The ability of isolated myocardial cells of the rat to contract at temperatures near freezing point

1. 1.|Single spontaneously-beating myocardial cells were prepared by enzymatic isolation from rat heart ventricles. Using a TV camera and videotape recorder, beating parameters were characterized with respect to temperature.

2. 2.|The cells obtained showed two types of contractions: (a) action-potential-coupled, fast contractions, which could be triggered electrically but sometimes also occurred spontaneously; and (b) a phasic type of contraction which occurs spontaneously as a slowly (25–200 μm s⁻¹) proceedings wave through the cell and is not coupled to the action potential.

3. 3.|Both the spontaneous phasic and electrically-triggered fast contractions continued down to about 0°C, where the cells went into an irreversible contracture, but recovered if the temperature was raised before the contracture occurred.

4. 4.|In physically-contracting cells, the beat rate and the velocity of the contraction wave were temperature dependent showing a linear relationship on an Arrhenius plot; apparent activation energies were 31 and 54 kJ mol⁻¹, respectively.

5. 5.|When fast contractions were triggered by electric field stimulation, the threshold of cells was reduced as temperature rose. The rheobase values showed that cell excitability decreased more steeply when the temperature fell below 20°C. This change could not be seen in the values of chronaxie, which showed a linear relationship on the Arrhenius plot within the whole temperature range used (6–35°C).

6. 6.|It is concluded that the lower temperature limit is not the same for all structural levels of the rat heart. Myocardial cells are able to contract still at the freezing point while mechanisms initiating the heart beat require higher temperatures to be functional.

Paleolimnological reflections of fiber-plant retting in the sediment of a small clear-water lake

Likolampi is a small groundwater kettle-hole lake in Ilomantsi, Eastern Finland. At 11 m water depth the uppermost 51 cm of sediment contains about 390 varves. A thin layer of chrysophyte statospores is the main structural unit in these; in some varves it is followed by a layer of cf. Drepanocladus fluitans moss spores. Below the laminated sequence, two layers of moss detritus (110–87 cm, 81–51 cm) alternate with greyish brown fine detritus. Pollen analysis reveals that the start of the deposition of varved sediment coincides with the beginning of intensive fiber plant retting with large quantities of Cannabis/Humulus -type pollen and even regular occurrence of the insect-pollinated Linum usitatissimum. The cf. Drepanocladus spores are almost absent during the retting period (c. 1590–1900 by the varves), but very common before and after it. Drepanocladus fluitans is capable of assimilating dissolved CO₂ in water, so it thrives at low pH and is typical for oligotrophic clear-water lakes. We conclude that the stratigraphy of Drepanocladus fragments and spores is a true negative indicator of cultural influence in Likolampi.     

Prostacyclin,thromboxane and prostaglandin F2αin suction blister fluid of human skin: Effect of systemic aspirin and local glucocorticosteroid treatment

Changes in prostaglandin (PG) metabolism are known to be involved in various skin diseases. To elucidate the behavior of hree vasoactive PGs in human skin, namely prostacyclin (PG1₂), thromboxane A₂ (TxA₂) and PGF_2α, their stable metabolites, 6-keto-PGF_1α. TxB₂, and 13, 14 dihydro-15-keto PGF_2α (MPGF_2α), respectively, were measured by radioimmunoassays in suction blister fluids of 29 healthy male subjects. Nine of them were treated with acetylsalicylic acid (0.5 g × 4/day for one day beforehand), eight with local glucocorticoid (clobetasol-17-propionate, Dermovat^R Cream, twice a day for seven days) and 12 served as controls. All three PGs were detected in blister fluid. In controls the mean (±SD) concentration of 6-keto-PGF_1α was 1160 ± 470 pg/ml (n=12) that of TxB₂ 1590 ± 610 pg/ml (n=12) and that of MPGF_2α 1800 ± 710 pg/ml (n=12), levels which are higher than the respective concentrations in human plasma. The preceding aspirin treatment decreased the 6-keto-PGF_1α levels by 40 % (P<0.005), the TxB₂ levels by 80 % (P<0.001) and MPGF_2α levels by 35 % (P<0.05), whereas the preceding local glucocorticoid caused no changes in these PG levels. The results show that 1) PG1₂, TxA₂, and PGF_2α are locally released in the suction blister fluid of healthy human skin, 2) systemic treatment with a PG synthesis inhibiting drug, acetysalicylic acid, reduces this release, and 3) locally applied clobetasol-17-propionate does not affect the levels of prostaglandins and thromboxane as measured by our methods.     

