Trinucleotide microsatellite loci for the peppered moth (Biston betularia)

Fourteen polymorphic trinucleotide microsatellites were isolated in the peppered moth, Biston betularia, using an enrichment protocol. Moderate to high allelic diversity (three to 22 alleles per locus) was found in a sample from northern England. We are currently using these markers to study the population structure of B. betularia in northern England, where a cline for melanism still exists.     

Influence of repeated rectal ultrasound examinations on hormone profiles and behaviour around oestrus and ovulation in dairy cattle

Frequent rectal ultrasound is often used to assess time of ovulation. This study investigated whether frequent rectal ultrasound examination, affects behavioural oestrus and peri-ovulatory hormone profiles (LH, oestradiol and progesterone). Additionally, the relation between peri-ovulatory hormone profiles, oestrous behaviour and time of ovulation was studied. Oestrus was synchronised in two consecutive cycles of Holstein Friesian cattle (parity from 1 to 6; n = 24 cycles). In 12 of these cycles, time of ovulation was assessed by three-hourly rectal ultrasound (assessment of ovulation time with ultrasound group: UG) the other half served as controls (n = 12; no assessment of ovulation time group: CG). There were no significant differences between the onset of oestrus (33.8 +/- 1.6 h), duration of oestrus (13.4 +/- 0.9 h) or intensity of oestrous behaviour (1047 +/- 180 points) between UG and CG treated animals. Furthermore, LH, oestradiol and progesterone profiles were similar between UG and CG. For UG, ovulation took place 30.2 +/- 1.9 h after onset of oestrus. This interval had the largest variation (21 h) of all parameters studied, ranging between 19 and 40 h after onset of oestrus. The smallest variation (6 h) was found in the timing of ovulation in relation to the LH-peak; ovulation took place 25.3 +/- 0.6 h (range: 21.5-27.5 h) after the peak in LH. This study demonstrated that repeated rectal ultrasound does not alter behavioural oestrus or peri-ovulatory hormone profiles and is therefore a useful tool for assessing time of ovulation. Further research, using ultrasound, can now be carried out to find predictors for time of ovulation.     

Mouse muscle phosphofructokinase is partially phosphorylated.

Phosphofructokinase isolated from mouse skeletal muscle 18 hours after intraperitoneal injection of [${}^{32}\text{P]-}\text{PO}{}_4{}^{\mathrm{3?}}$ contained 0.12 to 0.15 moles of covalently bound phosphate per protomer on the basis of the specific activity of radiolabel in the γ-position of ATP. Under identical conditions, muscle pyruvate kinase and aldolase had no covalently bound phosphate.     

A pilot study of rivastigmine in the treatment of delirium after stroke: A safe alternative

Background

Delirium is a common disorder in the early phase of stroke. Given the presumed cholinergic deficiency in delirium, we tested treatment with the acetylcholinesterase inhibitor rivastigmine.

Methods

This pilot study was performed within an epidemiological study. In 527 consecutive stroke patients presence of delirium was assessed during the first week with the confusion assessment method. Severity was scored with the delirium rating scale (DRS). Sixty-two patients developed a delirium in the acute phase of stroke. Only patients with a severe and persistent delirium (defined as a DRS of 12 or more for more than 24 hours) were enrolled in the present study. In total 26 fulfilled these criteria of whom 17 were treated with orally administered rivastigmine with a total dose between 3 and 12 mg a day. Eight patients could not be treated because of dysphagia and one because of early discharge.

Results

No major side effects were recorded. In 16 patients there was a considerable decrease in severity of delirium. The mean DRS declined from 14.8 on day one to 8.5 after therapy and 5.6 after tapering. The mean duration of delirium was 6.7 days (range; 2–17).

Conclusion

Rivastigmine is safe in stroke patients with delirium even after rapid titration. In the majority of patients the delirium improved after treatment. A randomized controlled trial is needed to establish the usefulness of rivastigmine in delirium after stroke.

Trial registration

Nederlands Trial Register NTR1395

     

Sexual selection constrained by life history in a butterfly

Trait evolution via sexual selection has traditionally been viewed as isolated from life-history constraints. Recent theoretical treatments, however, predict that costly sexually selected characters should be mediated by the same allocational trade-offs that apply to more conventional aspects of reproductive investment. Participation in risky competitive behaviours, for example male-male combat, should therefore increase as the opportunity for future reproduction declines. However, the demonstration of such trade-offs has proven to be elusive due to concomitant age-based variation in the physical determinants of fighting 'ability'. Here, I exploit the unique nature of a butterfly contest system to provide compelling evidence for lifetime partitioning of risky and aggressive sexually selected behaviours. I show that male Hypolimnas bolina become more willing to persist in contests over mating territories, and more generally accepting of injury risks, as they age. Contest persistence in this species is not mediated simply by physical condition, and I experimentally isolate the effect of ageing per se from resource ownership and previous contest experience. These results demonstrate how sexually selected behaviours can be ultimately mediated by a shifting trade-off between contemporary reproductive effort and future opportunities.     

