Pseudomonas keratitis. The role of an uncharacterized exoprotein, protease IV, in corneal virulence

PURPOSE: The role of exoproteins in the pathogenesis of Pseudomonas aeruginosa keratitis was investigated in three animal models by assessing the relationship between corneal virulence and the activities of exotoxin A, elastase, alkaline protease, and an uncharacterized protease, protease IV. METHODS: The four Pseudomonal strains tested included a prototype strain (ATCC 27853) producing exotoxin A, elastase, and alkaline protease; a parent strain (PA103) producing only exotoxin A and protease IV; a mutant (PA103-29) producing only protease IV; and a mutant (PA103-AP1) producing exotoxin A and having only approximately 5% of the protease IV activity of its parent. Corneal virulence was evaluated in the mouse scratch, rabbit scratch, and rabbit intrastromal models in terms of clinical signs (slit lamp examination, slit lamp examination), and viable bacteria. RESULTS: Protease IV, the only protease produced by PA103 and PA103-29, was found to produce a unique band on zymograms (120 kDa) and to react distinctively with a synthetic substrate. Evidence for the role of protease IV in corneal virulence included two findings: PA103-29,which produced protease IV but not the other exoproteins, caused infections that were as severe as those caused by the prototype strain (ATCC 27853) in all three models (P>0.24); and PA103-AP1, the strain deficient in 95% of the parent protease IV activity, mediated infections characterized by slit lamp examination scores significantly lower than those of infections caused by the parent (PA103) or the prototype strain (ATCC 27853) in the rabbit and mouse scratch models (P<0.02). CONCLUSIONS: Protease IV was found to be a novel Pseudomonas protease contributing to corneal virulence in rabbits and mice when infections were initiated at the corneal surface. Furthermore, production of protease IV in low quantities was sufficient for virulence when the topical stages of keratitis were bypassed by an intrastromal injection of Pseudomonas.     

Beanspruchungsverhalten von PVD-CrN Beschichtungen auf Leichtmetallwerkstoffen

Behaviour of light metals with PVD-CrN coatings using different test methods The application of thin hard coatings on machining tools, e.g. drilling tools is state of the art. The increase of lifetime of coated Tools compared to uncoated tools is well known. [1]. In this context ?separation of functions”? is an often used phrase, by meaning the separation of functions of the volume and the surface of materials. Going one step forward, from the point of view of tribology or corrosion this means, you have ?only”? to protect the surface by using a ?good”? coating without looking at the material underneath. In the past the influence of substrate materials or the optimization of the system (substrate-coating) was not the main aim of PVD development. Looking at substrate and coating as a system is especially necessary, if the differences of the properties of substrate and coating are large (e.g. hard coating – light metals). This paper shows different aspect of the tribological and electrochemical characterization of PVD coated light metals (CrN coating).     

Protein thermostability in extremophiles

Thermostability of a protein is a property which cannot be attributed to the presence of a particular amino acid or to a post synthetic modification. Thermostability seems to be a property acquired by a protein through many small structural modifications obtained with the exchange of some amino acids and the modulation of the canonical forces found in all proteins such as electrostatic (hydrogen bonds and ion-pairs) and hydrophobic interactions. Proteins produced by thermo and hyperthermophilic microorganisms, growing between 45 and 110 degrees C are in general more resistant to thermal and chemical denaturation than their mesophilic counterparts. The observed structural resistance may reflect a restriction on the flexibility of these proteins, which, while allowing them to be functionally competent at elevated temperatures, renders them unusually rigid at mesophilic temperatures (10-45 degrees C). The increased rigidity at mesophilic temperatures may find a structural determinant in increased compactness. In thermophilic proteins a number of amino acids are often exchanged. These exchanges with some strategic placement of proline in beta-turns give rise to a stabilization of the protein. Mutagenesis experiments have confirmed this statement. From the comparative analysis of the X-ray structures available for several families of proteins, including at least one thermophilic structure in each case, it appears that thermal stabilization is accompanied by an increase in hydrogen bonds and salt bridges. Thermostability appears also related to a better packing within buried regions. Despite these generalisations, no universal rules can be found in these proteins to achieve thermostability.     

