The effects of diet mixing on consumer fitness: macroalgae, epiphytes, and animal matter as food for marine amphipods

Herbivores are thought to achieve adequate nutrition by consuming numerous species of plants or by occasionally consuming animal tissue. Although active selection of diverse foods is common in nature, the relationship between diet mixing and consumer fitness is poorly understood, especially in marine environments. We studied the fitness-based consequences of dietary mixing in the sympatric amphipods Ampithoe marcuzzii, A. valida, Cymadusa compta, and Gammarus mucronatus by measuring survivorship, growth, and fecundity of these amphipods when they were offered single species of algae, a single animal food, a mixture of algal species, or a combination of algae and animal matter. For the more sedentary, tube-building amphipods A. marcuzzii, A. valida, and C. compta, fitness on mixed algal diets was matched by fitness on at least one of the monospecific algal diets, suggesting that they could benefit from preferential feeding on those algae in the field. The more mobile amphipod, G. mucronatus, survived and grew similarly on the mixed diets and on the filamentous brown alga Ectocarpus siliculosus. However, its fecundity was significantly higher when feeding on the algal and animal mixture than on Ectocarpus alone. Additionally, for G. mucronatus, fitness on mixed algae, mixed algae plus animal matter, and animal matter alone was equivalent, although female growth (but not gonad production) was slightly lower on animal matter alone than on the mixed algae combined with animal food. Thus the more mobile amphipod, G. mucronatus, was the only species able to perform well on animal food alone. In contrast, A. valida and C. compta experienced large negative effects when limited to consuming animal matter alone. For these two species, combining algae and animal matter did not enhance fitness over combining only algae. Fitness effects of specific algal diets showed some general similarities, but also considerable variance among the amphipods. For example, E. siliculosus was generally better food than other algae for all four amphipods, whereas Sargassum filipendula was generally poor. However, A. marcuzzii did not suffer negative effects of consuming only Sargassum. The red alga Polysiphonia sp. and the green alga Enteromorpha flexuosa decreased fitness in A. marcuzzii, C. compta, and G. mucronatus, but not A. valida, and the negative effects of Polysiphonia were considerably larger for A. marcuzzii than for the other amphipods. Our data show that nutritional requirements, even among related species (e.g., A. marcuzzii and A. valida), can be dramatically different. Diet mixing may benefit more mobile consumers like Gammarus that are better able to search for different foods, and may be less important for more sedentary herbivores like Ampithoe and Cymadusa that consume, and live in close association with, individual host plants. Received: 14 September 1998 / Accepted: 22 October 1999     

Quantification of nuclear DNA and G-C content in marine macroalgae by flow cytometry of isolated nuclei

Summary The amounts of nuclear DNA in ten species of seaweeds belonging to the Rhodophyceae, Phaeophyceae, and Chlorophyceae were determined by flow cytometric analysis of nuclei isolated from protoplasts. Genome size was determined from the fluorescence of the nuclei stained with ethidium bromide. The size of the nuclear genome ranged from 0.13 pg per cell in the 1 C population ofUlva rigida to 3.40 pg per cell in the 2 C population ofSphacelaria sp. GC% analysis was based on staining with either Hoechst 33342 or mithramycin A, two fluorochromes specific for the bases A-T and G-C, respectively. Two models were used for the estimation of the proportion of guanine plus cytosine in the nuclear genome. The first one was based on the linear relationships mithramycin A fluorescence/G-C content and ethidium bromide fluorescence/total DNA content. The second model, based on the curvilinear relationships Hoechst 33342 fluorescence/A-T content and mithramycin A fluorescence/G-C content, resulted in comparatively more homogenous and consistent data and appears more accurate. Comparison with previous reports from other methods for the physical investigation of nuclear genomes shows that flow cytometry of nuclei isolated from protoplasts is an accurate, convenient and robust technique to assay for genome sizes and base pair composition in marine macroalgae.Abbreviations A-T nucleic bases adenine and thymine - CRBC chicken red blood cell - FALS forward-angle light scatter - G-C nucleic bases guanine and cytosine - SEIM sorbitol enzymatic incubation medium - SWIM sea water incubation medium - Tm thermal denaturation temperature of DNA     

