Differential investment and costs during avian incubation determined by individual quality: an experimental study of the common eider (Somateria mollissima)

Individuals of different quality may have different investment strategies, shaping responses to experimental manipulations, thereby rendering the detection of such patterns difficult. However, previous clutch-size manipulation studies have infrequently incorporated individual differences in quality. To examine costs of incubation and reproductive investment in relation to changes in clutch size, we enlarged and reduced natural clutch sizes of four and five eggs by one egg early in the incubation period in female common eiders (Somateria mollissima), a sea duck with an anorectic incubation period. Females that had produced four eggs (lower quality) responded to clutch reductions by deserting the nest more frequently but did not increase incubation effort in response to clutch enlargement, at the cost of reduced hatch success of eggs. Among birds with an original clutch size of five (higher quality), reducing and enlarging clutch size reduced and increased relative body mass loss respectively without affecting hatch success. In common eiders many females abandon their own ducklings to the care of other females. Enlarging five-egg clutches led to increased brood care rate despite the higher effort spent incubating these clutches, indicating that the higher fitness value of a large brood is increasing adult brood investment. This study shows that the ability to respond to clutch-size manipulations depends on original clutch size, reflecting differences in female quality. Females of low quality were reluctant to increase investment at the cost of lower hatch success, whereas females of higher quality apparently have a larger capacity both to increase incubation effort and brood care investment.     

Mercury Decreases Culturability of <Emphasis Type="Italic">Pseudomonas frederiksbergensis</Emphasis> JAJ 28 in Soil Microcosms

Mercury is a biologically potent heavy metal, which has been found to change the diversity of culturable bacteria. Therefore, we investigated whether Hg kills bacteria in soil or reduces culturability. Soil microcosms were inoculated with Pseudomonas frederiksbergensis JAJ 28 and were sampled regularly during 28 days. The total number of acridine orange-stained cells was relatively constant, and Hg reduced the number on only one sampling day. However, the fraction of culturable cells on 1/10 tryptic soy agar was lowered on days 6, 13, and 21. The number of microcolony forming units, which represents viable cells, was also affected by Hg, but this effect was delayed compared with the effects on CFUs. The amount of headspace CO₂ per cell was overall increased by Hg, another indication of the toxic effects of Hg on the bacterial cells. Our results thus emphasize the need to take culturability into account when studying the effects of heavy metals on bacterial diversity. RID= ID= <E5>Correspondence to:</E5> K. Johnsen: <E5>email:</E5> kjo&commat;vetinst.dk Received: 9 August 2002 / Accepted: 2 October 2002     

The RNA-binding protein SUP-12 controls muscle-specific splicing of the ADF/cofilin pre-mRNA in C. elegans

Tissue-specific alternative pre-mRNA splicing is essential for increasing diversity of functionally different gene products. In Caenorhabditis elegans, UNC-60A and UNC-60B, nonmuscle and muscle isoforms of actin depolymerizing factor (ADF)/cofilin, are expressed by alternative splicing of unc-60 and regulate distinct actin-dependent developmental processes. We report that SUP-12, a member of a new family of RNA recognition motif (RRM) proteins, including SEB-4, regulates muscle-specific splicing of unc-60. In sup-12 mutants, expression of UNC-60B is decreased, whereas UNC-60A is up-regulated in muscle. sup-12 mutations strongly suppress muscle defects in unc-60B mutants by allowing expression of UNC-60A in muscle that can substitute for UNC-60B, thus unmasking their functional redundancy. SUP-12 is expressed in muscle and localized to the nuclei in a speckled pattern. The RRM domain of SUP-12 binds to several sites of the unc-60 pre-mRNA including the UG repeats near the 3'-splice site in the first intron. Our results suggest that SUP-12 is a novel tissue-specific splicing factor and regulates functional redundancy among ADF/cofilin isoforms.     

