Ecophysiological traits explain species dominance patterns in macroalgal blooms

Macroalgal blooms are a growing environmental problem in eutrophic coastal ecosystems world wide. These blooms are dominated typically by only one out of several co-occurring opportunistic species, which are all favored by increased nutrient loads. We asked whether pronounced dominance of filamentous Pilayella littoralis Kjellm. (Phaeophyceae) over foliose Enteromorpha intestinalis L. (Chlorophyceae) in the Baltic Sea can be explained by interspecific physiological differences. In laboratory experiments, we analyzed uptake kinetics of nitrate, ammonium, and phosphate and the time dependency of uptake rates for both species. We further examined growth rates and nutrient assimilation in relation to single and combined enrichment with nitrate and phosphate, and three different nitrogen sources. Overall, we did not detect distinct differences in uptake, growth, and assimilation rates between P. littoralis and E. intestinalis. Minor differences and the related advantages for single species are discussed. Highest maximal uptake rates were found for ammonium, followed by nitrate and phosphate. Strong time dependency of uptake occurred, with the highest rates during the first 15 to 30 min. Nitrate enrichment had far more of an effect on growth than phosphate. Enrichment with urea, ammonium, and nitrate significantly increased growth rates without interspecific differences. A larger surface area to volume (SA/V) ratio in Pilayella compared with Enteromorpha did not translate into greater physiological capacity. We conclude that species dominance patterns in macroalgal blooms are not always a direct result of different ecophysiological traits among species. Ecological traits such as susceptibility to herbivory are important factors in determining species distribution in the field.     

Propagule banks, herbivory and nutrient supply control population development and dominance patterns in macroalgal blooms

Destructive macroalgal mass blooms threaten estuarine and coastal ecosystems worldwide. We asked which factors regulate macroalgal bloom intensity, distribution and species composition. In field experiments in the Baltic Sea, we analyzed the relative effects of nutrients, herbivores and algal propagule banks on population development and dominance patterns in two co-occurring bloom-forming macroalgae, Enteromorpha intestinalis and Pilayella littoralis . Both species were highly affected by the combined effects of a propagule bank, herbivory and nutrients. The magnitude of effects varied with season. The propagule bank was an important overwintering mechanism for both algae, and allowed for recruitment two months earlier than recruitment via freshly dispersed propagules. This provided a seasonal escape from intense herbivory and nutrient limitation later in the year. Favored by massive recruitment from the propagule bank, Enteromorpha was the superior space occupier in early spring, thereby reducing recruitment of Pilayella . Elimination of the propagule bank and recruitment via freshly dispersed propagules favored Pilayella . Strong and selective herbivory on Enteromorpha supported Pilayella in the presence, but not in the absence of the propagule bank. Nutrient enrichment in summer counteracted herbivore pressure on Enteromorpha , thereby negatively affecting Pilayella . Herbivore and nutrient effects were more pronounced for early life stages than adult algae. These results show that recruitment processes and forces affecting early life stages at the beginning of the vegetation period determine development and dominance patterns of macroalgal blooms. Herbivores naturally suppress blooms but increasing nutrient enrichment can override this important control mechanism. The propagule bank plays a previously unrecognized role for population and community dynamics.     

Control of macroalgal blooms at early developmental stages: Pilayella littoralis versus Enteromorpha spp.