Effects of height on the shore and complexity of habitat on abundances of amphipods on rocky shores in New South Wales, Australia

Patterns of abundance of epifaunal crustaceans were compared between two common brown algae in intertidal rock pools on a shore near Sydney, New South Wales, Australia. Amphipods were significantly more abundant on Sargassum globulariaefolium (which was most common low on the shore) than on Hormosira banksii (more common at mid-shore levels). Experiments tested the hypotheses that height on the shore and structural complexity of the algae were the main factors influencing patterns of abundance of amphipods. Reciprocal transplants using defaunated plants and similar experiments using artificial plants allowed simultaneous tests of these hypotheses. The abundance of a common intertidal amphipod, Sunampithoe graxon was significantly smaller on Sargassum transferred from low to mid-tidal level of the shore than on any other treatments, indicating that height on the shore affects numbers of this species. There were no Sunampithoe on Hormosira even when transplanted to low shore areas, indicating that other factors, apart from the height on the shore, are also important for this amphipod. Another common amphipod, Hyale maroubrae was generally found in greatest abundances on Hormosira transplanted from mid- to low shore and on Sargassum low on the shore, again indicating the importance of height on the shore. There was no difference in abundance of amphipods on ‘complex’ and ‘simple’ artificial plants indicating that structural complexity (as defined in this study) did not influence these amphipods.     

Distinct roles for hindbrain and paraxial mesoderm in the induction and patterning of the inner ear revealed by a study of vitamin-A-deficient quail

The hindbrain and cranial paraxial mesoderm have been implicated in the induction and patterning of the inner ear, but the precise role of the two tissues in these processes is still not clear. We have addressed these questions using the vitamin-A-deficient (VAD) quail model, in which VAD embryos lack the posterior half of the hindbrain that normally lies next to the inner ear. Using a battery of molecular markers, we show that the anlagen of the inner ear, the otic placode, is induced in VAD embryos in the absence of the posterior hindbrain. By performing grafting and ablation experiments in chick embryos, we also show that cranial paraxial mesoderm which normally lies beneath the presumptive otic placode is necessary for otic placode induction and that paraxial mesoderm from other locations cannot induce the otic placode. Two members of the fibroblast growth factor family, FGF3 and FGF19, continue to be expressed in this mesodermal population in VAD embryos, and these may be responsible for otic placode induction in the absence of the posterior hindbrain. Although the posterior hindbrain is not required for otic placode induction in VAD embryos, the subsequent patterning of the inner ear is severely disrupted. Several regional markers of the inner ear, such as Pax2, EphA4, SOHo1 and Wnt3a, are incorrectly expressed in VAD otocysts, and the sensory patches and vestibulo-acoustic ganglia are either greatly reduced or absent. Exogenous application of retinoic acid prior to 30 h of development is able rescue the VAD phenotype. By performing such rescue experiments before and after 30 h of development, we show that the inner ear defects of VAD embryos correlate with the absence of the posterior hindbrain. These results show that induction and patterning of the inner ear are governed by separate developmental processes that can be experimentally uncoupled from each other.     

The induction of diapause in <Emphasis Type="Italic">Moina</Emphasis> by species-specific chemical cues

