Specific changes in the in vitro production of prostanoids by the ovine cervix at parturition

A method of tissue superfusion has been used to measure in vitro prostanoid production by the ovine cervix during late pregnancy and at parturition. In late pregnancy (105-135 days of gestation) cervical tissue produced relatively large amounts of prostaglandin E (PGE); in comparison, the production rates of prostaglandin F (PGF), 13, 14-dihydro-15-oxo-prostaglandin F (PGFM) and 6-oxo-prostaglandin F1 alpha were generally low. Thromboxane B2 (TXB2) production was minimal and often unmeasurable. There were significant increases in the production rates of PGE and 6-oxo-PGF1 alpha by cervical tissue taken immediately after delivery, when compared to late pregnancy. Mean production rates of PGE increased from 19.8 +/- 4.1 to 43.8 +/- 7.4 ng/g. dry wt./min; 6-oxo-PGF1 alpha production rates increased more than three-fold from 10.0 +/- 2.7 to 34.6 +/- 9.8 ng/g. dry wt./min (means +/- S.E.M.). There were no significant differences in the rates of production of PGF, PGFM and TXB2 by the two groups.     

Injection locking and mode selection in TEA-CO2laser oscillators

The injection of a master oscillator signal in a high-power TEA-CO2laser is analyzed and a dynamic model is formulated to represent the interaction. Based on the competition between the injected signal and the spontaneous emission, the model describes the transient evolution of the different field amplitudes and phases together with their effect on the inversion. A study over a wide range of injection levels and detuning frequencies clearly indicates three distinct regions of operation: a spontaneous oscillation zone, a mode-selection zone, and a frequency-locking zone. The main predictions of the model are compared with the results obtained with an experimental injection apparatus that assures adequate control of the TEA laser cavity length and that provides means for measuring the frequency of the output pulse. While the first two zones are directly observed, it is experimentally established that, at injection levels up to 5 W/cm², the frequency-locking zone does not exceed, as predicted, the 3-MHz resolution limit of the apparatus.     

Effects of 2,2'-dipyridyl and related compounds on platelet prostaglandin synthesis and platelet function

2,2'-dipyridyl, a chelator of ferrous iron and inhibitor of platelet aggregation, was studied together with several similar compounds to determine the mechanism of their effects on platelets. All of these compounds were more potent inhibitors of arachidonic-acid-mediated aggregation (IC50, 0.17-1.8 mM) than of ADP-mediated aggregation (IC50, 7.6-19.7 mM). At low concentrations required to inhibit arachidonic-acid-mediated aggregation, 2,2'-dipyridyl, 4,4'-dipyridyl and 2-chloropyridine specifically inhibited the platelet cyclo-oxygenase. The mechanism of inhibition of ADP-induced aggregation was investigated, but was not explained. At concentrations needed to inhibit ADP-induced aggregation, 2,2'-dipyridyl did not alter cell ultrastructure, serotonin or nucleotide content or interfere with release of [14C]arachidonic acid or calcium movements. Therefore, our results indicate that 2,2'-dipyridyl and related compounds have two effects on platelets, both due to the unprotonated form. The inhibition of cyclo-oxygenase by low concentrations of these compounds is not due to bidentate iron chelation, since 4,4'-dipyridyl was almost as effective as 2,2'-dipyridyl, but is compatible with binding of these inhibitors to the iron in the heme of the cyclo-oxygenase.     

Thermal and structural behavior of milk fat. 1. Unstable species of anhydrous milk fat

The thermal and structural behavior of anhydrous milk fat (AMF) was studied by a technique that allowed simultaneous time-resolved synchrotron X-ray diffraction as a function of temperature (XRDT) and high sensitivity differential scanning calorimetry (DSC) to be carried out in the same apparatus from the same sample. In this paper (the first of a series), the less stable crystalline structures made by triacylglycerols (TG) of bulk AMF after its liquid quenching down to -8 degrees C are addressed The coexistence of two lamellar structures characterized by sharp long spacing reflections corresponding to well-defined 3L (70 A) and 2L (47 A) longitudinal stackings but broad short spacing lines related to poorly ordered hexagonal (alpha) lateral packing is shown for the first time, The bilayered structure was very unstable, since it disappeared during a 20-min isothermal recording. Simultaneous DSC and X-ray monitoring of AMF heating in the range -8, +50 degrees C at a rate of 2 degrees C/min allows the same sample to be followed on the evolution of these unstable forms to more stable varieties. The 3L stacking transforms into a new 2L crystalline structure characterized by broad LS reflections corresponding to a ill-defined 2L (37 A) longitudinal stacking but a more compact orthorhombic (beta') lateral packing. A delimitation of the domains of existence of the crystalline structures resulted from the comparison of detailed analysis of the evolutions of positions, intensities, and widths of X-ray peaks as a function of temperature to microcalorimetry recording.     

Reduced growth factor requirements and accelerated cell-cycle kinetics in adult human melanocytes transformed with SV40 large T antigen

Melanomas develop with high frequency in transgenic mice in which oncogenic sequences of the SV40 DNA tumor virus have been specifically targeted to melanocytes. To investigate the role of SV40 in melanomagenesis, cultured human melanocytes were transformed with a retroviral shuttle vector encoding the SV40 large T antigen and examined for changes in cell-cycle kinetics and growth-factor dependence. Colonies expressing the viral oncogene were morphologically indistinguishable from their non-T-antigen-transformed counterparts. Also like normal melanocytes, the infected cells remained anchorage dependent and non-tumorigenic in nude mice. However, T-antigen-positive cultures exhibited significantly accelerated population doubling times, increased saturation densities with highly confluent monolayers and a three- to fourfold extended life span. Most interestingly, cell-cycle analysis revealed a measurable shift from quiescent to cycling cells in T-antigen-expressing cultures and an acquired ability to progress more rapidly through G1. Moreover, T-antigen-positive melanocytes proliferated in the absence of PMA and required markedly reduced levels of exogenous bFGF. These studies indicate that the viral oncogen of simian virus 40 provides melanocytes with distinct growth advantages that may render these cells unusually susceptible to additional environmental challenges necessary for full expression of the malignant phenotype.     

Residues Val254, His256, and Phe259 of the angiotensin II AT1 receptor are not involved in ligand binding but participate in signal transduction

The role of the external third of helix VI of the angiotensin II (AII) AT1 receptor for the interaction with its ligand and for the subsequent signal transduction was investigated by individually replacing residues 252-256 by Ala, and residues 259 or 261 by Tyr, and permanently transfecting the resulting mutants to Chinese hamster ovary (CHO) cells. Binding experiments showed no great changes in affinity of any of the mutants for AII, [Sar1]-AII, or [Sar1, Leu8]-AII, but the affinity for the nonpeptide antagonist DuP753 was significantly decreased. The inositol phosphate response to AII was remarkably decreased in mutants V254A, H256A, and F259Y. These results indicate that AT1 residues Val254, His256, and Phe259 are not involved in ligand binding but participate in signal transduction. Based in these results and in others from the literature, it is suggested that, in addition to the His256 imidazole ring, the Phe259 aromatic ring interacts with the AII's Phe8, thus contributing to the signal-triggering mechanism.