INCOMPLETE ABSCISSION OF NEEDLE CLUSTERS AND RESIN RELEASE FROM ARTIFICIALLY WATER-STRESSED LOBLOLLY PINE (PINUS TAEDA): A COMPONENT FOR PLANT-ANIMAL INTERACTIONS

A selected Pinus taeda L. tree was rapidly and permanently water-stressed by severing and sealing the bole and observed daily for seven wk and then weekly for six months. Insects were captured before and during treatment from the stressed and adjacent control trees. Tissue samples were extracted at selected intervals for comparison of the stressed and control trees. Drying of the crown of the stressed tree commenced immediately. By the tenth day the previous yrs' needle clusters were red and dropping from the tree while the current yr's needle clusters remained green and did not fall. By the eleventh day beads of resin, predominantly α- and β-pinene, formed and dripped from the abscission scars of the youngest needle clusters of the previous yr. At this time and for several days thereafter, insects were attracted to the stressed tree. Microscopy of abscission sites of the stressed and control trees shows that the sites where resin flowed had incomplete abscission layers, lysigenous collapse of adjacent cortical cells to form a cavity that coalesced the resin ducts associated with the needle cluster, and had mechanically ruptured the tissue connections so that resin normally contained within the duct system was released to the exterior of the tree. Volatilized α-pinene is attractive to the insects associated with pines. Failure of abscission in the stressed tree leading to abnormal resin release is thus proposed as a means by which insects are attracted to stressed trees.     

Priming interactions between platelet activating factor and histamine in the in vivo microcirculation.

To elucidate whether priming exists between platelet-activating factor (PAF) and histamine in the microcirculation, we measured the clearance of FITC-dextran 150 in response to the topical applications of substimulatory concentrations of PAF and histamine. Maximal priming by PAF was observed when a 5-min interval separated the applications of 10(-9) M PAF and 10(-6) M histamine. The mean (+/- SEM) clearance resulting from this sequence of agonist administration was 7529 +/- 659 nl.2 h-1.g-1, representing a 4.5-fold enhancement in FITC-dextran 150 clearance compared with that evoked by 10(-6) M histamine alone (1664 +/- 397 nl.2 h-1.g-1). Lowering the PAF priming dose to 10(-11) M, or reversing the order of agonist addition to the microcirculation, resulted in diminished but significant responses of 3545 +/- 1143 and 4467 +/- 1170 nl.2 hr-1.g-1, respectively. Coapplication of PAF and histamine or increasing the time interval between the agonists to 15 min greatly reduced the responses to 1906 +/- 678 and 2770 +/- 837, respectively. The PAF receptor antagonist WEB 2086 (2 mg/kg i.v.), the H1 blocker pyrilamine (10 mg/kg i.v.), and leukocyte depletion with cyclophosphamide (150 mg/kg i.p.) completely abolished the PAF priming effect. In addition, the 5-lipoxygenase inhibitor RG 5901 (1 or 10 mg/kg i.v.) produced a two-thirds attenuation in PAF priming. We conclude that 1) PAF has the ability to prime the in vivo microvascular actions of histamine in both a concentration and time-dependent fashion; 2) this primed response is receptor mediated; and 3) histamine can prime the microcirculation for enhanced responses to PAF. Our data also demonstrate that leukocytes and the release of leukotrienes participate in PAF priming.     

Existence of cytosolic phospholipase D. Identification and comparison with membrane-bound enzyme.

In a wide variety of cells, phosphatidylcholine hydrolysis in response to diverse agents is catalyzed by phospholipase D (PLD) activities that are believed to be membrane-bound. Indeed, PLD has been detected in membrane fractions of several tissues and cells. We now demonstrate in various bovine tissue including lung, brain, spleen, heart, kidney, thymus, and liver as well as rat lung that a great majority of the detectable PLD activity is cytosolic. This cytosolic PLD activity differs from a less abundant membrane-bound isozyme by chromatographic mobilities on anion exchange and gel filtration columns, by substrate specificity, by substrate concentration dependence, and by divalent cation and detergent effects. Fractionation of the cytosol by anion exchange chromatography enhances PLD activity up to 20-fold, suggesting the presence in the cytosol of PLD inhibitory factor(s). We conclude that mammalian PLD exists in multiple forms and that appropriate selection of assay conditions is critical for observing PLD activity in the cytosol.     

