A base-pairing interaction between U2 and U6 small nuclear RNAs occurs in > 150S complexes in HeLa cell extracts: implications for the spliceosome assembly pathway

In mammalian cells, base pairing between the U2 and U6 small nuclear RNAs is required during pre-RNA splicing. We show by psoralen crosslinking of HeLa nuclear extract that U2.U6 base pairing occurs within abundant ribonucleoprotein complexes that sediment at > 150 S in glycerol gradients. All of the spliceosomal RNAs (U1, U2, U4, U5, and U6) cosediment with these large complexes, suggesting that they may be related to small nuclear RNA-containing structures called speckles/coiled bodies or snurposomes, which have been visualized in mammalian or amphibian nuclei, respectively. In contrast to nuclear extract, S100 extract, which is splicing-defective and lacks the > 150S complexes, does not contain base-paired U2.U6. However, U2.U6 base pairs form in S100 extract that has been made splicing-competent by supplementation with Ser/Arg-rich (SR) proteins, ATP, and an adenovirus splicing substrate. During splicing in supplemented S100 extract, U2.U6 base pairing precedes the appearance of splicing intermediates and occurs initially in an approximately 60S spliceosome complex but also in progressively larger (100-300 S) complexes. Possible functional relationships between the 60S spliceosome and the > 150S complexes are discussed.     

A set of electrophoretic molecular markers for strain typing and population genetic studies of Histoplasma capsulatum

A set of eleven biallelic and three multiallelic molecular markers have been developed to analyze populations of Histoplasma capsulatum. All markers are amplified by polymerase chain reaction (PCR) and can be readily scored using minimal amounts of template DNA. The 11 biallelic loci have polymorphic restriction endonuclease sites or small insertions or deletions which may be assessed by agarose gel electrophoresis. These markers are inherited in an unambiguous manner and are ideal for assessing structure and gene flow within US populations of H. capsulatum, but are monomorphic in non-US populations. Both length and sequence variation are present in the multiallelic loci, which can be scored by direct sequencing, polyacrylamide gel electrophoresis, or single-strand conformation polymorphism (SSCP): As they are hypervariable, the multiallelic loci can be used to type isolates and to assess the level of genetic variation within populations. Preliminary results indicate that the three multiallelic markers presented are sufficient to distinguish isolates at the individual level and are polymorphic in both US and non-US populations. This collection of molecular markers will be a useful tool in population and epidemiology studies of H. capsulatum.     

Female house crickets, Acheta domesticus, prefer the chirps of large males

This study demonstrates that invertebrate acoustic signals can provide information about male phenotypic attributes, and that females can use this acoustic information in mate choice to select a phenotypically superior mate. I investigated the relationships between a male acoustic sexual signal, the phenotype of the signaller, and the female response to signal variation. I recorded and analysed the calling songs of male house crickets, Acheta domesticus. The analyses showed that chirps convey information about male size. With the exception of amplitude, the mean number of pulses per chirp was the best predictor of male size. I performed a laboratory tape-playback experiment to determine female preference during phonotaxis. Females preferred tapes playing the chirps of large males, specifically chirps with a greater number of pulses per chirp. Selection on the female preference is discussed.Copyright 1997 The Association for the Study of Animal Behaviour     

Comparing two diagnostic tests against the same "gold standard" in the same sample

Methods are proposed for comparing two diagnostic tests for the same data where a threshold for positive for each test is specified. One method contrasts the diagnostic tests' estimated risks. A second method compares the two tests' kappa coefficients. When thresholds for positive test results are specified a priori, maximum likelihood estimators and their asymptotic variances are derived and test statistics are presented for both case-control and naturalistic methods of sampling. The bootstrap is proposed as a method to assess differences in risk estimators when thresholds for positive test results are chosen by scanning the data. Examples are given to illustrate the methods.