Seed Number, Seed Size and Seed Diversity in Washington Lupin (Lupinus polyphyllus Lindl.)

Over 20 000 seeds of the Washington lupin (Lupinus polyphyllusLindl.) were examined and measured in an experiment carriedout over a 10 year period (1989–1999). Four differentgroups of Washington lupin seeds were found: dark, patterned,grey and light seeds. During the 10 year experiment, the totalaverage number of seeds per plant decreased from 2654 (1990)to 1220 (1999), there was a slight decrease in seed weight perplant and an increase in the average weight per seed. Therewas a clear seed size/number trade-off at the intraspecificlevel. The relative proportions of each seed group also changedwith patterned seeds becoming dominant (50% at the beginningand 90% at the end of the experiment), grey seeds remained constant(constituting 10% of the total seeds) and the proportion ofboth dark (33% at the beginning and 5% at the end) and light(10% at the beginning and 5% at the end of the experiment) seedsdecreased. Six hundred and sixty nine seeds were found to havea different testa ornamentation; they were distributed amongthe different groups as follows: 48% patterned seeds, 29% darkseeds, 12% grey seeds and 11% light seeds. There were no significantdifferences in the physical dimensions of the ornamented seedscompared with all other seeds. The results suggest that theWashington lupin is a mixture of different species and botanicalforms; this is discussed in relation to possible selection pressuresto produce both smaller and larger seeds. The possibility thatchanges in testa ornamentation are influenced by genes controllingthe synthesis of the seed coat pigment is discussed. Copyright2001 Annals of Botany Company Lupinus polyphyllus Lindl., lupine, plant biotechnology, plant parameters, selection pressure, seed size/number trade-off, testa ornamentation     

Two-photon excitation fluorometric measurement of homogeneous microparticle immunoassay for C-reactive protein

Recent developments in infrared laser technology have enabled the design of a compact instrumentation for two-photon excitation microparticle fluorometry (TPX). The microparticles can be used in immunoassays as the antibody-coated solid phase to capture an antigen and then detect it with a fluorescently labeled tracer antibody. Unlike most other methods, TPX technology allows low-volume, homogeneous immunoassays with real-time measurements of assay particles in the presence of a moderate excess of fluorescent tracer. In this study, the TPX assay system was used for the reagent characterization and the measurement of C-reactive protein (CRP) in diluted plasma samples, targeting the assay range useful in infectious disease diagnosis. The pentameric structure of the CRP permitted the optimization of an assay with the lowest detectable concentration of 1 microg/L (7.5 pM) by using a single monoclonal antibody both for capture and as the tracer. With a 1:200 predilution of samples, the measurement range of the assay was 1-150 mg/L, but an additional 1:10 dilution was required for higher concentrations. The TPX method showed a good correlation with the reference result obtained in a routine hospital laboratory, demonstrating the feasibility of the technology for immunodiagnostic applications.     

Targeted disruption of spermidine/spermine N1-acetyltransferase gene in mouse embryonic stem cells. Effects on polyamine homeostasis and sensitivity to polyamine analogues

We have generated mouse embryonic stem cells with targeted disruption of spermidine/spermine N(1)-acetyltransferase (SSAT) gene. The targeted cells did not contain any inducible SSAT activity, and the SSAT protein was not present. The SSAT-deficient cells proliferated normally and appeared to maintain otherwise similar polyamine pools as did the wild-type cells, with the possible exception of constantly elevated (about 30%) cellular spermidine. As expected, the mutated cells were significantly more resistant toward the growth-inhibitory action of polyamine analogues, such as N(1),N(11)-diethylnorspermine. However, this resistance was not directly attributable to cellular depletion of the higher polyamines spermidine and spermine, as the analogue depleted the polyamine pools almost equally effectively in both wild-type and SSAT-deficient cells. Tracer experiments with [C(14)]-labeled spermidine revealed that SSAT activity is essential for the back-conversion of spermidine to putrescine as radioactive N(1)-acetylspermidine and putrescine were readily detectable in N(1),N(11)-diethylnorspermine-exposed wild-type cells but not in SSAT-deficient cells. Similar experiments with [C(14)]spermine indicated that the latter polyamine was converted to spermidine in both cell lines and, unexpectedly, more effectively in the targeted cells than in the parental cells. This back-conversion was only partly inhibited by MDL72527, an inhibitor of polyamine oxidase. These results indicated that SSAT does not play a major role in the maintenance of polyamine homeostasis, and the toxicity exerted by polyamine analogues is largely not based on SSAT-induced depletion of the natural polyamines. Moreover, embryonic stem cells appear to operate an SSAT-independent system for the back-conversion of spermine to spermidine.     