Electron microscopy of the paracervical (Frankenhäuser) ganglion of the adult rat

Summary The ultrastructure of the paracervical (Frankenhäuser) ganglion in the rat was studied after immersion or perfusion fixation with glutaraldehyde followed by post-osmification. This ganglion is located at the uterovaginal junction in the vicinity of arteria uterina and contains three neuronal cell types. (1) Principal neurons have a fine structure mainly similar to the ganglion cells of other autonomic ganglia. (2) Small granule-containing cells occur in clusters often close to fenestrated capillaries. They are divided into two subgroups according to the size of their cytoplasmic granules; those containing only small granulated vesicles of 800 to 1400 Å in diameter and those having also large granulated vesicles of 2000 to 3000 Å in diameter. (3) Vacuolated nerve cells are large cells that resemble the principal neurons in their cytoplasmic components, except that they contain one to ten vacuoles with corpuscles of different size and shape. The possible physiological significance of the small, granule-containing cells in the uterine function is discussed.     

Time-resolved fluoroimmunoassay of plasma and urine O-desmethylangolensin

We present a method for the determination of the phytoestrogen metabolite O-desmethylangolensin (O-DMA) in plasma (serum) and in urine. O-DMA is a metabolite of daidzein, which occurs in soybeans. It has been suggested that isoflavones may afford protection against breast and prostate cancer and therefore, also the metabolites are of interest. The method is based on time-resolved fluoroimmunoassay (TR–FIA) using a europium chelate as a label. After the synthesis of 4′′-O-carboxymethyl-O-DMA, this compound is coupled to bovine serum albumin, and then used as antigen in immunization of rabbits. The tracers with the europium chelate are synthesized using the same 4′′-O-derivative of the -methyldeoxybenzoin. After enzymatic hydrolysis and ether extraction the immunoassay is carried out by time resolved fluoroimmunoassay (TR–FIA). Cross-reactivity was tested with angolensin, dihydrogenistein, dihydrodaidzein, equol, 6′-OH-angolensin, trans-4-OH-equol, 6′-OH-O-DMA, cis-4-OH-equol and 5-OH-equol. The antiserum cross-reacted only with angolensin. This cross-reactivity seems not to influence the results, which were highly specific. Plasma samples are hydrolyzed and extracted. Urine samples are analyzed directly after hydrolysis without extraction. The correlation coefficient between the plasma TR–FIA results and the GC–MS results was high; r value was 0.985. The correlation coefficient between the urine TR–FIA results and the GC–MS results was high over the entire range of concentrations (0–1500 nmol/l); r value was 0.976, but lower in the low concentration range (0–100 nmol/l), i.e. value was 0.631. The intra-assay coefficients of variation (CVs) for plasma O-DMA concentrations and for urine O-DMA concentrations at three different concentrations varied 2.8–7.7 and 3.0–6.0%, respectively and the inter-assay CVs varied 3.8–8.9 and 4.4–6.6%, respectively. The working range of the plasma and urine O-DMA assays was 0.5–512 nmol/l.     

Highlights of large lake research and management in Europe

Lakes in Europe have a bipolar distribution by latitude with higher lake densities in the north (58–65° N) and south (38–48° N). By area, 95% of the large lakes (>100 km²) are located at altitudes lower than 100 m above sea level (ASL) and only 1% lie higher than 1,000 m ASL. Physically large lakes exhibit several similarities to seas and oceans in their thermal structure and circulation dynamics. From the chemical point of view, lakes are important accumulation sites for substances transported from the watershed or built up in the lake itself but they may contribute positively to global greenhouse gas emission. Fauna and flora of ancient large lakes such as the Caspian Sea and Lake Ohrid include large numbers of endemic species, which become endangered if conditions change because of direct human impact, alien species invasions or climate change. Large lakes offer a wide range of ecosystem services to society, the multiple use of which creates multiple pressures on these water bodies such as nutrient load and toxic pollution, modification of hydrology and shore line structure, and shifting of the food web balance by stocking or harvesting various species. Although large lakes are among the best-studied ecosystems in the world, the application to them of environmental regulations such as the European Water Framework Directive is a challenging task and requires that several natural and management aspects specific to these water bodies are adequately considered.     