Although blooms of opportunistic fast-growing macroalgae now occur frequently in coastal ecosystems affected by eutrophication, their initiation and control is little understood. Most previous studies have focused on the ecophysiology of adult algae only. We show that spores and/or germlings may represent critical stages in the life cycles and mass-developments of co-occurring opportunistic macroalgae in the Baltic (Pilayella littoralis and Enteromorpha spp.). We investigated the overwintering of spores, timing of germination, subsequent growth, and grazing on spores and germlings, in order to explain the initiation of mass blooms and species dominance patterns. In the field, Enteromorpha spp. showed 10- to 50-fold higher abundances of overwintering microscopic forms (up to 330 individuals cm⁻²) than P. littoralis. Moreover, we found continuous production of spores (up to 1.2 million settling spores m⁻² h⁻¹) from April to October in Enteromorpha spp., while there was evidence of only a short reproductive period in Pilayella. However, in spring, germlings and adults of P. littoralis appeared earlier in the field and reached a 10-fold higher biomass than Enteromorpha spp. In factorial laboratory experiments including temperature and light, there were clear differences in timing of germination. P. littoralis germinated at 5°C whereas Enteromorpha spp. required temperatures of 10–15°C for germination. In contrast, we detected only minor differences in growth response among adults of P. littoralis and Enteromorpha spp. Germination, not growth of adults, appeared to be the ecophysiological bottleneck for initiating mass spring development. Following the spring Pilayella bloom, Enteromorpha germlings occurred massively in the field (April–September), but rarely developed into adults. In laboratory feeding experiments we tested whether crustacean mesograzers common in summer may control development of Enteromorpha germlings. Both germination of settled spores and growth of germlings were reduced by 93–99% in the presence of grazers (Idotea chelipes and Gammarus locusta). Thus in addition to ecophysiological constraints, grazers, if present, may play a decisive role in the early life stages of macroalgal mass developments. These results mirror patterns of overwintering of seeds, germination control, seed and seedling predation in terrestrial plant communities. Received: 7 March 1998 / Accepted: 18 November 1998     

Expression of androgen-binding protein (ABP) in human cardiac myocytes.

Cardiomyocytes are known to be androgen targets. Changing systemic steroid levels are thought to be linked to various cardiac ailments, including dilated cardiomyopathy (DCM). The mode of action of gonadal steroid hormones on the human heart is unknown to date. In the present study, we used high-resolution immunocytochemistry on semithin sections (1 microm thick), IN SITU hybridization, and mass spectrometry to investigate the expression of androgen-binding protein (ABP) in human myocardial biopsies taken from male patients with DCM. We observed distinct cytoplasmic ABP immunoreactivity in a fraction of the myocytes. IN SITU hybridization with synthetic oligonucleotide probes revealed specific hybridization signals in these cells. A portion of the ABP-positive cells contained immunostaining for androgen receptor. With SELDI TOF mass spectrometry of affinity purified tissue extracts of human myocardium, we confirmed the presence of a 50 kDa protein similar to ABP. Our observations provide evidence of an intrinsic expression of ABP in human heart. ABP may be secreted from myocytes in a paracrine manner perhaps to influence the bioavailabity of gonadal steroids in myocardium.     

Administration of recombinant IL-4 to humans regulates gene expression, phenotype, and function in circulating monocytes.

As a multifunctional cytokine that can augment certain T cell responses, IL-4 is being evaluated currently as a possible therapeutic agent in the treatment of cancer patients. In this report, PBMC were isolated from such patients before and after IL-4 therapy and were analyzed for phenotypic and functional alterations. Differential blood counts showed that the relative percentages of lymphocytes and, to a lesser degree, monocytes decreased after treatment with IL-4. However, when phenotypically analyzed by FACS, monocyte numbers generally increased, whereas there was a decrease in lymphocyte numbers. Monocytes taken from patients before therapy and cultured with and without LPS exhibited normal patterns of monokine-specific (IL-1 beta and TNF-alpha) mRNA expression, as well as secretion of peptide. The addition of rIL-4 to these cultures, however, resulted in the down-regulation of both gene expression and peptide release. Although monocytes taken from post-therapy patients also displayed normal patterns of monokine gene expression and cytokine release after stimulation with LPS, they were no longer inhibited by the addition of exogenous rIL-4 in vitro. These data suggest that monocytes become refractory to the inhibitory effects of IL-4 after in vivo exposure to this cytokine. However, when these monocytes were examined for expression of CD14, CD32, and CD64, exogenous IL-4 was observed to decrease markedly the appearance of these markers, regardless of whether the cells were obtained before or after therapy. Furthermore, monocytes from post-therapy patients exhibited reduced production of PGE2 and superoxide anion, compared with cells obtained before therapy. This effect persisted in culture independent of the further addition of exogenous IL-4. These data suggest a dichotomy between the IL-4-dependent mechanisms that regulate monokine production and those that regulate certain other monocyte functions. The exact mechanisms that govern these two pathways are not known. These results have important clinical implications for the use of IL-4 as an immunotherapeutic agent, as well as providing insight into the physiologic role of IL-4.     