Production of exotoxins by Aeromonas spp. at 5°C

The ability of 60 strains of Aeromonas to produce enterotoxin and haemolysin after cultivation at 5°C for 7–10 d was investigated. The strains were isolated from lamb meat, offal, carcasses and faeces, and had previously been tested for their ability to produce these exotoxins at 37°C. The results showed that some strains of Aeromonas hydrophila and A. sobria were capable of producing enterotoxin and haemolysin at 5°C, but none of the A. caviae strains tested produced these two factors. Of the 30 A. hydrophila strains investigated 25 and 27 were enterotoxigenic and haemolytic respectively. Likewise, of the 24 A. sobria strains investigated 16 and 18 were enterotoxigenic and haemolytic respectively. The results indicate that certain strains of Aeromonas species, in particular A. hydrophila and A. sobria , are of potential public health significance in meats stored at refrigeration temperature.     

An IgA monoclonal antibody directed against type III antigen on group B streptococci acts as an opsonin

We have investigated the mechanisms by which a murine IgA mAb directed against the type III Ag (IgA anti-III mAb) of group B streptococci (GBS) protects neonatal rats from lethal infection with these organisms. Purified IgA anti-III mAb enhanced phagocytosis of type III GBS by rat peritoneal macrophages in vitro by fourfold compared with phagocytosis of buffer-treated GBS. In the absence of antibody, neonatal rat serum did not promote phagocytosis, but addition of neonatal rat serum to GBS opsonized with IgA anti-III led to a sevenfold increase in phagocytosis. Heat inactivation of C destroyed the ability of neonatal rat serum to enhance phagocytosis in the presence of IgA. C3 deposition was observed when GBS coated with IgA anti-III mAb were incubated in untreated neonatal rat serum or in serum treated with Mg/EGTA. This latter observation suggested that C3 deposition occurred through activation of the alternative pathway. The control IgA mAb MOPC 315 did not enhance GBS ingestion or C3 deposition on GBS. Depletion of C in vivo by using cobra venom factor abolished the protective effect of IgA anti-III mAb in the neonatal rat model. These data suggest that the ability of this IgA to activate C further enhances its opsonic activity and may be essential for its protective effect in vivo.     

Antigen presentation by an immature myeloid dendritic cell line does not cause CTL deletion in vivo, but generates CD8+ central memory-like T cells that can be rescued for full effector function

Immature dendritic cells (DC), in contrast to their mature counterparts, are incapable of mobilizing a CD8+ CTL response, and, instead, have been reported to induce CTL tolerance. We directly addressed the impact of immature vs mature DC on CTL responses by infusing adenovirus peptide-loaded DC (of the D1 cell line) into mice that had received adenovirus-specific naive TCR-transgenic CD8+ T cells. Whereas i.v. injection of mature DC triggered vigorous CTL expansion, immature DC elicited little proliferation involving only a minority of the TCR-transgenic CTL. Even though the latter CTL developed effector functions, including cytolytic activity and proinflammatory cytokine secretion, these cells differed significantly from CTL primed by mature DC in that they did not exhibit down-regulation of CD62L and CCR7, receptors involved in trapping of T cells in the lymphoid organs. Interestingly, adoptive transfer of CTL effector cells harvested after priming by either mature or immature DC into naive recipient mice, followed by exposure to adenovirus, yielded quantitatively and qualitatively indistinguishable CTL memory responses. Therefore, in vivo priming of naive CD8+ T cells by immature DC, although failing to induce a full-blown, systemic CTL response, resulted in the formation of central memory-like T cells that were able to expand and produce IFN-gamma upon secondary antigenic stimulation.     

The cytoplasmic concentration of free calcium in platelets is controlled by stimulators of cyclic AMP production (PGD2, PGE1, forskolin)

Maximal stimulation of platelets with thrombin results in a rapid increase in cytoplasmic Ca2+ (from 0.1 microM to 1-3 microM), as measured with the fluorescent intracellular Ca2+ indicator Quin-2. Prior addition of the adenylate cyclase stimulators PGD2, PGE1 or forskolin inhibited the rise in cytoplasmic Ca2+. When added after the maximal response to thrombin was attained adenylate cyclase stimulators caused a rapid fall of cytoplasmic Ca2+ back to the original "resting" level. This effect coincides with the reversal of thrombin-induced, Ca2+-dependent protein phosphorylation, and cytoskeleton assembly. It is suggested that cAMP-dependent reactions maintain low levels of cytoplasmic Ca2+ by promoting transport and/or binding of Ca2+.