1995, doi:10.1029/94JB03098). We then tune the Site 1090 dekta18O record to obliquity using La2003. In this manner, we are able to refine the ages of polarity chrons C7n through C5Cn.1n. The new age model is consistent, within one obliquity cycle, with previously tuned ages for polarity chrons C7n through C6Bn from Shackleton et al. (2000) when rescaled to La2003. The results from Site 1090 provide independent evidence for the revised age of the Oligocene/Miocene boundary of 23.0 Ma. For early Miocene polarity chrons C6AAr through C5Cn, our obliquity-scale age model is the first to allow a direct calibration to the GPTS. The new ages are generally within one obliquity cycle of those obtained by rescaling the Cande and Kent (1995) interpolation using the new age of the O/M boundary (23.0 Ma) and the same middle Miocene control point (14.8 Ma) used by Cande and Kent (1995).

Relationship between definitive datum levels and magnetic polarity and revised geomagnetic polarity time-scale at DSDP Hole 72-516F

This paper contains magnetobiostratigraphic correlation charts for each of the four sites occupied during DSDP Leg 72. Microfossil zonal boundaries and magnetic polarity determinations for Sites 515 through 518 are summarized in Figures 1 through 4, respectively. Our discussion focuses on the correlations derived for the Paleogene and late Cretaceous (Coniacian-Maestrichtian) of Site 516, because of the value of this site as a stratigraphic reference section for the South Atlantic.

Lower Jaramillo polarity transition records from ODP Holes 108-665A and 121-758B

Two records of the geomagnetic polarity transition at the beginning of the Jaramillo Subchron (0.97 Ma) have been obtained from sediments in the equatorial Atlantic (Leg 108, Site 665; 2.95°N, 340.33°E) and Indian (Leg 121, Site 758; 5.38°N, 90.35°E) oceans. Both cores yielded high-quality magnetostratigraphic results; however, the relatively low sedimentation rates, the weak magnetizations, and complex demagnetization behavior of some transitional samples suggest that the record of the transitional field behavior may be less reliable. In addition, variations in grain size preclude reliable paleointensity determinations although the remanence in both cores is apparently dominated by magnetite. Despite these possible complications, the two cores yield transitional paths that are neither far-sided nor near-sided. Together with published records that meet minimum reliability standards, the two equatorial records presented here suggest that the lower Jaramillo transitional field morphology was significantly nonaxisymmetric. The mean normal and reversed inclinations from both cores deviate from the inclination expected from a geocentric axial dipole, as noted in virtually all marine sediment cores. The observed inclinations provide further support for a polarity-dependent nondipole contribution to the time-averaged field.

Plio-Pleistocene magnetic polarity stratigraphies of ODP Leg 177 Sites from the South Atlantic

Magnetic polarity stratigraphies from ODP Leg 177 'high resolution' sites indicate Brunhes sedimentation rates in the 12-25 cm/kyr range, with a trend of decreasing sedimentation rates with increasing age. Magnetite is the principal remanence-carrying mineral. Downcore alteration of magnetite and authigenic growth of iron sulfides introduces a high coercivity diagenetic remanence carrier (pyrrhotite). The change in pore water sulfate with depth in the sediment tends to be in step with the decrease in magnetization intensity, indicating the link between sulfate reduction and magnetite dissolution. Shipboard pass-through magnetometer data are generally very noisy due to a combination of weak magnetization intensities, drilling-related core deformation, and the influence of authigenic iron sulfides. Post-cruise progressive demagnetization of discrete samples aids the magnetostratigraphic interpretation, as these measurements are less influenced by low magnetization intensities and drilling-related deformation. The magnetostratigraphic interpretations provide much-needed calibration for biostratigraphic events in the high latitude southern oceans. Apart from the ODP Hole 745B (Kerguelen Plateau), published Plio-Pleistocene magnetostratigraphies from ODP sites in the Southern Ocean are poorly constrained. For this reason, we compare interpolated ages of 11 radiolarian events and one diatom event that occur at Hole 745B and Leg 177 sites.