The ability to change the reproduction mode and produce diapausing eggs, which is prevalent in many zooplankton species, significantly impacts on the evolution and ecology of aquatic communities. The production of diapausing eggs is controlled by multiple effects of biotic and abiotic factors, including infochemicals. We have investigated the effects of chemicals exuded by conspecifics and ecologically close competing congers, Moina brachiata and M. macrocopa, which coexist in the same water body, and by larger Cladocera species (Daphnia magna) on the change of reproduction mode, specific growth rate and fecundity of M. brachiata and M. macrocopa females. The production of gametogenetic eggs in both species was detected only in waters from crowded cultures of conspecifics. The water from crowded cultures of conspecifics reduced the specific growth rate of the juvenile females of both species that later switched to gametogenesis. While it either did not affect (in M. macrocopa) or even increase (in M. brachiata) the specific growth rate of the juvenile females that later reproduced by parthenogenesis. Females of M. macrocopa released significantly fewer neonates in the water from crowded cultures of conspecifics than in all other treatments, while the fecundity of M. brachiata females was the same in all treatments. To understand the phenomenon of diapause induction under the effect of chemical cues in zooplankton, a link between laboratory tests and ecological research should be established, and the chemical composition of the signals should be determined.     

Intraspecific variation in a generalist herbivore accounts for differential induction and impact of host plant defences

Plants and herbivores are thought to be engaged in a coevolutionary arms race: rising frequencies of plants with anti-herbivore defences exert pressure on herbivores to resist or circumvent these defences and vice versa. Owing to its frequency-dependent character, the arms race hypothesis predicts that herbivores exhibit genetic variation for traits that determine how they deal with the defences of a given host plant phenotype. Here, we show the existence of distinct variation within a single herbivore species, the spider mite Tetranychus urticae, in traits that lead to resistance or susceptibility to jasmonate (JA)-dependent defences of a host plant but also in traits responsible for induction or repression of JA defences. We characterized three distinct lines of T. urticae that differentially induced JA-related defence genes and metabolites while feeding on tomato plants (Solanum lycopersicum). These lines were also differently affected by induced JA defences. The first line, which induced JA-dependent tomato defences, was susceptible to those defences; the second line also induced JA defences but was resistant to them; and the third, although susceptible to JA defences, repressed induction. We hypothesize that such intraspecific variation is common among herbivores living in environments with a diversity of plants that impose diverse selection pressure.     

Differential induction of genes related to solvent formation during the shift from acidogenesis to solventogenesis in continuous culture of Clostridium acetobutylicum

Abstract The expression of all sequenced acetone and butanol formation genes was followed using mRNA analysis during the shift from acidogenesis to solventogenesis in continuous culture of Clostridium acetobutylicum . Differential induction or derepression of the bdhA, bdhB , and adc genes as well as the sol operon was observed during the pH-induced shift. The order of induction of the three different butanol dehydrogenase genes was found to be bdhA-sol operon- bdhB , offering an explanation for the physiological role of the respective enzymes. Peak mRNA synthesis of an autolysin and a heat shock gene at the onset of solventogenesis was detected in addition to the above-mentioned genes. None of the hitherto sequenced genes of butanologenic enzymes was found to be involved in butanol production during the Methyl viologen-induced shift, indicating the presence of yet unknown genes encoding alcohol and aldehyde dehydrogenases.     

Detection of virulent Shigella and enteroinvasive Escherichia coli by induction of the 43 kDa invasion plasmid antigen, ipaC

The invasion plasmid antigen, ipaC (43 kDa) of Shigella spp. and enteroinvasive Escherichia coli (EIEC) could be induced in vitro by growing them in the presence of Congo red. An enzyme-linked immunosorbent assay (ELISA) using antibodies to the 43 kDa protein of Shigella has been developed for specific detection of virulent Shigella spp and EIEC. The test is independent of initial isolation of individual colonies. As few as 10² CFU/ml of virulent Shigella present in mixed cultures could be detected and concurrently their susceptibility to antibiotics could be analysed after an initial growth of 8–16 h in Congo red-containing medium. The test may prove useful in the diagnosis and treatment of bacillary dysentery caused either by Shigella or EIEC through their rapid identification and proper antimicrobial therapy.     

