Behavioural responses of Mallorcan midwife toad tadpoles to natural and unnatural snake predators

Calibration of phototherapy equipment can prove to be difficult. One problem is that the self-shielding produced by a patient reduces the irradiance relative to that determined when the cabinet is empty. A model has been developed to determine the factor to apply to the irradiance measured with the cabinet empty to give the irradiance with the patient present, i.e. the self-shielding correction factor. The model assumes that the cabinet consists of a number of discrete infinite line sources backed by perfect mirrors. The patient is treated as a barrier that prevents some of these sources being seen by the detector in the mirror it faces. The model was tested using a Waldmann 8001 K unit and three UV meters for UVA and UVB sources. The measurements suggested some modifications to the model--for UVA multiple reflections were important and for UVB the reflectors were only 30% efficient. The correction factors obtained were 0.87 for UVA and 0.96 for UVB.     

The UL8 subunit of the herpes simplex virus type-1 DNA helicase-primase optimizes utilization of DNA templates covered by the homologous single-strand DNA-binding protein ICP8

The herpes simplex virus type-1 DNA helicase-primase is a heterotrimer encoded by the UL5, UL8, and UL52 genes. The core enzyme, specified by the UL5 and UL52 genes, retains DNA helicase, DNA-dependent nucleoside triphosphatase, and primase activities. The UL8 subunit has previously been implicated in increasing primer stability and in stimulating primer synthesis by the core enzyme. To further characterize the function of the UL8 subunit, we have examined its effect on the activities of the UL5/52 core enzyme using DNA templates covered by the herpes simplex virus type-1 single-strand DNA-binding protein ICP8. We found that while ICP8 stimulated the DNA helicase activity of the UL5/52 proteins up to 3-fold, maximum stimulation by ICP8 required the presence of UL8 protein. Moreover, UL8 protein was required to reverse the inhibitory effect of ICP8 on the DNA-dependent ATPase and primase activities of the UL5/52 proteins. These observations were specific for ICP8 since the heterologous Escherichia coli single-strand DNA-binding protein could not substitute for ICP8. These data suggest that UL8 protein mediates an interaction between the UL5/52 core enzyme and ICP8 that optimizes the utilization of ICP8-covered DNA templates during DNA replication.     

Fixation conditions, the foveola and saccadic latency

Saccades are often elicited in the laboratory by the abrupt step-displacement of a single lit point which is initially the foveolar fixation point and then the eccentric refixation target. This was our Control condition. Four experiments modified the fixation arrangements to examine the effect of altered foveolar stimulation on saccadic latency and accuracy to targets within the central +/- 6 deg of the visual field. (1) No foveolar fixation point: The subject fixated the empty space midway between a pair of fixation guides, which later collapsed into a single refixation target. Latencies for small saccades were similar to the Control values. (2) No foveolar fixation point and no real refixation target: A pair of fixation guides underwent a yoked displacement, and it was easy to fixate and track the invisible midpoint. The smallest saccades were hypermetric, and the typical pattern of latency variation with retinal eccentricity was exaggerated in scale. (3) Spatial effects of a persistent non-target: The precise position of a non-target was important, latency increases being in the ipsilateral hemifield when the non-target was intrafoveolar and unilateral, bilateral when intrafoveolar and on the midline, and local when the non-target was extrafoveolar. (4) Temporal effects of a foveolar fixation point: Blanking an otherwise persistent fixation point for as little as 1 msec at the time of target presentation reduced the expected latency increase. We conclude that the position and timing of foveolar illumination can be critical for saccades of all sizes.     

New genetic techniques for group B streptococci: high-efficiency transformation, maintenance of temperature-sensitive pWV01 plasmids, and mutagenesis with Tn917

Three techniques were developed to improve the genetic manipulation of group B streptococci (GBS). We first optimized a protocol for transformation of GBS by electroporation, which provided transformation efficiencies of 10(5) CFU/microgram. Variables that influenced the transformation efficiency were the glycine content of the competent cell growth media, the electric field strength during electroporation, the electroporation buffer composition, the host origin of the transforming plasmid, and the concentration of selective antibiotic at the final plating. Our transformation protocol provides an efficiency sufficient for cloning from ligation reactions directly into GBS, obviating an intermediate host such as Escherichia coli. Second, temperature-sensitive plasmids of the pWV01 lineage were shown to transform GBS, and their temperature-sensitive replication was confirmed. Lastly, the temperature-sensitive pWV01 plasmid pTV1OK, which contains Tn917, was used as a transposon delivery vector for the construction of genomic Tn917 mutant libraries. We have shown, for the first time, that Tn917 transposes to the GBS chromosome and at a frequency of 10(-3)/CFU. Furthermore, representative clones from a Tn917 library contained single transposon insertions that were randomly located throughout the chromosome. These techniques should provide useful methods for cloning, mutagenesis, and characterization of genes from GBS.     

A beta amyloidogenesis: unique, or variation on a systemic theme?

For more than a century amyloid was considered to be an interesting, unique, but inconsequential pathologic entity that rarely caused significant clinical problems. We now recognize that amyloid is not one entity. In vivo it is a uniform organization of a disease, or process, specific protein co-deposited with a set of common structural components. Amyloid has been implicated in the pathogenesis of diseases affecting millions of patients. These range from Alzheimer's disease, adult-onset diabetes, consequences of prolonged renal dialysis, to the historically recognized systemic forms associated with inflammation and plasma cell disturbances. Strong evidence is emerging that even when deposited in local organ sites significant physiologic effects may ensue. With emphasis on A beta amyloid, we review the present definition, classification, and general in vivo pathogenetic events believed to be involved in the deposition of amyloids. This encompasses the need for an adequate amyloid precursor protein pool, whether precursor proteolysis is required prior to deposition, amyloidogenic amino acid sequences, fibrillogenic nucleating particles, and an in vivo microenvironment conducive to fibrillogenesis. The latter includes several components that seem to be part of all amyloids. The role these common components may play in amyloid accumulation, why amyloids tend to be associated with basement membranes, and how one may use these findings for anti-amyloid therapeutic strategies is also examined.     

Detection of Ehrlichia risticii, the agent of Potomac horse fever, in freshwater stream snails (Pleuroceridae: Juga spp.) from northern California

Ehrlichia DNA was identified by nested PCR in operculate snails (Pleuroceridae: Juga spp.) collected from stream water in a northern California pasture in which Potomac horse fever (PHF) is enzootic. Sequencing of PCR-amplified DNA from a suite of genes (the 16S rRNA, groESL heat shock operon, 51-kDa major antigen genes) indicated that the source organism closely resembled Ehrlichia risticii, the causative agent of PHF. The minimum percentage of Juga spp. harboring the organism in the population studied was 3.5% (2 of 57 snails). No ehrlichia DNA was found in tissues of 123 lymnaeid, physid, and planorbid snails collected at the same site. These data suggest that pleurocerid stream snails may play a role in the life cycle of E. risticii in northern California.