High field electron paramagnetic resonance spectroscopy under ultrahigh vacuum conditions--a multipurpose machine to study paramagnetic species on well defined single crystal surfaces.

A new ultrahigh vacuum (UHV) electron paramagnetic resonance (EPR) spectrometer operating at 94 GHz to investigate paramagnetic centers on single crystal surfaces is described. It is particularly designed to study paramagnetic centers on well-defined model catalysts using epitaxial thin oxide films grown on metal single crystals. The EPR setup is based on a commercial Bruker E600 spectrometer, which is adapted to ultrahigh vacuum conditions using a home made Fabry Perot resonator. The key idea of the resonator is to use the planar metal single crystal required to grow the single crystalline oxide films as one of the mirrors of the resonator. EPR spectroscopy is solely sensitive to paramagnetic species, which are typically minority species in such a system. Hence, additional experimental characterization tools are required to allow for a comprehensive investigation of the surface. The apparatus includes a preparation chamber hosting equipment, which is required to prepare supported model catalysts. In addition, surface characterization tools such as low energy electron diffraction (LEED)/Auger spectroscopy, temperature programmed desorption (TPD), and infrared reflection absorption spectroscopy (IRAS) are available to characterize the surfaces. A second chamber used to perform EPR spectroscopy at 94 GHz has a room temperature scanning tunneling microscope attached to it, which allows for real space structural characterization. The heart of the UHV adaptation of the EPR experiment is the sealing of the Fabry-Perot resonator against atmosphere. To this end it is possible to use a thin sapphire window glued to the backside of the coupling orifice of the Fabry Perot resonator. With the help of a variety of stabilization measures reducing vibrations as well as thermal drift it is possible to accumulate data for a time span, which is for low temperature measurements only limited by the amount of liquid helium. Test measurements show that the system can detect paramagnetic species with a density of approximately 5 × 10(11) spins/cm(2), which is comparable to the limit obtained for the presently available UHV-EPR spectrometer operating at 10 GHz (X-band). Investigation of electron trapped centers in MgO(001) films shows that the increased resolution offered by the experiments at W-band allows to identify new paramagnetic species, that cannot be differentiated with the currently available methodology.

[Results after rebuilding the ossicular chain using the autogenous incus, ionomer-cement-and titanium implants (tympanoplasty type III)]. / Ergebnisse der Tympanoplastik Typ III mit autogenem Amboss sowie Ionomerzement- und Titanimplantaten.

BACKGROUND: A defective ossicular chain can reliably be reconstructed with standardized techniques using e. g. modern alloplastic materials. The comparison of clinical and functional results have proved its worth. Prospective clinical trial as well as collecting and evaluating relevant intraoperative and postoperative findings may be helpful to find the appropriate bone substitute in each case when rebuilding middle ear structures. METHOD: In 354 middle ears (332 patients) the defective or destroyed ossicular chain was rebuilt with the carefully trimmed autogenous incus (n = 83), with ionomer-cement implants (n = 100) and with titanium prostheses (n = 171). The follow-up of the earmicroscopic findings and middle ear function extended over a period of 1.5 years postoperatively on an average (min. 3 months, max. 6 years). The modified otologic record form named "Würzburger Ohrbogen" was used for preoperative and operative data, the "Ohrnachsorgebuch" for the postoperative follow-up. RESULTS: Using incus the air bone gap was improved up to 15 dB in the main speech area. Thus the average remaining conduction deficit was less than 10 dB. The "taking" of ionomer based cement prostheses and titanium prostheses was equally good. The cement implants showed a tendency to protrusion (n = 3), 2 titanium implants were extruded. The air bone gap decreased about 10 to 35 dB using titanium total prosthesis and about 15 to 20 dB using ionomer-cement total prosthesis. The remaining air bone gap with titanium implants was slightly less than with the ionomer-cement PORP (10-15 dB). The air bone gap using the titanium TORP was diminished in a reach of 10 to 35 dB, with the ionomer cement prosthesis between 15 to 20 dB. The remaining gap in the main speech area was slightly favorable to titanium (less than 15 dB) compared with the ionomer-cement TORP. Comparing higher frequencies the air bone gap of titanium was recognizable due to its light weight, but less impressive than expected. Revision surgery (n = 50) has to be performed by reason of cholesteatoma (n = 9), adhesive process (n = 8), dislocation of alloplastic prostheses (n = 8) and because of proposed "second look" (n = 14). CONCLUSIONS: Compared with other materials autogenous implants used for reconstruction of the incus have proved their value, however a deterioration of the sound transmission may develop in the long run. The middle ear compatibility of ionomer-cement implants is similar to titanium implants. The functional results of the titanium implants seem to be slightly superior.

