Differential methylation of a CpG-island concatemer in hemi- and homozygous transgenic mice.

Methylation-free islands (MFIs), clusters of non-methylated CpG-dinucleotides in mammalian genomes, are associated with a majority of studied genes. By which precise mechanism they maintain their unmethylated status is unknown. The behaviour of transgenic MFIs may contribute to unveil this enigma. We have generated a high-copy number transgenic line with the MFI from the murine Thy-1.1 allele. A stable, minor fraction of this otherwise non-methylated DNA became completely methylated in all adult tissues tested. Furthermore, individuals homozygous for the transgene showed a significantly higher proportion of methylated copies compared to the hemizygous state. These findings support the hypothesis that a limited pool of trans-acting factors are involved in maintaining the hypomethylated state.     

Stereological estimates of nuclear volume correlated with histopathological grading and prognosis of bladder tumour

The aim of this retrospective study is to provide morphometric data which make grading of urinary bladder tumours objective and reproducible by stereological estimation of nuclear volume using the principle of estimating of the volume of particles of arbitrary shape. The study includes 92 specimens: 12 from normal bladder mucosa, and 80 from bladder tumours (15 grade I, 45 grade II, 19 grade III and one grade IV according to Bergkvist et al. 1965). After standard fixation, embedding, sectioning and hematoxylin-eosin staining, an unbiased estimate of the mean volume of nuclei sampled with a change proportional to the volume: (Formula: see text) was calculated. Here l0 is the length of the intercept through a test point hitting a nucleus measured in a random direction through the test point. The weighted means of nuclear volume in bladder tumours are spread over a wide range and show a strong correlation with the Bergkvist grade. Moreover, the relationship between the weighted mean volume of nuclei in bladder tumours and the prognosis is very good. Only one of 35 patients with a mean nuclear volume below 300 micron 3 died of bladder cancer whereas 18 of 19 patients with a mean nuclear volume above 500 micron 3 developed invasives tumours or died of their disease. This simple and fast estimate of nuclear volume seems to provide objective data of high prognostic value.     

The hunting habitats of Goshawks Accipiter gentilis in boreal forests of central Sweden

Radio-tracking was used to study the habitat utilization of hunting Goshawks Accipiter gentilis in central Swedish boreal forest. Data were collected during September-June, in 1977–1981. There was no major difference between autumn, winter or spring in any aspect of habitat preference, despite seasonal differences in prey composition. Despite the strong reversed sexual size-dimorphism, no habitat segregation between the sexes was found. It is argued that this is due to the lack of prey segregation between the sexes. Goshawks showed a strong preference for mature forest, which was chosen twice as often as would be expected from its prevalence in the environment, whereas the younger successional stages of the forest were under-used. Goshawks preferred large habitat patches, and showed no major preference with respect to tree species composition of the forest. Kills made by radio-tagged Goshawks showed the same habitat- and patch-size distribution as did Goshawk locations in general. It is argued that the profitability of different hunting habitats, and thus habitat preferences, is not only determined by prey density in the habitats, but also by other habitat features influencing the Goshawk's ability to hunt there. Winter home-ranges averaged 5700 ha, considerably larger than Goshawk ranges recorded in areas with richer prey supply.     

A new monostiliferous hoplonemertean (Nemertea), Oerstedia striata sp.n., from the west coast of Sweden

Oerstedia striata sp.n. (family Prosorhochmidae?) is described. The species was originally reported as one of the varieties of the polymorphic species Oerstedia dorsalis (Abildgaard, 1806). Its specific status was detected by an electrophoretic screening of six enzymes, of which four were unique for this species as compared to the analysed forms of O. dorsalis.     

A study of the diversity and geographical variation in numbers of leg‐bearing segments in centipedes (Chilopoda: Geophilomorpha) in north‐western Europe