Changes in pigmentation, bio-optical characteristics and photophysiology, during phytoflagellate succession in mesocosms

Pigmentation, bio-optical characteristics and photophysiology,were studied in mesocosms with different N:P ratios. No significantdifference in biomass or species composition was seen underdifferent nitrogen to phosphorus ratios (N:P), but a temporalsuccession of different flagellate groups was observed in allmesocosms. An initial bloom of prymnesiophytes containing chlorophyll(Chl) c and 19' hexanoyloxyfucoxanthin (19' HOF) was followedby prasinophytes containing Chl b. Electron microscope analysisconfirmed the presence of genera such as Chrysochromulina (Prymnesiophyceae),Tetraselmis and Pyramimonas (Prasinophyceae). Traces of prasinoxanthinin the pigment samples showed that smaller prasinophytes werealso present. Chl b influenced the photophysiology of the prasinophytesresulting in higher Chl a-specific absorption, but a greaterdifference between absorption and scaled fluorescence excitationspectra indicated that light absorbed by Chl b is associatedwith photosystem I (PSI). Since a larger fraction of the lightwas absorbed by chlorophyll in PSI and/or photoprotective carotenoids,the light-saturated Chl a-specific rate of photosynthesis (P^B_m)and maximum light utilization coefficient (^B) decreased when[Chl b] increased. The highest P^B_m values were seen when theratios of fucoxanthins to Chl a were high, indicating that prymnesiophytesmight be more efficient in light harvesting and electron transportthrough photosystem II (PSII) by fucoxanthins and Chl c. Ourresults therefore indicate different light acclimation strategiesin prasinophytes versus prymnesiophytes, which may be reflectedin the successional appearance of these communities in the naturalenvironment. We also suggest that grazing by ciliates and rotiferscaused periodic decreases in phytoplankton biomass, which inturn gave rise to the phytoflagellate succession observed inthe mesocosms.     

Effects of Rhizoctonia infection and drought on peroxidase and chitinase activity in Norway spruce (Picea abies)

Seedlings of Norway spruce were exposed to fungal infection and drought in order to investigate differences in their stress responses on the enzymatic level. Six-week-old seedlings were infected with the root rot fungus Rhizoctonia , or subjected to drought, respectively. Changes at the enzymatic level were more rapid and significantly higher in infected plants in comparison with drought-stressed spruce plants. Rhizoctonia infection resulted in early local and systemic increase in peroxidase and chitinase activity. The most prominent isoforms responding were highly basic peroxidases and chitinases (pI 9–9.5) and several acidic chitinases (pI3–4). An increased intensity of similar peroxidase isoforms was found in drought-affected plants. Two peroxidase isoforms (with pI < 9) accumulated exclusively in response to drought. These results suggest that at an early stage of infection and drought stress, the two stresses can be distinguished by the temporal appearance and isoform profile of peroxidases and chitinases. Changes in enzyme activity appeared before changes in physiological parameters, thus these isoform profiles could be used as early markers of stress conditions in spruce.     

Impaired circulating myeloid DCs from myeloma patients

BACKGROUND: In clinical trials, cancer patients have received immunotherapy based on DCs generated from leukapheresed blood. It would therefore be an advantage to be able to measure blood levels and estimate the phenotype of DC before leukapheresis, to estimate the yield required for preparation of vaccines, or ex vivo stimulation of T cells for adoptive immunotherapy. METHODS: Recently, circulating lineage negative (Lin-) myeloid DC cells and their precursors have been identified by flow cytometry. We apply this strategy to the screening of blood samples from patients with multiple myeloma, in an attempt to characterize and quantitate the subset. By a direct flow cytometry approach, the blood levels of circulating lineage (CD3, CD19, CD14) negative, CD33++, HLA-DR+ cells were estimated before and following ex vivo cell differentiation, and phenotyped by MAbs with specificity against HLA-DR, HLA-ABC, CD1a, CD11c, CD33, CD40, CD49d, CD49e, CD54, CD80, CD83, and CD86. RESULTS: This study demonstrated that multiple myeloma patients have a 50% reduced blood level of Lin-, CD33++, HLA-DR+ myeloid DC, but a DC-precursor level within normal range. Furthermore, GM-CSF and IL-4 ex vivo stimulated DCs demonstrated an impaired up-regulation of the co-stimulatory molecule CD80 and the adhesion molecule CD54. DISCUSSION: These results may have clinical implications as a predictor for yield and functionality of the harvested DCs to be used in vaccination of myeloma patients.     