TopBP1 and ATR colocalization at meiotic chromosomes: role of TopBP1/Cut5 in the meiotic recombination checkpoint

Mammalian TopBP1 is a BRCT domain-containing protein whose function in mitotic cells is linked to replication and DNA damage checkpoint. Here, we study its possible role during meiosis in mice. TopBP1 foci are abundant during early prophase I and localize mainly to histone gamma-H2AX-positive domains, where DNA double-strand breaks (required to initiate recombination) occur. Strikingly, TopBP1 showed a pattern almost identical to that of ATR, a PI3K-like kinase involved in mitotic DNA damage checkpoint. In the synapsis-defective Fkbp6(-/-) mouse, TopBP1 heavily stains unsynapsed regions of chromosomes. We also tested whether Schizosaccharomyces pombe Cut5 (the TopBP1 homologue) plays a role in the meiotic recombination checkpoint, like spRad3, the ATR homologue. Indeed, we found that a cut5 mutation suppresses the checkpoint-dependent meiotic delay of a meiotic recombination defective mutant, indicating a direct role of the Cut5 protein in the meiotic checkpoint. Our findings suggest that ATR and TopBP1 monitor meiotic recombination and are required for activation of the meiotic recombination checkpoint.     

Role of adenosine in regulating the heterogeneity of skeletal muscle blood flow during exercise in humans.

Evidence from both animal and human studies suggests that adenosine plays a role in the regulation of exercise hyperemia in skeletal muscle. We tested whether adenosine also plays a role in the regulation of blood flow (BF) distribution and heterogeneity among and within quadriceps femoris (QF) muscles during exercise, measured using positron emission tomography. In six healthy young women, BF was measured at rest and then during three incremental low and moderate intermittent isometric one-legged knee-extension exercise intensities without and with theophylline-induced nonselective adenosine receptor blockade. BF heterogeneity within muscles was calculated from 16-mm(3) voxels in BF images and heterogeneity among the muscles from the mean values of the four QF compartments. Mean BF in the whole QF and its four parts increased, and heterogeneity decreased with workload both without and with theophylline (P < 0.001). Adenosine receptor blockade did not have any effect on mean bulk BF or BF heterogeneity among the QF muscles, yet blockade increased within-muscle BF heterogeneity in all four QF muscles (P = 0.03). Taken together, these results show that BF becomes less heterogeneous with increasing exercise intensity in the QF muscle group. Adenosine seems to play a role in muscle BF heterogeneity even in the absence of changes in bulk BF at low and moderate one-leg intermittent isometric exercise intensities.     

Prevalence of cold-related complaints,symptoms and injuries in the general population: the FINRISK 2002 cold substudy

The prevalence of cold-related complaints and symptoms in the general population has remained unknown. As part of the nationwide FINRISK 2002 health survey performed in Finland, 8,723 people aged 25–64 years filled in a questionnaire asking about the number of hours spent weekly in cold air, their sensations during cold exposure, cold-related complaints, symptoms of diseases, and degradation of performance. Cold thermal sensations at +5°C to −5°C were reported by 35% of men and 46% of women. Almost all subjects reported at least some cold-related complaints, most commonly musculoskeletal pain (men 30%, women 27%), followed by respiratory (25% / 29%), white finger (15% / 18%) and episodic peripheral circulation symptoms (12% / 15%). Decreased mental or physical performance in cold was reported by 75% of men and 70% of women, most commonly impairing manual dexterity and tactile sense. With declining temperature, the first symptom to emerge was pain in the elbow or the forearm (at −3°C), followed by increased excretion of mucus from the lungs (−5°C), while most other symptoms appeared only at lower temperatures of −15°C to −20°C. Most symptoms showed little or no association with the weekly duration of exposure, with the exception of cold-induced pain at most sites. Although, in general, Finns are well adapted to the cold climate, the high prevalence of cold-related complaints poses a challenge to the health care system in terms of decreased performance and the possibility that such symptoms predict more serious health effects, such as increased mortality. This work was carried out in the framework of the FINADAPT project (Assessing the adaptive capacity of the Finnish environment and society under a changing climate).     