[Detection of nitric oxide synthases in physiological and pathophysiological processes of the nasal mucosa]. / Nachweis von Stickstoffmonoxidsynthasen bei physiologischen und pathophysiologischen Prozessen in der Nasenschleimhaut.

Nitric oxide (NO) can play an important role in the regulation of vascular tone and neurotransmission, as well as in non-specific immunoreactions and inflammation in a variety of tissues. Increased quantities of nitric oxide in respired air can be measured during inflammatory processes. However, the exact role and precise sources of NO under physiological and pathophysiological conditions within the airways remain to be defined. Three isoforms of NO-synthases can be distinguished: two constitutive (neuronal and endothelial) Ca(2+)-dependent cNOS and one inducible Ca(2+)-independent iNOS (NOS II). Constitutive NOS (NOS I and III) release a basal amount of NO under physiological conditions. The inducible form once expressed can catalyse the generation of large quantities of NO. Many kinds of cells, such as macrophages, neutrophils, endothelium and smooth muscle cells, are capable of expressing NOS II. Since all isoforms of NO-synthase seem to be present in nasal tissues and the expression of iNOS under inflammatory conditions seems to be responsible for excessive production of NO, the distribution of NOS-isoforms (especially NOS II) in normal and inflammatory nasal tissue, as well as the exact requirements for expression of iNOS remain to be proven. Non-inflamed fresh human nasal mucosa from the middle turbinate was compared immuno-histologically with nasal mucosa having the typical findings of chronic polypoid rhinosinusitis (i.e., polypoid middle turbinates and polyps of the middle nasal duct). In order to gain more information about the mechanisms of acute inflammation, non-inflamed vital turbinates were incubated in vitro with the proinflammatory substances bacterial lipopolysaccharides (LPS) and tumor necrosis-factor (TNF) for 30, 60, 90, 120, 180 and 240 min. Subsequent to exposure to NADPH-diaphorase and immunostaining with specific antibodies to each NOS-isoform, clearly increased or initiated expressions of inducible NOS (iNOS) in blood vessels, glands, macrophages and epithelium of chronically inflamed and LPS-incubated nasal tissue became apparent in comparison to the non-inflamed controls. In contrast, NOS III/NOS I seemed to be not affected. The onset of immunohistochemically recognizable NOS II expression was observed after 90 min incubation with of LPS/TNF-alpha. Polypoid tissue showed a strong increase in submucosal thickness and a high infiltration of iNOS-positive leukocytes (granulocytes and macrophages) compared to the LPS-incubated non-inflamed specimens. These findings implicate NOS II generated nitric oxide as a key agent for causing swelling, secretion and obstruction in patients with acute and chronic polypoid or allergic rhinitis. These findings also suggest that molecular NO has to be considered in the pathophysiology of chronic polypoid rhinosinusitis.

In vitro expression of inducible nitric oxide synthase in the nasal mucosa of guinea pigs after incubation with lipopolysaccharides or cytokines.

In order to demonstrate the involvement of nitric oxide synthases (NOS)--in particular the inducible isoform (iNOS)--in inflammatory processes within the nasal airways, we used organ-bath incubation to study isolated inferior turbinates and mucosa of the maxillary sinus of guinea pigs. The pattern of the expression in various substructures of the nasal mucosa was of special interest. Mucosa was incubated for 6 h with lipopolysaccharides (LPS) produced by E. coli, interleukin II (IL-2) or tumor necrosis factor-alpha (TNF-alpha). Saline was used as the control solution. Following incubation the specimens were fixed in buffered 4% formaldehyde solution over a period of 4 h. Tissues were next exposed to nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase-reaction and immunostained with specific antibodies to iNOS. Results then showed a clearly increased or initiated expression of iNOS in epithelium, glands, leucocytes and blood vessels of treated tissues in comparison to the control specimens. The inflammatory mediator LPS and the cytokines Il-2 or TNF-alpha alone were found to be capable of increasing the expression of iNOS, although the effects of LPS clearly exceeded those of the cytokines. This finding implicates iNOS-generated nitric oxide as a key factor for causing nasal swelling, secretion and obstruction during nasal infections and allergic episodes.

Serum high molecular weight dipeptidyl peptidase IV (CD26) is similar to a novel antigen DPPT-L released from activated T cells.

