Isolation and characterization of the nuclear matrix from the male xenopus laevis following estrogen administration: Kinetics of [3H] uridine incorporation

At various times following estorgen administration, the nuclear matrix was isolated from the liver of male Xenopus laevis by sucrose gradient centrifugation of nuclei treated with a high-salt buffer and DNase I in the presence of a proteolytic inhibitor (PMSC - phenylmethyl sulfonyl chloride). Electron micrographs of the nuclear matrix demonstrate a sponge-like network attached to a well-defined inner envelope with a ribosome-free outer envelope. Chemical analyses show that the HSB-DNase-treated nuclei consist of 16% DNA, 2% RNA, and 82% protein, a composition that is consistent with that of nuclear matrices isolated from other species. The specific activity of the matrix-associated RNA following estrogen treatment appears to be maximally enhanced after 5 h and decreases until approximately 12 h, when the activity begins to increase again.     

Isolation of single atrial and ventricular cells from the human heart.

The single isolated heart cell has recently emerged as a model for the study of the structure and function of cardiac cells. Heart muscle cells of adult animals of various species have been successfully isolated by enzymatic digestion of intact cardiac tissue. In this paper a dissociation method that yields living cells from atrial and ventricular tissue of young and adult humans is detailed. The cells retain the morphologic features of cells in intact cardiac tissue, and they generate action potentials and contractions in response to electrical stimulation. The study of isolated human heart cells should make a valuable contribution to knowledge of the normal and diseased heart.     

Impaired spatial learning in the APPSwe + PSEN1DeltaE9 bigenic mouse model of Alzheimer's disease

Mice co-expressing the Swedish amyloid precursor protein mutation (APP(Swe)) and exon 9 deletion (DeltaE9) of the PSEN1 gene begin to develop amyloid plaques at 6-7 months of age. We demonstrate here a spatial learning deficit in 7-month-old APP(Swe) + PSEN1DeltaE9 bigenic mice using an adaptation of the Barnes maze. Mice were first trained on a cued target followed by a hidden-target condition. Although bigenic mice quickly learned the cued-target version of the task, they were significantly impaired when switched to the hidden-target version. In contrast, a separate group of double-transgenic mice trained first on the spatial hidden-target version of the task were unimpaired relative to wild-type controls. We propose that processes such as general rule learning, context learning and exploratory habituation exert a greater influence when the testing environment is novel and overshadow the spatial memory deficit in naive bigenic mice. However, when cued-target training is conducted first, these processes habituate and the spatial learning deficit is unmasked. Seven-month-old APP(Swe) + PSEN1DeltaE9 mice were unimpaired on tests of memory that did not involve learning the rules governing spatial associations.     

A winged helix domain in human MUS81 binds DNA and modulates the endonuclease activity of MUS81 complexes

The MUS81-EME1 endonuclease maintains metazoan genomic integrity by cleaving branched DNA structures that arise during the resolution of recombination intermediates. In humans, MUS81 also forms a poorly characterized complex with EME2. Here, we identify and determine the structure of a winged helix (WH) domain from human MUS81, which binds DNA. WH domain mutations greatly reduce binding of the isolated domain to DNA and impact on incision activity of MUS81-EME1/EME2 complexes. Deletion of the WH domain reduces the endonuclease activity of both MUS81-EME1 and MUS81-EME2 complexes, and incisions made by MUS81-EME2 are made closer to the junction on substrates containing a downstream duplex, such as fork structures and nicked Holliday junctions. WH domain mutation or deletion in Schizosaccharomyces pombe phenocopies the DNA-damage sensitivity of strains deleted for mus81. Our results indicate an important role for the WH domain in both yeast and human MUS81 complexes.     