Parental overwintering history affects the responses of Thlaspi arvense to warming winters in the North

The overwintering conditions of northern plants are expected to change substantially due to global warming. For perennial plants, winter warming may increase the risk of frost damage if the plants start dehardening prematurely. On the other hand, evergreen plants may remain photosynthetically active and thereby benefit from milder winters. The positive and negative effects of mild winters on annual plants remain, however, largely unknown. We postulated that summer annuals may be susceptible to frost damage if the seeds germinate during a mild spell in winter. Winter annuals may utilize a warm period for photosynthesis. These questions were addressed in two consecutive experiments in which pot-grown individuals of Thlaspi arvense that overwintered in the field were exposed to an elevated temperature for 8 days in growth chambers in mid-winter. No premature germination was observed in summer annuals. However, in accordance with our hypothesis, winter annuals started photosynthesising very rapidly upon exposure to elevated temperature. The winter warming treatment affected neither the total number of seeds produced nor the mean seed weight. These seeds, possessing divergent parental overwintering histories, were used as starting material for the second experiment. Seeds originating from both summer and winter annual plants germinated both in the autumn and in the following spring. We observed a major parental effect associated with the winter warming treatment. The warm spell experienced by the mother plant (either as a winter annual rosette or as a summer annual seed) reduced the proportion of autumn germination in the next generation. Only 43% of the seeds of summer annuals possessing a parental warming history germinated before the winter, whereas the germination percentage of seeds with no previous winter warming history was 71%. In the case of seeds collected from winter annual plants, 4% of the seeds germinated in autumn if the mother plants experienced the warming treatment during the previous winter, whereas the corresponding value was 37% if the mother plants did not experience warming. Our results show that summer and winter annual individuals show diverse responses to warm spells in winter. Since the responses are not limited only to the generation that actually experiences the warm spell, but also appear in their offspring, long-term studies consisting of several generations are called for.     

Determinants of parasitoid communities of willow‐galling sawflies: habitat overrides physiology,host plant and space

Studies on the determinants of plant–herbivore and herbivore–parasitoid associations provide important insights into the origin and maintenance of global and local species richness. If parasitoids are specialists on herbivore niches rather than on herbivore taxa, then alternating escape of herbivores into novel niches and delayed resource tracking by parasitoids could fuel diversification at both trophic levels. We used DNA barcoding to identify parasitoids that attack larvae of seven Pontania sawfly species that induce leaf galls on eight willow species growing in subarctic and arctic–alpine habitats in three geographic locations in northern Fennoscandia, and then applied distance‐ and model‐based multivariate analyses and phylogenetic regression methods to evaluate the hierarchical importance of location, phylogeny and different galler niche dimensions on parasitoid host use. We found statistically significant variation in parasitoid communities across geographic locations and willow host species, but the differences were mainly quantitative due to extensive sharing of enemies among gallers within habitat types. By contrast, the divide between habitats defined two qualitatively different network compartments, because many common parasitoids exhibited strong habitat preference. Galler and parasitoid phylogenies did not explain associations, because distantly related arctic–alpine gallers were attacked by a species‐poor enemy community dominated by two parasitoid species that most likely have independently tracked the gallers’ evolutionary shifts into the novel habitat. Our results indicate that barcode‐ and phylogeny‐based analyses of food webs that span forested vs. tundra or grassland environments could improve our understanding of vertical diversification effects in complex plant–herbivore–parasitoid networks.     

Enemy‐free space and the host range of a lichenivorous moth: a field experiment

According to the enemy‐free space hypothesis (EFS), parasites and predators create a selective force for a specialization on a host that assures better protection against natural enemies than other potential hosts. Few studies have found support for EFS and none of them have covered the whole larval period in natural conditions. I studied the growth and survival of lichen‐feeding moth larvae on five lichen species with and without their natural enemies in natural conditions covering the whole larval period. All the three following EFS predictions gained support. First, natural enemies caused significant mortality of larvae. Second, when natural enemies were present, larval survival was highest on preferred Ramalina lichens. Third, larvae attained higher mass on non‐preferred Parmelia sulcata than on Ramalina species, indicating fecundity cost to feed on Ramalina species instead of P. sulcata. EFS for C. lichenaria larvae on Ramalina species is likely a consequence of shrubby appearance of Ramalina species which provide better larval protection from predation than other hosts.     

Combined use of hard X-ray phase contrast imaging and X-ray fluorescence microscopy for sub-cellular metal quantification

Hard X-ray fluorescence microscopy and magnified phase contrast imaging are combined to obtain quantitative maps of the projected metal concentration in whole cells. The experiments were performed on freeze dried cells at the nano-imaging station ID22NI of the European Synchrotron Radiation Facility (ESRF). X-ray fluorescence analysis gives the areal mass of most major, minor and trace elements; it is validated using a biological standard of known composition. Quantitative phase contrast imaging provides maps of the projected mass and is validated using calibration samples and through comparison with Atomic Force Microscopy and Scanning Transmission Ion Microscopy. Up to now, absolute quantification at the sub-cellular level was impossible using X-ray fluorescence microscopy but can be reached with the use of the proposed approach.