Feeding preferences and food searching strategies mediated by air- and water-borne cues in the mud whelk Terebralia palustris (Potamididae: Gastropoda)

The gastropod T. palustris is one of the major species responsible for leaf consumption and degration within the Indo-Pacific mangrove forests, and it strongly competes with herbivorous sesarmid crabs in consuming fallen leaves. This snail feeds at high and low tides and it is able to locate food items by means of chemical cues. The aim of this study was to assess the food preferences of T. palustris and to define its feeding strategies at low and high tides, by conducting field trials on water-borne mediated food location at high tide, grazing rate and the chemical attraction exerted by different mangrove leaves. The results showed that T. palustris was able to perceive underwater grazed leaves. In addition, we demonstrated that T. palustris consumes all the mangrove species (preferentially the Rhizophoraceae leaves) but Xilocarpus granatum. Moreover, this snail is differentially attracted to different mangrove species: the major attractive power is wielded by the rhizophoracean species and Pemphis acidula, while X. granatum does not attract this snail at all. The efficacy and adaptive value of a chemically mediated food searching strategies is unquestionable since by using this ability T. palustris can locate and reach the leaves it preferentially consumes. Moreover, T. palustris is the only macrobenthic species of East Africa mangroves able to search, detect and consume mangrove leaves at both high and low tides. Such an expanded feeding window permits T. palustris to occupy temporal niches left empty by the sesarmid crabs.     

Relative impacts of Daphnia grazing and direct stimulation by fish on phytoplankton abundance in mesocosm communities

• 1 Planktivorous fish were hypothesised to influence the abundance of algal biomass in lakes by changing zooplankton grazing, affecting zooplankton nutrient recycling and by direct recycling of nutrients to phytoplankton. The relative roles of direct fish effects vs. zooplankton grazing were tested in mesocosm experiments by adding to natural communities large grazing zooplankton (Daphnia carinata) and small planktivorous fish (mosquitofish or juveniles of Australian golden perch).
• 2 The addition of Daphnia to natural communities reduced the numbers of all phytoplankton less than 30 µm in size, but did not affect total biomass of phytoplankton as large Volvox colonies predominated.
• 3 The addition of Daphnia also reduced the abundance of some small (Moina, Bosmina, Keratella) and large (adult Boeckella) zooplankton, suggesting competitive interactions within zooplankton.
• 4 The addition of mosquitofish to communities containing Daphnia further reduced the abundance of some small zooplankton (Moina, Keratella), but increased the numbers of Daphnia and adult Boeckella. In spite of the likely increase in grazing due to Daphnia, the abundance of total phytoplankton and dominant alga Volvox did not decline in the presence of mosquitofish but was maintained at a significantly higher level than in control.
• 5 The addition of juveniles of golden perch to communities containing Daphnia reduced the abundance of small zooplankton (Moina), increased the abundance of large zooplankton (adult Boeckella) but had no significant effect on Daphnia and total phytoplankton abundance.
• 6 The results of the present study suggest that some planktivorous fish can promote the growth of phytoplankton in a direct way, probably by recycling nutrients, and even in the presence of large grazers. However, the manifestation of the direct effect of fish can vary with fish species.

     

Synergistic induction of hepatocyte growth factor in human skin fibroblasts by the inflammatory cytokines interleukin-1 and interferon-gamma

Hepatocyte growth factor (HGF) is one of the vital factors for wound healing. HGF expression markedly increases in wounded skin and is mainly localized in dermal fibroblasts. HGF expression level in human dermal fibroblasts in vitro, however, is low and thus may be stimulated by some factors in the process of wound healing. Candidates of the factors are inflammatory cytokines released by polymorphonuclear and mononuclear cells infiltrating the wounded area, but HGF production in human dermal fibroblasts is only slightly induced by interleukin (IL)-1, tumor necrosis factor (TNF)-alpha or interferon (IFN)-gamma. We here report that a combination of IL-1beta and IFN-gamma or a combination of TNF-alpha and IFN-gamma very markedly induced HGF production. The synergistic effect of the former was more marked than that of the latter. Synergistic effects of IL-1beta and IFN-gamma were observed at more than 10 pg/ml and 10 IU/ml, respectively, and were detectable as early as 12 h after addition. Neither IFN-alpha nor IFN-beta was able to replace IFN-gamma. HGF mRNA expression was also synergistically upregulated by IL-1beta and IFN-gamma. IL-1beta plus IFN-gamma-induced synergistic production of HGF was potently inhibited by treatment of cells with the extracellular signal-regulated kinase (ERK) kinase inhibitor PD98059 and the p38 inhibitor SB203580 but not by the c-Jun N-terminal kinase (JNK) inhibitor SP600125. Taken together, our results indicate that a combination of IL-1beta and IFN-gamma synergistically induced HGF production in human dermal fibroblasts and suggest that activation of ERK and p38 but not of JNK is involved in the synergistic effect.     