Host immune response in renal cell cancer: Interleukin-4 (IL-4) and IL-10 mRNA are frequently detected in freshly collected tumor-infiltrating lymphocytes

Human renal cell cancer (RCC) is clearly responsive to immunotherapy. Clinical responses may be mediated by non-specific (e. g. natural killer, NK, cells) or specific MHC-class-I-restricted tumor-specific CD8⁺ T lymphocytes. Typically RCC progresses, however, despite significant infiltration of various lymphoid cells. We examined freshly isolated RCC tumor-infiltrating lymphocytes (TIL) derived from seven RCC patients for cytokine expression by the polymerase chain reaction (PCR). Established RCC tumor cell lines derived from these RCC patients were negative for interleukin-2 (IL-2), IL-4, IL-10, and interferon and found to be positive for tumor necrosis factor (TNF), IL-6, IL-1, granulocyte/macrophage-colony-stimulating factor (GM-CSF), and transforming growth factor 1 (TGF1) message as detected by PCR. An identical pattern of cytokine mRNA expression was identified in other long-term RCC lines and in normal human kidney cells upon culture, but not in two Wilms tumor cell lines tested. Short-term-, and long-term-established RCC lines, but not Wilms tumor lines, secreted substantial levels of GM-CSF, TNF, IL-1, and IL-6 as detected by enzyme-linked immunosorbent assay. Both RCC lines and Wilms tumor lines secreted TGF1. In comparison, normal kidney cells secreted IL-6 and GM-CSF, but not IL-1, or TFG1 under identical in vitro cell culture conditions. We applied PCR-based methods to characterize the cytokine mRNA expression pattern in immune cells infiltrating into renal cell cancer without the need for expansion of such effector cells in vitro. Examining freshly collected RCC TIL by PCR from patients with primary cell cell cancer, we could demonstrate that such cells, but not lympho-mononuclear cells harvested from normal human kidney tissue, typically exhibit IL-4 and IL-10 mRNA expression.     

Interleukin-3 induces proliferation but not lymphokine activated killer activity from human and murine mononuclear cells

Recombinant IL-3 (rIL-3) is a potent colony stimulating factor capable of stimulating early hematopoietic pluripotential progenitor cells and of supporting the differentiation of multiple cells. IL-3 has also been shown to have effects on mature, differentiated circulating cells including eosinophils and T cells. We evaluated the role of exogenous rIL-3 in the generation of cells with LAK activity from murine splenocytes and human bone marrow, spleen, unseparated PBMC and purified null cell preparations. rIL-3 was unable to generate lytic activity from any of these populations by itself and appeared to decrease LAK activity in bone marrow cultures containing high dose IL-2, (bone marrow derived cells (n = 3) with LAK activity for fresh tumor, mean lytic units(LU) 94.6 +/- 63.5 vs 32.8 +/- 44.8 for IL-2 and IL-2 plus IL-3 cultures, respectively p2 less than 0.05). Unlike previous reports testing murine cells, IL-3 priming and subsequent culture in IL-2 of human unseparated bone marrow cells or human or murine splenocytes, failed to generate long-term cultures with lytic activity. IL-3 did, however, induce a dose dependent stimulation of bone marrow and null cell preparations (mean null cell stimulation (3H Thymidine incorporation) with IL-3, 436 +/- 168 cpm vs 9802 +/- 9799 cpm, for 0 vs 10(3) units of IL-3, respectively n = 4, p2 less than 0.05). Furthermore, in bone marrow, unseparated PBMC and null cell cultures, the addition of rIL-3 generated characteristic large blastic appearing cells with prominent basophilic granules.(ABSTRACT TRUNCATED AT 250 WORDS)     

IL-6/IFN-beta-2 as a circulating hormone. Induction by cytokine administration in humans

