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.     

Acute arthritis of cats associated with feline calicivirus infection

Twelve specific pathogen-free cats were infected either by intra-articular inoculation or by contact exposure to one of two strains of feline calicivirus (FCV), either F65, a field strain originating from an outbreak of lameness in a group of cats, or a vaccine strain. Following either route of exposure, both strains induced signs typical of FCV infection including oral and nasal ulceration, conjunctivitis and ocular discharge. These signs were of equal severity for both virus strains, but overall, following either route of infection, F65 induced more severe disease than the vaccine strain, with marked pyrexia, lethargy and lameness. Vaccine virus only induced a relatively mild lameness following intra-articular inoculation. Gross pathological and histopathological lesions were seen in some of the joints, but again changes were more severe in the F65-exposed cats. Virus was isolated from both normal and affected joints from both groups of F65-exposed cats, and from a joint from each cat inoculated intra-articularly with vaccine virus. Mild transient lameness was also seen in one of two control cats inoculated intra-articularly, but no pathological changes were seen or virus isolated from joints. A cDNA probe used in RNA dot blot hybridisation experiments was found to be specific and more sensitive than virus isolation in detecting FCV in selected tissues. This may be useful in future studies on the pathogenesis of FCV disease and in studies on viral persistence in FCV carriers.     

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     

The electron microscopy of post-growth induced defects of ZnSe/GaAs epilayers

A fundamental research of structural defects induced upon post-growth processing of ZnSe/GaAs epilayers grown on (100) GaAs was done by identifying defect-related reflections in the transmission electron diffraction (TED) patterns of ZnSe. Structural artifacts, other than the as-grown defects, on this material system could be excluded according to our results. Four types of abnormal reflections have been observed in addition to primary reflections. These extra reflections are sensitive to the post-growth processing of ZnSe epilayers and may arise from various external effects, rather than epitaxy growth, such as irradiation damage, surface oxidation, and surface contamination. By mapping these reflections at several major zone axes using TED patterns, we found that the reciprocal lattice for a ZnSe crystal with structural defects consists of two distinct types of extra reflections associated with irradiation damage. The first type of extra reflections is ±1/3{111} and the other is ±1/2{111} corresponding to pure-edge and non-edge dislocation loops, respectively. For (100) oriented wafers, the ±1/3{111} and ±1/2{111} reflections were observed only on two of the four possible 〈111〉 variants (i.e. [111]_Zn and [111]_Zn)and this phenomenon was attributed to the anisotropy of defect distribution. Extra reflections associated with surface oxidation and contamination are also observed. The orientation relationships between a surface hexagonal ZnO and a cubic ZnSe film are [0001]_ZnO//[−111]_ZnSe, and [01−11]_ZnO//[011]_ZnSe. The origin, characterization, and elimination of these induced reflections are discussed. With the knowledge about these extra effects on structural defect formation, we have shown the real microstructure of ZnSe epilayers.     

Evaluation of the aortic root by MRI: insights from patients with homozygous familial hypercholesterolemia

Hepatitis C chronically infects approximately 1.5% of Americans and is the most common clinical problem facing hepatologists. Since the virus was initially described in 1989, development of an effective therapy has been challenging. Although several different therapeutic agents have been used, no therapy has been shown to reliably eradicate the virus. Interferon-alpha, a cytokine with immunostimulatory and anti-viral properties, has become the therapy of choice for patients with chronic hepatitis C infection. Trials assessing the efficacy of interferon-alpha have characterized host and viral factors predictive of responses to treatment. A thorough understanding of these predictive factors is requisite to providing cost-effective therapeutic decisions for the patient with chronic hepatitis C infection.     

SPECT, CT, and MRI in head injury: acute abnormalities followed up at six months

We report a case of cervicofacial necrotizing fasciitis that developed after blepharoplasty, an occurrence that, to our knowledge, has not previously been reported in the medical literature. A patient who presented to our institution 3 days after undergoing blepharoplasty of the upper eyelid was diagnosed as having fulminant fasciitis involving extensive areas of the face, scalp, and neck. We review the case in detail and discuss clinical and radiological clues to diagnosis, surgical and medical management, wound care, and subsequent scar contracture. This case emphasizes the need for individualized, appropriate postoperative care and for an awareness of this rare, potentially fatal complication. Early recognition and aggressive treatment of cervicofacial fasciitis can arrest its rapid progression and prevent devastating sequelae.