Pathogenic bacteria prime the induction of Toll-like receptor signalling in human colonic cells by the Gal/GalNAc lectin Carbohydrate Recognition Domain of Entamoeba histolytica

In mixed intestinal infections with Entamoeba histolytica trophozoites and enteropathogenic bacteria, which are wide-spread in areas of endemic amoebiasis, interaction between the pathogens could be an important factor in the occurrence of invasive disease. It has been reported that exposure of human colonic cells to enteropathogenic bacteria increased trophozoite adherence to the cells and their subsequent damage. We report here that the Carbohydrate Recognition Domain (CRD) of the amoebic Gal/GalNAc lectin binds to Toll-like receptors TLR-2 and TLR-4 in human colonic cells, activating the “classic” signalling pathway of these receptors. Activation induced expression of TLR-2 and TLR-4 mRNAs and the mRNAs of pro-inflammatory cytokines, as well as an increase in the corresponding proteins. Direct correlation was observed between the increased expression of TLRs and pro-inflammatory cytokines, the enhanced adhesion of trophozoites to the cells and the inflicted cell damage. When cells were exposed to pathogenic bacteria Staphylococcus aureus (Gram+) or Shigella dysenteriae (Gram−), elements of an innate immune response were induced. CRD by itself elicited a similar cell response, while exposure to a commensal Escherichia coli had a null effect. Pre-exposure of the cells to pathogenic bacteria and then to CRD rendered an inflammatory-like microenvironment that after addition of trophozoites facilitated greater cell destruction. Our results suggest that CRD is recognised by human colonic cells as a pathogen-associated-molecular-pattern-like molecule and as such can induce the expression of elements of an innate immune response. In the human host, an exacerbated inflammatory environment, derived from pathogen interplay, may be an important factor for development of invasive disease.     

Smad2/3 activities are required for induction and patterning of the neuroectoderm in zebrafish

Smad2 and Smad3, two essential nuclear effectors of transforming growth factor (Tgf)-β signals, have been found to be implicated in mesoderm and endoderm development in vertebrate embryos. However, their roles in the induction and patterning of the neuroectoderm are not well established. In this study, we show that interference with Smad2/3 activities in zebrafish embryos, by injecting dnsmad3b mRNA encoding a dominant negative Smad3b mutant, inhibits the expression of the early neural markers sox2 and sox3 at the onset of gastrulation and results in reduction of the anterior neuroectodermal marker otx2 as well as the posterior neuroectodermal marker hoxb1b during late gastrulation, suggesting a role of Smad2/3 activities in neural induction. Conversely, excess Smad2/3 activities, caused by injecting smad3b mRNA, lead to an enhancement of sox2 and sox3 expression in the ventral domains but an inhibition of their expression in the dorsalmost region at early stages. Overexpression of smad3b also causes ventral expansion of the otx2 and hoxb1b expression domains accompanied with rostral shift of the hoxb1b domain at late gastrulation stages. Collectively, these data indicate that Smad2/3 activities are required for neural induction and neuroectodermal posteriorization in zebrafish. Knockdown of chordin partially inhibits effect of smad3b overexpression on neural induction, implying that Smad2/3 exert their effect on neural induction in part by regulating the expression of Bmp antagonists. Furthermore, down-regulation or up-regulation of Smad2/3 activities in MZoep mutant embryos, which lack the organizer and mesendodermal tissues due to deficiency of Nodal signaling, still affects induction and patterning of the neuroectoderm, suggesting that Smad2/3 activities are implicated in neural development in the absence of the organizer and mesendodermal tissues. We additionally demonstrate that Smad2/3 activities cooperate with Wnt and Fgf signals in neural development. Thus, Smad2/3 activities play important roles not only in mesendodermal development but also in neural development during early vertebrate embryogenesis.     

