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.     

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.     

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.     

Library access: success through partnership

A new method of impulse magnetic therapy was studied in the treatment of neurologic complications of osteochondrosis and deforming spondylarthrosis in 225 patients with neurologic manifestations of pathology in cervical or sacral parts of spinal column. To assess the results obtained, EMG, thermography and determination of the levels of blood plasma lipid peroxidation were used.     

Prediction of Fatigue Performance in Aluminum Shape Castings Containing Defects

Fatigue properties of cast aluminum components are controlled by maximum defect size in the material. The larger the maximum defect size, the lower the fatigue strength and life. In the presence of casting defects, crack initiation can be ignored and fatigue life is mainly spent in crack propagation. Therefore, fatigue life of aluminum castings can be predicted by long or short crack growth models. The main problem is defining a starting defect size from readily available data, such as twodimensional (2-D) pore size measurements on metallographic sections. In this article, an extremevalue statistics (EVS) method was used to estimate the maximum defect size in 319 castings from conventional metallographic data. The maximum defect size predicted by EVS agrees quite well with the initiation defect sizes measured from fracture surfaces, and the predicted fatigue life is in reasonable agreement with the experimental data.     

Crystallographic orientation dependence of the growth rate for GaAs low pressure organometallic vapor phase epitaxy

The complete crystallographic orientation dependence of the growth rate for GaAs low pressure organometallic vapor phase epitaxy (LPOMVPE) is determined using a previously described semi-empirical model. A set of LPOMVPE growth rate polar diagrams is presented for reactor temperatures near 550°C as well as near 700°C. Also, the variation of the growth rate polar diagrams as a function of process variables is given. The experimental data utilized in the semiempirical model was attained using a typical horizontal reactor LPOMVPE system and typical LPOMVPE process parameters.