Role of hypusinated eukaryotic translation initiation factor 5A in polyamine depletion-induced cytostasis

We have earlier shown that alpha-methylated spermidine and spermine analogues rescue cells from polyamine depletion-induced growth inhibition and maintain pancreatic integrity under severe polyamine deprivation. However, because alpha-methylspermidine can serve as a precursor of hypusine, an integral part of functional eukaryotic translation initiation factor 5A required for cell proliferation, and because alpha, omega-bismethylspermine can be converted to methylspermidine, it is not entirely clear whether the restoration of cell growth is actually attributable to hypusine formed from these polyamine analogues. Here, we have used optically active isomers of methylated spermidine and spermine and show that polyamine depletion-induced acute cytostasis in cultured cells could be reversed by all the isomers of the methylpolyamines irrespective of whether they served or not as precursors of hypusine. In transgenic rats with activated polyamine catabolism, all the isomers similarly restored liver regeneration and reduced plasma alpha-amylase activity associated with induced pancreatitis. Under the above experimental conditions, the (S, S)- but not the (R, R)-isomer of bismethylspermine was converted to methylspermidine apparently through the action of spermine oxidase strongly preferring the (S, S)-isomer. Of the analogues, however, only (S)-methylspermidine sustained cell growth during prolonged (more than 1 week) inhibition of polyamine biosynthesis. It was also the only isomer efficiently converted to hypusine, indicating that deoxyhypusine synthase likewise possesses hidden stereospecificity. Taken together, the results show that growth inhibition in response to polyamine depletion involves two phases, an acute and a late hypusine-dependent phase.     

Involvement of endogenous ouabain-like compound in the cardiac hypertrophic process in vivo

The Na(+)/K(+)-ATPase inhibitor ouabain has been shown to trigger hypertrophic growth of cultured cardiomyocytes; however, the significance of endogenous ouabain-like compound (OLC) in the hypertrophic process in vivo is unknown. Here we characterized the involvement of OLC in left ventricular (LV) hypertrophy induced by norepinephrine (NE) and angiotensin II (Ang II) infusions in rats. Administration of NE (300 microg/kg/h) via subcutanously implanted osmotic minipumps for 72 h resulted in a significant increase in left ventricular weight to body weight (LVW/BW) ratio (P<0.001) and a substantial up-regulation of atrial natriuretic peptide (ANP) gene expression (13.2-fold, P<0.001). NE infusion induced a transient increase in plasma OLC levels at 12 h (P<0.05), which returned to control levels by 72 h. Adrenalectomy markedly reduced both basal and NE-induced increase in plasma OLC levels. LVW/BW ratio was not modulated by adrenalectomy; however, ANP gene expression was blunted by 44% (P<0.01) and 47% (P<0.05) at 12 and 72 h, respectively. In agreement, adrenalectomy reduced up-regulation of ANP without affecting LV mass in rats infused with Ang II (33 microg/kg/h). Administration of exogenous ouabain (1 nM to 100 microM) for 24 h had no effect on ANP gene expression in cultured neonatal rat ventricular myocytes. However, the up-regulation of ANP mRNA levels induced by the alpha-adrenergic agonist phenylephrine (1 microM) was markedly enhanced by ouabain (100 microM) (5.6-fold vs. 9.6-fold, P<0.01). These data show that OLC as an adrenal-derived factor may be required for the induction LV ANP gene expression during the hypertrophic process.     

What do male tench, Tinca tinca, advertise with morphological ornaments?

In contrast to females, tench Tinca tinca (L.) males have large pelvic fins with a thickened and bent second fin ray. Males also produce notable ventral protuberances during breeding, but the function of these male ornaments is not known. Using wild-caught fish, we found that both the size of pelvic fins and ventral protuberance were dependent on body mass/length ratio but not necessarily on condition. Plasma testosterone concentration and relative gonad mass were positively correlated with condition factor. Plasma testosterone concentration was not related to measures of non-specific immune function, but correlated positively with the size of pelvic fins corrected for body length. However, the studied male ornaments were not related to the measures of immune defence or to the load of Diplostomum spp. We did not find evidence for male–male dominance or female preference for large male characteristics within the natural variation in these traits. Thus, our study suggests that also other mechanisms than sexual selection on good genes might contribute to the maintenance of sexual dimorphism in tench.