Role of activated macrophages in Acanthamoeba keratitis

The purpose of this study was to determine whether activating the conjunctival macrophages would affect the course of Acanthamoeba spp. keratitis in a Chinese hamster model of this disease. Chinese hamster spleen cells were stimulated with concanavalin A (Con A), and interferon gamma (IFN-gamma) -containing supernatants were collected 24 hr later. The IFN-gamma-containing supernatants were loaded into liposomes, which were fed to peritoneal macrophages in vitro. Macrophage activation was assessed by testing for production of nitric oxide (NO) with the use of Griess reagent. Conjunctival macrophages were activated in situ by subconjunctival injection of liposomes containing Con A-activated spleen cell culture supernatants. Control liposomes were loaded with phosphate-buffered saline (PBS). Macrophages exposed to supernatants from Con A-stimulated spleen cells produced 4-fold-higher amounts of NO than unstimulated macrophages. Activation of macrophages via subconjunctival injection of liposomes containing supernatants from Con A-stimulated spleen cell cultures resulted in rapid resolution of the corneal infection. Approximately 80% of animals treated with PBS-containing liposomes demonstrated evidence of corneal disease at day 14 compared to 10% incidence of infection in the Con A-treated group. Moreover, at all time points examined, the clinical appearance of the keratitis in animals treated with liposomes containing Con A supernatant was significantly reduced compared to the group treated with liposomes containing PBS (P < 0.05). Macrophages stimulated with IFN-gamma-containing supernatants killed significant numbers of the trophozoites in vitro (P < 0.05). Killing was inhibited by cytochalasin D, but not by L-N6-1-iminoethyl-L-lysine dihydrochloride (L-NIL), which is a selective inhibitor of inducible NO synthase (INOS).     

Computational consequences of experimentally derived spike-time and weight dependent plasticity rules

We present two weight- and spike-time dependent synaptic plasticity rules consistent with the physiological data of Bi and Poo (J Neurosci 18:10464–10472, 1998). One rule assumes synaptic saturation, while the other is scale free. We extend previous analyses of the asymptotic consequences of weight-dependent STDP to the case of strongly correlated pre- and post-synaptic spiking, more closely resembling associative learning. We further provide a general formula for the contribution of any number of spikes to synaptic drift. Asymptotic weights are shown to principally depend on the correlation and rate of pre- and post-synaptic activity, decreasing with increasing rate under correlated activity, and increasing with rate under uncorrelated activity. Spike train statistics reveal a quantitative effect only in the pre-asymptotic regime, and we provide a new interpretation of the relation between BCM and STDP data.     

Modified AHP-based decision-making model toward accurate selection of eligible maintenance media for production of <Emphasis Type="Italic">taxanes</Emphasis> in <Emphasis Type="Italic">Taxus baccata</Emphasis> callus culture

Attempts were made here to apply a modified analytic hierarchy process (AHP) approach based on refinement assay of dominated alternatives in monitoring the most reliable callus maintenance media (supplemented with l-glutamine and Casamino acid) of Taxus baccata callus cultures in terms of five criteria. Generally, regarding stem-derived calli, 6 out of 18 maintenance media were nominated as non-dominated alternatives, and following AHP ranking test Casamino acid-based media (i.e., A12, A15 and A19) were overall nominated as the premiere. Taking leaf-derived calli into account, only l-glutamine-based media in an ascending order of A8, A4, A6, A5, A9 and A3 were introduced as non-dominated alternatives. Such results connote that l-glutamine-based feeding appears to generate more significant results either for continuous calli growth or taxanes production. In contrast, regarding the second explant, stem, both amino acid supplies had fairly equal worth. Our findings, overall, demonstrate promising applications of the proposed AHP method regarding accurate selection of the best callus maintenance cultures of T. baccata for production of different taxanes including paclitaxel, Baccatin III and 10-deacetylbaccatin III. Similarly, this statistical approach could be also applicable for other crops, for instance, for accurate selection of the best callus cultures/media and consequently production improvement of a given plant secondary metabolite/product.     