Inhibition of hepatocyte growth factor induction in human dermal fibroblasts by interleukin-1 and its prevention by interferon-gamma

Hepatocyte growth factor (HGF) is one of the vital factors for liver regeneration. HGF production is induced by the activation of protein kinase A and protein kinase C-mediated pathways, interleukin (IL)-1, tumor necrosis factor (TNF)-alpha, and epidermal growth factor (EGF) in mesenchymal cells. We here report that IL-1 and TNF-alpha, hitherto regarded as HGF inducers, potently inhibited HGF production stimulated by other HGF inducers. IL-1alpha, IL-1beta, and TNF-alpha alone had minimal stimulating effects on HGF production in human dermal fibroblasts, but they strongly inhibited production of HGF induced by cholera toxin, 8-bromo-cAMP, EGF, and phorbol 12-myristate 13-acetate (PMA). Moreover, although the high level of HGF production in MRC-5 cells was enhanced by PMA and less markedly by IL-1beta, HGF production in MRC-5 cells treated with PMA plus IL-1beta was less than that in the cells treated with PMA alone. In the presence of interferon (IFN)-gamma, however, cholera toxin- and 8-bromo-cAMP-induced HGF production was not inhibited by IL-1beta. Pretreatment of cells with IL-1beta suppressed the phosphorylation of cAMP-responsive element-binding protein induced by cholera toxin but not that induced by 8-bromo-cAMP. Taken together, our results indicate that IL-1 inhibited HGF production stimulated by various inducers, including protein kinase A-activating agents, and that IFN-gamma overcame this inhibition of induction of HGF production.     

Patterning of an ascidian embryo along the anterior-posterior axis through spatial regulation of competence and induction ability by maternally localized PEM

In ascidian embryos, anterior-posterior (A-P) patterning of the vegetal cells is regulated by posteriorizing activities of a localized egg region known as posterior-vegetal cortex/cytoplasm (PVC). PEM is an essential component of the PVC and is involved in the posterior-specific cell cleavage pattern. Here we report a novel function of PEM independently of its function in cleavage regulation; it controls cell fate by excluding competence to respond to the FGF signal for notochord induction from posterior-vegetal cells. PEM was found to regulate the nuclear accumulation of β-catenin, an upstream activator of the competence factor. PEM also influences A-P patterning in the animal hemisphere. It was found to regulate FGF signal expression and restrict the occurrence of brain induction only in the anterior region. Our results suggest a model in which PEM patterns the embryo along the A-P axis through regulation of the spatial distribution of competence and induction ability.     

The role of major histocompatibility complex expression on head and neck cancer cells in the induction of autologous cytotoxic T lymphocytes