IL-6/IFN-beta 2 is a family of phosphoglycoproteins ranging in size from 19 to 30 kDa which elicits a broad range of physiologic and immune responses. Several cytokines, including TNF, have been shown to stimulate IL-6 production in cell culture. In this report, we describe the rapid induction of circulating biologically active IL-6 by the systemic administration of rTNF to patients with cancer. Low levels of IL-6 activity could be detected in the sera of patients as early as 5 min after rTNF infusion. IL-6 levels peaked approximately 2 to 3 h after rTNF bolus administration and were undetectable in most cases within 8 h. IL-6 was detected in two separate bioassays--the hybridoma B9 proliferation and the hepatocyte-stimulating factor assay. Maximum detectable levels of IL-6 ranged from 160 to 310 hybridoma growth factor units and 11-82 ng/ml in the hepatocyte-stimulating factor assay. IL-6 induction decreased after serial, daily doses of rTNF. Serial serum samples of patients receiving IL-2 or IFN-alpha were also assayed for IL-6 production. IL-2-treated but not IFN-alpha-treated patients generated low levels of IL-6 (range less than 20 to 95 hybridoma growth factor units/ml). Interestingly, in patients treated with IL-2, serum levels of TNF were detectable and peak TNF activity preceded measurable IL-6 levels. Serum levels of acute phase plasma proteins and of corticosteroid rose in response to rTNF administration. C-reactive protein increased (2.5 to 4.0-fold) within 8 h of rTNF administration and cortisol levels rose (10- to 20-fold) within 4 h after rTNF injection. We conclude that rTNF administration in man leads to the induction of circulating IL-6 which, due to its broad range of activities, may be an important physiologic signal regulating the immune response.     

Structural Degradation in Mediterranean Sea Food Webs: Testing Ecological Hypotheses Using Stochastic and Mass-Balance Modelling

Human-mediated disturbances such as fishing, habitat modification, and pollution have resulted in significant shifts in species composition and abundance in marine ecosystems which translate into degradation of food-web structure. Here, we used a comparative ecological modelling approach and data from two food webs (North-Central Adriatic and South Catalan Sea) and two time periods (mid-late 1970s and 1990s) in the Mediterranean Sea to evaluate how changes in species composition and biomass have affected food-web properties and the extent of ecosystem degradation. We assembled species lists and ecological information for both regions and time periods into stochastic structural and mass-balance food-web models, and compared the outcomes of 22 food-web properties. Our results show strong similarities in structural food-web properties between the North-Central Adriatic and South Catalan Seas indicating similar ecosystem structure and levels of ecological degradation between regions and time periods. In contrast, a comparison with other published marine food webs (Caribbean, Benguela, and US continental shelf) suggested that Mediterranean webs are in an advanced state of ecological degradation. This was reflected by lower trophic height, linkage density, connectance, omnivory, species involved in looping, trophic chain length and fraction of biomass at higher trophic levels, as well as higher generality and fraction of biomass at lower trophic levels. An analysis of robustness to simulated species extinction revealed lower robustness to species removals in Mediterranean webs and corroborated their advanced state of degradation. Importantly, the two modelling approaches used delivered comparable results suggesting that they both capture fundamental information about how food webs are structured. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Imputation of missing genotypes: an empirical evaluation of IMPUTE

Background

Polymorphism in genes of regulating enzymes, transporters and receptors of the neurotransmitters of the central nervous system have been associated with altered behaviour, and single nucleotide polymorphisms (SNPs) represent the most frequent type of genetic variation. The serotonin and dopamine signalling systems have a central influence on different behavioural phenotypes, both of invertebrates and vertebrates, and this study was undertaken in order to explore genetic variation that may be associated with variation in behaviour.

Results

Single nucleotide polymorphisms in canine genes related to behaviour were identified by individually sequencing eight dogs (Canis familiaris) of different breeds. Eighteen genes from the dopamine and the serotonin systems were screened, revealing 34 SNPs distributed in 14 of the 18 selected genes. A total of 24,895 bp coding sequence was sequenced yielding an average frequency of one SNP per 732 bp (1/732). A total of 11 non-synonymous SNPs (nsSNPs), which may be involved in alteration of protein function, were detected. Of these 11 nsSNPs, six resulted in a substitution of amino acid residue with concomitant change in structural parameters.

Conclusion

We have identified a number of coding SNPs in behaviour-related genes, several of which change the amino acids of the proteins. Some of the canine SNPs exist in codons that are evolutionary conserved between five compared species, and predictions indicate that they may have a functional effect on the protein. The reported coding SNP frequency of the studied genes falls within the range of SNP frequencies reported earlier in the dog and other mammalian species. Novel SNPs are presented and the results show a significant genetic variation in expressed sequences in this group of genes. The results can contribute to an improved understanding of the genetics of behaviour.