Disturbances of nuclear condensation in human spermatozoa: search for mutations in the genes for protamine 1, protamine 2 and transition protein 1

During spermiogenesis, the successive replacement of the somatic histones by basic proteins, the transition proteins and protamines, allows normal sperm nuclear condensation. It was suggested that disturbances in nuclear condensation may result in male infertility. Here we report the first molecular analysis of the structure of three genes which code for germ cell-specific nuclear proteins, namely protamine 1 (PRM1), protamine 2 (PRM2) and transition protein 1 (TNP1) in infertile men with disturbed sperm chromatin condensation. In 36 infertile men whose spermatozoa showed a positive reaction with aniline blue, which is an indication for the presence of histones in the nuclei, the complete nucleotide sequences of the coding regions and 5' and 3' untranslated regions of the three genes were evaluated. In addition, 10 infertile patients with oligoasthenoteratozoospermia were studied in the same way, as well as nine infertile patients whose spermatozoa showed a reduction of the protamine 2 content. We did not detect any mutation in the three genes in any of the patients. We assume that the disturbances in the sperm chromatin condensation of our patients, and those described in the literature, are not primarily due to mutations in the genes for PRM1, PRM2 and TNP1.     

Quo Vadis, Research Education?

I read with chagrin Donald T. Campbell's (see record 1990-55812-001) comments on my article on the preparation of researchers in psychology. In "Thesis-Antithesis" (American Psychologist, 1966, 21, 781-787; see record 1967-00924-001), I examined some hidden assumptions upon which the thesis requirement rests and attempted to bring into relationship recent ideas in epistemology with earlier recommendations of the Education and Training Board. I suggested that if the future face of psychological investigations is to be more like search and less like research, we shall have to recapture and invest with meaning the personal qualities in the advisory relationship, often lost sight of under the pressure of academic requirements. I claimed that some degree requirements discourage the personal and professional evolution of the future generation of researchers. Campbell's criticism that I recommended the substitution of research assistantships for the thesis requirement leaves me feeling like the guest in a famous restaurant. The waiter wished to know how the guest would like his tournedos bordelaise. "A center cut and tenderloin portion, by all means, quite well done on the outside, then slightly pink, and, finally, seigneuse in the middle, not a very large portion, just right, with some gently braised mushrooms on the side, and only a touch of sauce," said the guest. The waiter nodded with approval, then turned into the kitchen with the usual refrain: "T-bone coming up." What can one do? (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Microwave Rejection Networks

This is an extension of the Bridged-T infinite rejection network idea transformed into microwave concepts. These devices achieve approximately 100db of attenuation with skirt attenuations approximately 5db. The tuning range between 1 KMc and 10 KMc is continuously covered with 4 units at present. Work is continuing to reduce the number of units to 2. Design parameters, performance curves, and application to MILSTD 826 will be discussed.     

Sensory integration of auditory and visual information.

In exp. I with 3 male and 1 female o, the bisensory recognition of simultaneously presented auditory and visual verbal information was measured as a function of auditory and visual recognition. It is shown that bisensory performance is superior to performance predicted by a model that assumes the 2 modalities are processing the information independently. Instead integrative processing is suggested. In previous studies using theory of signal detectability (tsd), independent processing of bisensory presentations of mathematically equivalent stimuli has been shown. Present results suggest that the mathematically equated stimuli in those studies were not cognitively equivalent. Exp. Ii with 3 male os lends support to this notion by (a) using verbal information in a tsd paradigm, and (b) showing that when the stimuli are equivalent the results are consistent with a model in which bisensory processing occurs integratively on a common decision axis. However, when the stimuli to each mode are not equivalent, results are consistent with a model in which bisensory processing occurs independently on separate decision axes. (french summary) (PsycINFO Database Record (c) 2010 APA, all rights reserved)