Dual immunodetection and hybridisation in situ of opsin mRNA and rhodopsin protein in retinal sections

We describe a simple method for combining in situ hybridisation and immunohistochemistry on the same retinal section. The technique was developed using a radiolabelled cDNA probe for opsin and an antibody (ROS1F4) against rhodopsin. This method retains the antigenic sites if immunocytochemistry is performed prior to in situ hybridisation. Opsin mRNA was found in the photoreceptor inner segment with rhodopsin immunolocalised to the photoreceptor outer segments. The technique should be applicable to numerous situations including analysis of the sequence of events in the expression and synthesis of the various opsins during retinal development and degeneration.     

Fire evacuation skills: Cognitive-behavioral versus computer-mediated instruction

Fifty-six fourth-grade children participated in this study to assess three training procedures for relative effectiveness in acquiring fire emergency skills and attaining knowledge. The first training approach was behavioral, the second used animated graphics, and the third used still graphics. Each of the training groups was compared to a control group that received no treatment. Performance on dependent measures was assessed before and immediately after training. It was hypothesized that the behavioral and animated-graphics conditions would produce the greatest level of skill and knowledge acquisition. Results demonstrated a significantly higher level of fire emergency skill acquisition and knowledge attainment for both the behavioral and animated-graphics groups relative to the still-graphics group and the control group, which had no training. The value of using computer-mediated strategies for fire-safety skills training was demonstrated.     

Genetic fusions of subunit c in the F0 sector of H+-transporting ATP synthase. Functional dimers and trimers and determination of stoichiometry by cross-linking analysis

The multicopy c subunit of the H+-transporting ATP synthase of Escherichia coli folds through the transmembrane F0 sector as a hairpin of two hydrophobic alpha-helices with the proton-translocating aspartyl-61 side chain centered in the second transmembrane helix. The number of subunits c in the F0 complex, which is thought to determine the H+-pumping/ATP stoichiometry, was previously not determined with exactness but thought to range from 9-12. The studies described here indicate that the exact number is 12. Based upon the precedent of the subunit c in vacuolar-type ATPases, which are composed of four transmembrane helices and seem to have evolved by gene duplication of an F0-type progenitor gene, we constructed genetically fused dimers and trimers of E. coli subunit c. Both the dimeric and trimeric forms proved to be functional. These results indicate that the total number of subunit c in F0 should be a multiple of 2 and 3. Based upon a previous study in which the oligomeric organization of c subunits in F0 was determined by cross-linking of Cys-substituted subunits (Jones, P. C. , Jiang, W., and Fillingame, R. H. (1998) J. Biol. Chem. 273, 17178-17185), we introduced Cys into the first and last transmembrane helices of subunit c monomers, dimers, and trimers and attempted to generate cross-linked products by oxidation with Cu(II)-(1,10-phenanthroline)2. Double Cys substitutions at two sets of positions gave rise to extensive cross-linked multimers. Multimers of the monomer that extended up to the position of c12 were correlated and calibrated with distinct cross-linked species of the appropriate doubly Cys-substituted dimers (i.e. c2, c4, . c12) and doubly Cys-substituted trimers (i.e. c3, c6, c9, c12). The results show that there are 12 copies of subunit c per F0 in E. coli, the exact number having both mechanistic and structural significance.     

Ahch, the mouse homologue of DAX1: cloning, characterization and synteny with GyK, the glycerol kinase locus

We cloned the murine full-length cDNA encoding Ahch, the mouse homologue of DAX1 (DSS-AHC Region on Human X Chromosome, Gene1) which is the gene responsible for human X-linked adrenal hypoplasia congenita (AHC) and hypogonadotropic hypogonadism (HH). Sequence analysis revealed that the murine and human cDNAs have 65% aa identity and 75% aa similarity overall. The cysteine residues in the putative DNA binding domain, which may interact with Zn2+ ions to form zinc fingers, are 100% conserved between the two species, indicating that the novel zinc-finger structures in DAX1 may be functional. In addition, mouse interspecific backcrosses show that the Ahch gene is closely linked to the glycerol kinase locus, GyK, on the mouse X chromosome, indicating that the order of the loci is conserved in this syntenic region between mouse and human.     

Equivalence of generalized joint signal representations ofarbitrary variables

Joint signal representations (JSRs) of arbitrary variables generalize time-frequency representations (TFRs) to a much broader class of nonstationary signal characteristics. Two main distributional approaches to JSRs of arbitrary variables have been proposed by Cohen (see Time-Frequency Analysis, Englewood Cliffs, NJ, Prentice Hall, 1995 and Proc. SPIE 1566, San Diego, 1991) and Baraniuk (see Proc. IEEE Int. Conf. Acoust., Speech Signal Processing, ICASSP'94, vol.3, p.357-60, 1994). Cohen's method is a direct extension of his original formulation of TFRs, and Baraniuk's approach is based on a group theoretic formulation; both use the powerful concept of associating variables with operators. One of the main results of the paper is that despite their apparent differences, the two approaches to generalized JSRs are completely equivalent. Remarkably, the JSRs of the two methods are simply related via axis warping transformations, with the broad implication that JSRs with radically different covariance properties can be generated efficiently from JSRs of Cohen's method via simple pre- and post-processing. The development in this paper, which is illustrated with examples, also illuminates other related issues in the theory of generalized JSRs. In particular, we derive an explicit relationship between the Hermitian operators in Cohen's method and the unitary operators in Baraniuk's approach, thereby establishing the relationship between the two types of operator correspondences     

Shear madness: new orthonormal bases and frames using chirpfunctions

The proportional-bandwidth and constant-bandwidth time-frequency signal decompositions of the wavelet, Gabor, and Wilson orthonormal bases have attracted substantial interest for representing nonstationary signals. However, these representations are limited in that they are based on rectangular tessellations of the time-frequency plane. While much effort has gone into methods for designing nice wavelet and window functions for these frameworks, little consideration has been given to methods for constructing orthonormal bases employing nonrectangular time-frequency tilings. The authors take a first step in this direction by deriving two new families of orthonormal bases and frames employing elements that shear, or chirp, in the time-frequency plane, in addition to translate and scale. The new scale-shear fan bases and shift-shear chevron bases are obtained by operating on an existing: wavelet, Gabor (1946), or Wilson basis set with two special unitary warping transformations. In addition to the theoretical benefit of broadening the class of valid time-frequency plane tilings, these new bases could possibly also be useful for representing certain types of signals, such as chirping and dispersed signals