Development of an image-based technique to examine joint congruency at the elbow

Identifying joint contact in articular joints is important for both the biomechanical investigation of joint mechanics and the study of osteoarthritis. The purpose of this study is to develop a proximity mapping technique to non-invasively determine joint congruency, as a surrogate of joint contact. To illustrate the capabilities of this algorithm, a cadaveric upper extremity was positioned at varying degrees of elbow flexion. This technique was validated using a gold standard experimental casting technique. The pattern of the cast showed an excellent agreement with the generated proximity map using the inter-bone distance algorithm. The results from this study agree with the results of previous studies examining joint contact at the elbow both in the location and in the tracking of the joint contact throughout elbow flexion. Ultimately, this technique will lead to an increased understanding of the effect of malalignment and instability of the joint on contact mechanics.     

Detecting detectability: identifying and correcting bias in binary wildlife surveys demonstrates their potential impact on conservation assessments

The European Commission Habitats Directive requires that changes in the conservation status of designated species are monitored. Nocturnal and elusive species are difficult to count directly and thus population trajectories are inferred by variation in the incidence of field signs. Presence/absence techniques are, however, vulnerable to Type II errors (false negatives). The Eurasian otter (Lutra lutra), listed by the IUCN as ‘near threatened’, is monitored throughout Europe using the ‘Standard Otter Survey’ method. We explored the reliability of this approach by analysing species incidence at 1,229 sites throughout Ireland. Naïve species incidence was 72 % [95 % confidence interval (CI), 69–75 %] with variation affected significantly by survey team and, at running freshwater sites, the number of bridges present and rainfall during the month, and most notably during the 7 days, prior to survey. Rainfall had no effect on static freshwater sites or the coast. Marginal estimated mean species incidence derived from a GLM assuming the β coefficient of the survey team associated with the highest prevalence, no rainfall in the week prior to survey and sites that had multiple bridges, was 94 % [95 %CI 78–97 %]. We demonstrate that bias and error in binary wildlife surveys can have a major impact on a conservation assessment even when conducted on an apparently well-known species in a developed country with good infrastructure and a long history of similar ecological studies. Our results provide empirical evidence for further criticisms of the Standard Otter Survey method calling into question its value in monitoring changes in otter populations throughout Europe.     

Mechanisms of Mate Investment in the Polygamous Fowl,Gallus gallus

Male fowl (Gallus gallus) that have recently mated invest in their mates by producing antipredator alarm signals at a higher rate. It remains unclear, however, whether these males are investing judiciously in their mates, or responding more generally to recent mating success. Here, we manipulated each male’s mating experience with two different females to test whether males invest selectively in their mates. For 1 wk, males could interact with both females, but could mate with only one of them. In the second week, we removed either the mated or the unmated female and measured the male’s rate of alarm calling. Males did not invest preferentially in their mates, suggesting that increased alarm calling is a more general response to recent mating experience. This relationship could be based on a relatively simple cognitive rule of thumb or on an underlying physiological mechanism. Testosterone and corticosterone are associated with reproduction and antipredator behaviour in other species and so could provide the necessary physiological link in fowl. To test this, we measured plasma levels of testosterone and corticosterone before, during and after mating. Results show that hormone levels did not change as a function of male mating status and hence cannot provide the link between mating and calling behaviour. Instead, we suggest that a general cognitive mechanism is more likely to explain prudent mate investment in this species.     

Colonic impaction in guinea pigs fed a chemically defined diet: a case report

Pathogen-free female Hartley guinea pigs were divided into four diet groups to determine optimal nitrogen requirements for growth, pregnancy and lactation. Three chemically defined test diets varying in total nitrogen were compared with commercial guinea pig feed. Within 13 days, 24 of 78 animals from the three test diet groups died or required euthanasia. Affected guinea pigs were depressed, lost weight and had distended abdomens. Distension, impaction and, in most cases, rupture of the spiral of the ascending colon and subsequent peritonitis were observed at necropsy. Husbandry changes which resulted in a decreased incidence of impactions included increased acclimatization time on commercial feed post-shipping, decreased animal density per cage and cessation of water medication with oxytetracycline.