CORTICAL CHANGES IN GROWING OOCYTES AND IN FERTILIZED OR PRICKED EGGS OF RANA PIPIENS

The folded cortex of the growing oocyte of the frog extends as microvilli into the substance of the developing vitelline membrane and, internal to the folds, possesses a layer of cortical granules. Free ribosomes, smooth-walled vesicles, coated vesicles, tubules, and electron-opaque granules are abundant in the peripheral zone of the cortex. Mitochondria, lipochondria, pigment granules, and electron-opaque granules are conspicuous between cortical granules and in the underlying endoplasm. Yolk platelets are restricted to the endoplasm. Cortical granules contain neutral and acid mucopolysaccharides, and possibly protein. In the mature oocyte, microvilli are withdrawn and the surface folds eliminated. Cortical granules now lie close to the plasma membrane, sometimes contacting it. Fertilization or pricking causes a wave of breakdown of cortical granules lasting 1–1½ min. Breakdown begins immediately after pricking but not until about 10–15 min after insemination, because the fertilizing sperm takes that long to penetrate the jelly and vitelline membrane. Cortical granules erupt through the surface and discharge their contents into the perivitelline space. Cortical craters left at sites of eruption soon disappear, and pseudopodial protrusions retract. By 30 min after insemination, the surface of the egg is relatively smooth.     

Substrate specificity of a multifunctional calmodulin-dependent protein kinase

The substrate specificity of the multifunctional calmodulin-dependent protein kinase from skeletal muscle has been studied using a series of synthetic peptide analogs. The enzyme phosphorylated a synthetic peptide corresponding to the NH2-terminal 10 residues of glycogen synthase, Pro-Leu-Ser-Arg-Thr-Leu-Ser-Val-Ser-Ser-NH2, stoichiometrically at Ser-7, the same residue phosphorylated in the parent protein. The synthetic peptide was phosphorylated with a Vmax of 12.5 mumol X min-1 X mg-1 and an apparent Km of 7.5 microM compared to values of 1.2 mumol X min-1 X mg-1 and 3.1 microM, respectively, for glycogen synthase. Similarly, a synthetic peptide corresponding to the NH2-terminal 23 residues of smooth muscle myosin light chain was readily phosphorylated on Ser-19 with a Km of 4 microM and a Vmax of 5.4 mumol X min-1 X mg-1. The importance of the arginine 3 residues NH2-terminal to the phosphorylated serine in each of these peptides was evident from experiments in which this arginine was substituted by either leucine or alanine, as well as from experiments in which its position in the myosin light chain sequence was varied. Positioning arginine 16 at residues 14 or 17 abolished phosphorylation, while location at residue 15 not only decreased Vmax 14-fold but switched the major site of phosphorylation from Ser-19 to Thr-18. It is concluded that the sequence Arg-X-Y-Ser(Thr) represents the minimum specificity determinant for the multifunctional calmodulin-dependent protein kinases. Studies with various synthetic peptide substrates and their analogs revealed that the specificity determinants of the multifunctional calmodulin-dependent protein kinase were distinct from several other "arginine-requiring" protein kinases.     

Analysis of skeletal muscle function in the C57BL6/SV129 syncoilin knockout mouse

Syncoilin is a 64-kDa intermediate filament protein expressed in skeletal muscle and enriched at the perinucleus, sarcolemma, and myotendinous and neuromuscular junctions. Due to its pattern of cellular localization and binding partners, syncoilin is an ideal candidate to be both an important structural component of myocytes and a potential mediator of inherited myopathies. Here we present a report of a knockout mouse model for syncoilin and the results of an investigation into the effect of a syncoilin null state on striated muscle function in 6-8-week-old mice. An analysis of proteins known to associate with syncoilin showed that ablation of syncoilin had no effect on absolute expression or spatial localization of desmin or alpha dystrobrevin. Our syncoilin-null animal exhibited no differences in cardiotoxin-induced muscle regeneration, voluntary wheel running, or enforced treadmill exercise capacity, relative to wild-type controls. Finally, a mechanical investigation of isolated soleus and extensor digitorum longus indicated a potential differential reduction in muscle strength and resilience. We are the first to present data identifying an increased susceptibility to muscle damage in response to an extended forced exercise regime in syncoilin-deficient muscle. This study establishes a second viable syncoilin knockout model and highlights the importance of further investigations to determine the role of syncoilin in skeletal muscle.     

Protein kinase recognition sequence motifs

Protein kinases play a crucial role in the regulation of many cellular processes. They alter the functions of their target proteins by phosphorylating specific serine, threonine and tyrosine residues. Identification of phosphorylation site sequences and studies with corresponding model peptides have provided clues to how these important enzymes recognize their substrate proteins. This knowledge has made it possible to identify potential sites of phosphorylation in newly sequenced proteins as well as to construct specific model substrates and inhibitors.