Using head and neck tumors, we studied the role of HLA class I and DR antigens on tumor cells in cytotoxic T lymphocyte (CTL) induction. Expression of major histocompatibility complex (MHC) antigens was investigated by two-color flow cytometry analysis and for this study we used the tumor cells, over 50% of which expressed both HLA class I and DR antigens on their surface. In seven cases, tumor cells were divided into three groups according to the specificity of monoclonal antibodies (mAb) to MHC to study the role of MHC antigens on tumor cells in CTL induction: one was not blocked (MHC double-positive tumor), a second was blocked by anti-class I mAb (class-Ingative DR-positive tumor) and third was blocked by anti-DR mAb (class-I-positive DR-negative tumor). Subsequently, these tumors were used to stimulate an autologous mixed lymphocyte/tumor cell culture for 5 days (MLTC) followed by further cultivation with interleukin-2 for 12 days. The induced autologous tumor killer cells were most cytotoxic when non-treated tumors, which consist mainly of cells that are both HLA-class I and DR-positive, were used as stimulator cells. When the tumor cells blocked by anti-DR mAb were used as stimulators, autologous tumor killer activity was lower than that induced by tumor cells blocked by anti-class-I mAb. Moreover, cytolysis by autologous tumor killer cells induced by stimulation of non-treated tumor cells was blocked during the effector phase, 26.6%–42.3% and 32.7%–53.8% by anti-class-I and anti-DR mAb respectively, suggesting that majority of the autologous tumor killer cells are MHC-restricted CD8⁺ or CD4⁺ CTL. These results suggest that both MHC class I and class II antigens on head and neck tumor cells play a critical role in inducing CTL.     

Mechanisms of rapid induction of interleukin-22 in activated T cells and its modulation by cyclosporin a

IL-22 is an immunoregulatory cytokine displaying pathological functions in models of autoimmunity like experimental psoriasis. Understanding molecular mechanisms driving IL-22, together with knowledge on the capacity of current immunosuppressive drugs to target this process, may open an avenue to novel therapeutic options. Here, we sought to characterize regulation of human IL22 gene expression with focus on the established model of Jurkat T cells. Moreover, effects of the prototypic immunosuppressant cyclosporin A (CsA) were investigated. We report that IL-22 induction by TPA/A23187 (T/A) or αCD3 is inhibited by CsA or related FK506. Similar data were obtained with peripheral blood mononuclear cells or purified CD3(+) T cells. IL22 promoter analysis (-1074 to +156 bp) revealed a role of an NF-AT (-95/-91 nt) and a CREB (-194/-190 nt) binding site for gene induction. Indeed, binding of CREB and NF-ATc2, but not c-Rel, under the influence of T/A to those elements could be proven by ChIP. Because CsA has the capability to impair IκB kinase (IKK) complex activation, the IKKα/β inhibitor IKKVII was evaluated. IKKVII likewise reduced IL-22 induction in Jurkat cells and peripheral blood mononuclear cells. Interestingly, transfection of Jurkat cells with siRNA directed against IKKα impaired IL22 gene expression. Data presented suggest that NF-AT, CREB, and IKKα contribute to rapid IL22 gene induction. In particular the crucial role of NF-AT detected herein may form the basis of direct action of CsA on IL-22 expression by T cells, which may contribute to therapeutic efficacy of the drug in autoimmunity.     

The significance of the kinetic analysis of fluorescence induction in DCMU-inhibited chloroplasts in terms of photosystem 2 connectivity and heterogeneity

A study has been made on the slow component (β_max) of chlorophyll fluorescence induction curves exhibited by DCMU-inhibited pea thylakoids. In the absence of high levels of screening cations, β_max is high and the fluorescence yield low, while the addition of K⁺, Mg²⁺ or Tris(ethylenediamine) cobaltic(III) cation (TEC³⁺) decreases β_max and increases fluorescence yield. These changes are inhibited when the thylakoids are fixed with glutaraldehyde. By comparing cation- and light-harvesting chlorophyll a/b-protein (LHCP) phosphorylation-induced changes it can be seen that β_max correlates with changes in the overall kinetics of photosystem (PS) 2 photoreduction, as indicated from the normalised area over the induction curve (Anorm). When the plastoquinone (PQ) pool is chemically reduced by dithionite, prior to the initiation of the curve, both F_o and F_m are increased and the slow component removed. These observations are used to question the concept of two structurally distinct PS2 centres [Arch. Biochem. Biophys. (1978) 190, 523–530] and to discuss the use of β_max in monitoring PS2 organisation.