Impact of single-site axonal GABAergic synaptic events on cerebellar interneuron activity

Axonal ionotropic receptors are present in a variety of neuronal types, and their function has largely been associated with the modulation of axonal activity and synaptic release. It is usually assumed that activation of axonal GABA_ARs comes from spillover, but in cerebellar molecular layer interneurons (MLIs) the GABA source is different: in these cells, GABA release activates presynaptic GABA_A autoreceptors (autoRs) together with postsynaptic targets, producing an autoR-mediated synaptic event. The frequency of presynaptic, autoR-mediated miniature currents is twice that of their somatodendritic counterparts, suggesting that autoR-mediated responses have an important effect on interneuron activity. Here, we used local Ca²⁺ photolysis in MLI axons of juvenile rats to evoke GABA release from individual varicosities to study the activation of axonal autoRs in single release sites. Our data show that single-site autoR conductances are similar to postsynaptic dendritic conductances. In conditions of high [Cl⁻]_i, autoR-mediated conductances range from 1 to 5 nS; this corresponds to ∼30–150 GABA_A channels per presynaptic varicosity, a value close to the number of channels in postsynaptic densities. Voltage responses produced by the activation of autoRs in single varicosities are amplified by a Na_v-dependent mechanism and propagate along the axon with a length constant of 91 µm. Immunolabeling determination of synapse location shows that on average, one third of the synapses produce autoR-mediated signals that are large enough to reach the axon initial segment. Finally, we show that single-site activation of presynaptic GABA_A autoRs leads to an increase in MLI excitability and thus conveys a strong feedback signal that contributes to spiking activity.     

Adipose tissue is a regulated source of interleukin-10

White adipose tissue (WAT) is the source of pro- and anti-inflammatory cytokines and we have recently shown that this tissue is a major source of the anti-inflammatory interleukin (IL)-1 receptor antagonist (IL-1Ra). We now aimed at identifying additional adipose-derived cytokines, which might serve as regulators of IL-1Ra. We demonstrate here for the first time that the antiinflammatory cytokine IL-10 is secreted by human WAT explants and that it is up-regulated by LPS and TNF-alpha in vitro, as well as in obesity in humans (2- and 6-fold increase in subcutaneous and visceral WAT, respectively) and rodents (4-fold increase).     

The Role of Secretory Immunoglobulin A in the Natural Sensing of Commensal Bacteria by Mouse Peyer's Patch Dendritic Cells

The mammalian gastrointestinal (GI) tract harbors a diverse population of commensal species collectively known as the microbiota, which interact continuously with the host. From very early in life, secretory IgA (SIgA) is found in association with intestinal bacteria. It is considered that this helps to ensure self-limiting growth of the microbiota and hence participates in symbiosis. However, the importance of this association in contributing to the mechanisms ensuring natural host-microorganism communication is in need of further investigation. In the present work, we examined the possible role of SIgA in the transport of commensal bacteria across the GI epithelium. Using an intestinal loop mouse model and fluorescently labeled bacteria, we found that entry of commensal bacteria in Peyer''s patches (PP) via the M cell pathway was mediated by their association with SIgA. Preassociation of bacteria with nonspecific SIgA increased their dynamics of entry and restored the reduced transport observed in germ-free mice known to have a marked reduction in intestinal SIgA production. Selective SIgA-mediated targeting of bacteria is restricted to the tolerogenic CD11c⁺CD11b⁺CD8⁻ dendritic cell subset located in the subepithelial dome region of PPs, confirming that the host is not ignorant of its resident commensals. In conclusion, our work supports the concept that SIgA-mediated monitoring of commensal bacteria targeting dendritic cells in the subepithelial dome region of PPs represents a mechanism whereby the host mucosal immune system controls the continuous dialogue between the host and commensal bacteria.     

Concurrent occurrence of α-amylase inhibitor and stimulator in red kidney bean seed: physiological implications

It is hypothesized that since protein α-amylase inhibitor (α-AI) and stimulator might be present together in red kidney bean (Phaseolus vulgaris L.) seeds, their in vitro interactions might influence their detection and quantification. Assay of α-AI using extracts from the embryonic axes revealed an unexpected finding in that the extracts stimulated rather than inhibited α-amylase activity. The cotyledon extracts exhibited inhibitory or enhancement effect on α-amylase activity depending on whether prior to the α-amylase assay they had been boiled for 10 min or not. Phytohemagglutinin (PHA-L in particular) is implicated in the present study as a stimulator of α-amylase activity co-extracted with α-AI from red kidney bean cotyledons. The importance of these findings is discussed in relation to the possible widespread occurrence of protein α-amylase stimulator in seeds and other plant parts.