CO<Subscript>2</Subscript> enrichment,nitrogen fertilization and development of freezing tolerance in Norway spruce

Plant growth and adaptation to cold and freezing temperatures in a CO₂-enriched atmosphere have received little attention despite the predicted effects of elevated CO₂ on plant distribution and productivity. Norway spruce [Picea abies (L.) Karst.] seedlings from latitudinally distinct seed sources (66°N and 60°N) were grown for one simulated growth season under controlled conditions in an atmosphere enriched in CO₂ (70 Pa) and at ambient CO₂ (40 Pa), combined factorially with low (3.6 mM) or high (15.7 mM) concentrations of nitrogen fertilization. There was a clear difference between the two provenances in height growth, in the timing of bud set, and in freezing tolerance. Nitrogen fertilization increased height growth in both provenances, while CO₂ enrichment stimulated height growth only in the southern provenance. We found no significant effects of elevated CO₂ or nitrogen fertilization on the timing of bud set. During cold acclimation, freezing tolerance increased from –10°C to –35°C, and there was a marked increase in all soluble sugars except inositol. Elevated CO₂ in combination with high nitrogen led to a slight increased freezing tolerance in both provenances during the early stages of cold acclimation. However, towards the end of cold acclimation, elevated CO₂ and high nitrogen led to reduced freezing tolerance in the southern provenance, while elevated CO₂ and low nitrogen reduced freezing tolerance in the northern provenance. These results suggest that CO₂ enrichment influences the development of freezing tolerance, and that these responses differ with available nitrogen and between provenances.     

Kinetic and structural characterization of a two-domain streptokinase: dissection of domain functionality

The mammalian protease plasminogen can be activated by bacterial activators, the three-domain (alpha, beta, gamma) streptokinases and the one-domain (alpha) staphylokinases. These activators act as plasmin(ogen) cofactors, and the resulting complexes initiate proteolytic activity of host plasminogen which facilitates bacterial colonization of the host organism. We have investigated the kinetic mechanism of the plasminogen activation mediated by a novel two-domain (alpha, beta) streptokinase isolated from Streptococcus uberis (Sk(U)) with specificity toward bovine plasminogen. The interaction between Sk(U) and plasminogen occurred in two steps: (1) rapid association of the proteins and (2) slow transition to the active complex Sk(U)-PgA. The complex Sk(U)-PgA converted plasminogen to plasmin with the following parameters: K(m) < or = 1.5 microM and k(cat) = 0.55 s(-)(1). The ability of proteolytic fragments of Sk(U) to activate plasminogen was investigated. Only two C-terminal segments (97-261 and 123-261), which both contain the beta-domain (126-261), were shown to be active. They initiated plasminogen activation in complex with plasmin, but not with plasminogen, and thereby exhibited functional similarity to the staphylokinase. The fusion protein His(6)-Sk(U) (i.e., Sk(U) with a small N-terminal tag) acted exclusively in complex with plasmin as well. These observations demonstrate that (1) the N-terminal alpha-domain, including a native N-terminus, was necessary for "virgin" activation of the associated plasminogen in the Sk(U)-PgA complex and (2) the C-terminal beta-domain of Sk(U) is important for recognition of the substrate in the Sk(U)-PgA complex.     

Phylogenetic relationships within the tropical soft coral genera Sarcophyton and Lobophytum (Anthozoa, Octocorallia)

Abstract. The alcyonacean soft coral genera Sarcophyton and Lobophytum are conspicuous, ecologically important members of shallow reef communities throughout the Indo-West Pacific. Study of their ecology is, however, hindered by incomplete knowledge of their taxonomy: most species cannot be identified in the field and the two genera cannot always be distinguished reliably. We used a 735-bp fragment of the octocoral-specific mitochondrial protein-coding gene msh1 to construct a phylogeny for 92 specimens identified to 19 species of Lobophytum and 16 species of Sarcophyton . All phylogenetic methods used recovered a tree with three strongly supported clades. One clade included only morphologically typical Sarcophyton species with a stalk distinct from the polypary, poorly formed club-shaped sclerites in the colony surface, and large spindles in the interior of the stalk. A second clade included only morphologically typical Lobophytum colonies with lobes and ridges on the colony surface, poorly formed clubs in the colony surface, and interior sclerites consisting of oval forms with regular girdles of ornamental warts. The third distinct clade included a mix of Sarcophyton and Lobophytum nominal species with intermediate morphologies. Most of the species in this mixed clade had a polypary that was not distinct from the stalk, and the sclerites in the colony surface were clubs with well-defined heads. Within the Sarcophyton clade, specimens identified as Sarcophyton glaucum belonged to six very distinct genetic sub-clades, suggesting that this morphologically heterogeneous species is actually a cryptic species complex. Our results highlight the need for a complete taxonomic revision of these genera, using molecular data to help confirm species boundaries as well as to guide higher taxonomic decisions.