This study demonstrates that the 175-kDa form of dipeptidyl peptidase IV (DPPIV) found in normal human serum is identical with a similarly-sized Ag, DPPT-L, found to be rapidly expressed on the surface of activated T cells. As activation progresses, the expression of DPPT-L reaches a peak on day 3, after which expression falls, whereas expression of the 105-kDa CD26/DPPIV detected by the mAb 1F7 increases, as does the ability to bind adenosine deaminase. The loss of DPPT-L from the surface of activated T cells correlates exactly with the appearance of DPPT-L and DPPIV activity in serum-free tissue culture medium. The release of DPPIV was generally greater from CD4+ cells than from CD8+ T cells, and within the CD4+ subset, the CD45RO+ subset was the major source, which correlated with surface expression before culture. We show that the DPPIV released from activated T cells is antigenically, biochemically, and enzymatically similar to DPPIV circulating in the serum and is distinct from the DPPIV activity of 105-kDa CD26. The T cell-released DPPIV is able to function as a costimulating molecule for the response to the recall Ag, tetanus toxoid, at levels similar to those at which recombinant soluble CD26 and serum DPPIV exhibit costimulatory function, suggesting that the released DPPIV may serve an important immunoregulatory function in vivo, both locally and within the systemic circulation.

A novel form of dipeptidylpeptidase IV found in human serum. Isolation, characterization, and comparison with T lymphocyte membrane dipeptidylpeptidase IV (CD26).

Human CD26, a Type II membrane glycoprotein with intrinsic dipeptidylpeptidase IV (DPPIV) activity and ability to bind adenosine deaminase type I (ADA-1), is expressed on epithelial cells constitutively, but on T lymphocytes its expression is regulated. A soluble form of CD26/DPPIV has been described in plasma and related to immunological status, but it has been defined by the presence of DPPIV activity rather than by isolation. Using nondenaturing chromatographic techniques followed by nondenaturing native preparative electrophoresis, we obtained a homogeneous preparation of soluble serum DPPIV and compared it with a recombinant soluble CD26/DPPIV (rsCD26). We show that serum DPPIV is a monomer of 175 kDa in contrast to rsCD26 of 105-110 kDa, that it exists as a trimer, and that it is probably a serine proteinase. Deglycosylation removed N-linked sugar from both serum DPPIV and rsCD26; no O-linked glycosylation was observed, revealing a protein core of 130 kDa for serum DPPIV. The large serum form expresses functional DPPIV activity with substrate and inhibitor specificities and pH activity profile similar to those of rsCD26. Epitope analysis showed that monoclonal antibodies against five epitopes expressed by rsCD26 also bound, but more weakly, with serum DPPIV. Analysis of peptides after limiting proteolysis and N-terminal sequences reveals no homology with rsCD26 but some identity with other peptidases. Unlike rsCD26, the serum form does not bind ADA-1 and has no ADA-1 already associated with it. Similarly to rsCD26, serum DPPIV is a potent T cell costimulator. We conclude that the serum form of DPPIV is unique and is not a breakdown product of membrane CD26. The conservation of DPPIV activity and five epitopes specific to rsCD26 suggest, however, a significant structural similarity.

Promoting occupational therapy by using a simulated hemiplegic arm to demonstrate dressing technique.

When creating a promotional booth to increase awareness about occupational therapy remember that (a) people learn by participating and (b) positive reinforcement encourages a greater sense of learning. In addition, follow these rules: (a) keep the task simple, (b) eliminate jargon and use nonmedical terms, (c) encourage interaction and reward thoughtful reflections, and (d) make sure that participants leave with some promotional materials in hand. As described here participants acquired a hemiplegic arm and tried to dress. They were immediately rewarded with an occupational therapy sticker. Acting as patients, they experienced visual deficits with a special pair of glasses. The occupational therapist encouraged people to ask questions about dressing and visual deficits and about the display of splints and adaptive equipment. People helped themselves to occupational therapy promotional buttons and literature. They left with a basic understanding of hemiplegia. They also understood how an occupational therapist works with a patient to help the patient regain functional independence.

Affective responses to keeping and not keeping an activity product.

This study examined whether keeping or not keeping the products of a craft activity influences affective meaning and mood. Four groups of undergraduate (n = 23) and graduate (n = 20) students created block-printed stationery. Two groups were allowed to keep their stationery, and two groups were not. The measure of mood changes was the Bipolar Profile of Mood States, which subjects completed before and after the activity. After the activity, subjects also completed the Osgood 12-scale short-form semantic differential, which measured the affective meanings of the activity. Data analysis revealed significant differences between conditions on two out of nine variables. The subjects who could not keep their products became significantly more hostile and significantly more energetic than the subjects who kept their stationery. Implications for occupational therapy theory and further research possibilities are discussed.