Five species of geophilomorphs, Strigamia maritima (Leach, 1817), Geophilus flavus (De Geer, 1778), Geophilus truncorum Bergsøe and Meinert, 1866, Geophilus proximus C. L. Koch, 1847, and Pachymerium ferrugineum (C. L. Koch, 1835), from various sample sites in north‐western Europe were examined for numbers of leg‐bearing segments. Where data was adequate, mean segment numbers were correlated with latitudinal gradients for each species. Solely in S. maritima and some populations of P. ferrugineum did the results show a highly significant geographic pattern towards more leg‐bearing segments in southern populations of both sexes. As concerns G. proximus, we presented for the first time a remarkable geographic shift towards an increased number of pairs of legs in northern populations. Contrary to the conventional geographic pattern, we found that G. flavus and G. truncorum did not exhibit a north–south increase or decrease in segment number. Although there was no general/universal evidence supporting the occurrence of a significant regression slope between mean segment number and latitude/temperature, more information shows that the overall region‐to‐region segment variation was highly significant in both sexes. In S. maritima and P. ferrugineum mean segment number was correlated with the north–south temperature cline. The same was not observed in G. proximus. Parthenogenesis in G. proximus, and a series of ecological characteristics such as habitat preferences, spatial distribution, and fragmented populations could explain the presence or absence of a geographic patterning of segment number variation along a latitudinal cline. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 100 , 899–909.     

On the orientation of foreign neurotransmitter receptors in Xenopus oocytes

Xenopus oocytes can be made to incorporate into their membrane foreign neurotransmitter receptors and voltage-activated sodium channels. In their original location the receptors are normally activated by the extracellular action of transmitter substances. Tests were made to see if some of the newly synthesized foreign receptors were inserted in the oocyte membrane with their active site facing inwards. Since intracellular injections of acetylcholine, gamma-aminobutyric acid, serotonin and kainic acid and tetrodotoxin into the oocyte failed to elicit a response, we conclude that very few, or none, of the receptor molecules expressed in the oocyte by the exogenous mRNA are inserted with the wrong orientation in the membrane.     

Moving Cell Boundaries Drive Nuclear Shaping during Cell Spreading

The nucleus has a smooth, regular appearance in normal cells, and its shape is greatly altered in human pathologies. Yet, how the cell establishes nuclear shape is not well understood. We imaged the dynamics of nuclear shaping in NIH3T3 fibroblasts. Nuclei translated toward the substratum and began flattening during the early stages of cell spreading. Initially, nuclear height and width correlated with the degree of cell spreading, but over time, reached steady-state values even as the cell continued to spread. Actomyosin activity, actomyosin bundles, microtubules, and intermediate filaments, as well as the LINC complex, were all dispensable for nuclear flattening as long as the cell could spread. Inhibition of actin polymerization as well as myosin light chain kinase with the drug ML7 limited both the initial spreading of cells and flattening of nuclei, and for well-spread cells, inhibition of myosin-II ATPase with the drug blebbistatin decreased cell spreading with associated nuclear rounding. Together, these results show that cell spreading is necessary and sufficient to drive nuclear flattening under a wide range of conditions, including in the presence or absence of myosin activity. To explain this observation, we propose a computational model for nuclear and cell mechanics that shows how frictional transmission of stress from the moving cell boundaries to the nuclear surface shapes the nucleus during early cell spreading. Our results point to a surprisingly simple mechanical system in cells for establishing nuclear shapes.     

Further Studies of Dopamine Metabolism and Function in Tetrahymena1

The large amounts of dopamine accumulated by cells of Tetrahymena pyriformis strain NT-1 and secreted into their growth medium were found to depend primarily upon an extracellular, non-enzymatic conversion of tyrosine to L-dihydroxyphenylalanine (L-DOPA); L-DOPA was then rapidly taken into the ceils and transformed into dopamine enzymatically. Efforts to find physiologically significant dopamine binding sites on the cell surface or dopamine-sensitive adenylate cyclase activity were unsuccessful, suggesting that the catecholamine does not function in Tetrahymena as it does in higher animals.     

Uptake of Sulfate by Roots and Leaf Slices of Barley: Mediated by Single, Multiphasic Mechanisms

The uptake of 10^?1M–2.5 × 10^?1M sulfate by roots and leaf slices of barley can be described by a single isotherm having, respectively, 8 and 5 phases with regularly increasing kinetic constants. Each phase covers a limited concentration range and obeys Michaelis-Menten kinetics. The uptake of sulfate is proposed to be rate-limited by a single mechanism or structure which is located in the plasmalemma (or cytoplasm) and which changes characteristics at certain discrete external concentrations (inflection points). Examination of published data indicates that the uptake of other inorganic ions by higher plant cells is also mediated by single, multi-phasic mechanisms.