A protective epitope of Moraxella catarrhalis is encoded by two different genes

The high-molecular-weight UspA protein of Moraxella catarrhalis has been described as being both present on the surface of all M. catarrhalis disease isolates examined to date and a target for a monoclonal antibody (MAb 17C7) which enhanced pulmonary clearance of this organism in a mouse model system (M. E. Helminen et al., J. Infect. Dis. 170:867-872, 1994). A recombinant bacteriophage that formed plaques which bound MAb 17C7 was shown to contain a M. catarrhalis gene, designated uspA1, that encoded a protein with a calculated molecular weight of 88,271. Characterization of an isogenic uspA1 mutant revealed that elimination of expression of UspA1 did not eliminate the reactivity of M. catarrhalis with MAb 17C7. In addition, N-terminal amino acid analysis of internal peptides derived from native UspA protein and Southern blot analysis of M. catarrhalis chromosomal DNA suggested the existence of a second UspA-like protein. A combination of epitope mapping and ligation-based PCR methods identified a second M. catarrhalis gene, designated uspA2, which also encoded the MAb 17C7-reactive epitope. The UspA2 protein had a calculated molecular weight of 62,483. Both the isogenic uspA1 mutant and an isogenic uspA2 mutant possessed the ability to express a very-high-molecular-weight antigen that bound MAb 17C7. Southern blot analysis indicated that disease isolates of M. catarrhalis likely possess both uspA1 and uspA2 genes. Both UspA1 and UspA2 most closely resembled adhesins produced by other bacterial pathogens.     

Radiation hardness of electro-optic asymmetric Fabry-Perot modulators

Analog optical links for LHC experiments are being developed, using electro-optical modulators as front-end transmitters. In the inner trackers, these devices will be subject to a harsh radiation environment, combining ionizing radiation and hadrons, up to 10 Mrads and 10¹⁴ (neutron-equivalent)/cm², over the experiment lifetime. The radiation hardness of the Asymmetric Fabry-Perot Modulator (AFPM) has been tested, with both kinds of radiations, up to LHC total doses and fluences. Results confirm a radiation hardness which qualifies the AFPM for application in the detector front ends.     

PTP-NP, a new member of the receptor protein tyrosine phosphatase family, implicated in development of nervous system and pancreatic endocrine cells

The regulation of protein tyrosine phosphorylation is an important mechanism for developmental control. We describe here a new member of the protein tyrosine phosphatase (PTP) family, called PTP-NP (for neural and pancreatic). The cDNA sequence indicates a receptor-type transmembrane molecule. At early organogenesis, in situ hybridization with a probe for the PTP-NP extracellular region detects expression confined to the region of the developing pancreas, an organ of medical importance, but poorly understood with regard to molecular mechanisms of developmental control. This localized expression appears early, even before morphological differentiation of the pancreas, and is found in presumptive precursors of the endocrine cells by the earliest times that they can be distinguished. In neural development, an alternate RNA with a different or missing extracellular region is expressed transiently at early stages of neurogenesis and the full-length PTP-NP RNA appears later. To search for a ligand of PTP-NP, a fusion protein probe was made with the extracellular domain fused to an alkaline phosphatase tag. This probe bound strongly to pancreatic islets, providing evidence for a ligand-receptor interaction that could be involved in endocrine cell regulation. The results show PTP-NP is an especially early marker for pancreatic development and suggest it may be a receptor that could control the development of pancreatic endocrine cells.     

Interleukin-1 beta and tumor necrosis factor-alpha stimulate the cat-2 gene of the L-arginine transporter in cultured vascular smooth muscle cells

The production of nitric oxide (NO) from L-arginine by nitric oxide synthase (NOS) in cytokine-stimulated vascular smooth muscle cells (VSMC) is thought to play an important role in the pathophysiology of several vascular disease states including septic shock. This study examines the relationship between cytokine-stimulated NO production and L-arginine transport in cultured VSMC. Cultured VSMC from rat aorta were stimulated with interleukin-1 beta, tumor necrosis factor-alpha, and/or angiotensin II (Ang II); and the accumulation of nitrite, a stable product of NO metabolism, in the culture media and the rates of net L-arginine uptake were measured. Interleukin-1 beta and tumor necrosis factor-alpha, alone or in combination, stimulated both the uptake of L-arginine and the accumulation of nitrite in the culture media in a dose-dependent manner. Inhibition of NOS activity by substituted analogues of L-arginine had no effect on cytokine-stimulated L-arginine transport. Ang II in the presence of cytokines up-regulated L-arginine transport while inhibiting nitrite accumulation. Two forms of the L-arginine transporter, cat-1b and cat-2, are expressed in VSMC. Northern analysis revealed that the cytokine-stimulated increase in L-arginine transport coincided with increased levels of cat-2 mRNA. In contrast, cat-1b does not appear to be regulated by cytokines at the mRNA level, although significant increases in response to Ang II were observed. These results show that, while cytokines can stimulate both NOS activity and L-arginine uptake, NO production is not required to signal the increase in L-arginine transport. Furthermore, Ang II and cytokine stimulation of L-arginine uptake involves the differential regulation of the cationic amino acid transporter (cat) genes.     

ICAM-1 induction in the mouse CNS following irradiation

Injury to the central nervous system (CNS) results in inflammation, increased trafficking of leukocytes into the CNS, induction of cytokines, and exacerbation of the primary injury. The increased trafficking of neutrophils into the CNS has been described following a number of injury models including stab, stroke, and excitotoxin-induced injury. This enhanced trafficking has largely been ascribed to the adhesion molecule intercellular adhesion molecule-1 (ICAM-1, CD54). In the current study, we wished to determine if the inflammation caused by irradiation of the CNS resulted in a similar induction of ICAM-1. C3H/HeJ mice were irradiated using gamma irradiation aimed over the right cerebral hemisphere. The relative induction of ICAM-1 mRNA levels was determined using quantitative RT-PCR 6 hours following irradiation with either 0, 5, 15, 25 or 35 Gy. ICAM-1 message was seen to exhibit a normal dose response curve with increasing mRNA levels seen at 15 Gy and higher. To determine the cellular distribution of the ICAM-1 protein following irradiation, mice were sacrificed at 4 hrs, 24 hrs, 48 hrs and 7 days following 25 Gy irradiation and the tissue was processed for ICAM-1 immunocytochemistry. ICAM-1 staining was seen to increase in both endothelial cells and astrocytes beginning as early as 4 hrs. The staining intensity continued to increase throughout the 7 day period observed. Together, these results suggest that irradiation of the CNS causes a rapid induction of both ICAM-1 mRNA and protein. This suggests that increased leukocyte trafficking into the CNS may exacerbate the inflammation induced by radiation injury.     

Contrast medium enhanced magnetic resonance angiography. Theory, technique and practical implementation

Contrast enhanced (CE) magnetic resonance angiography affords angiographic depiction of extended vascular territories with high quality and diagnostic value. A prerequisite is the fast acquisition of a three-dimensional gradient-echo data set during the injection of a bolus of a T1-shortening contrast agent. We describe the dependence of the quality of CE-MRA on technical parameters of different MR-scanners and consider some fundamental facts and practical guidelines concerning the contrast agent injection.     

Interspecies differences and variability with time of protein precipitation activity of extractable tannins,crude protein,ash, and dry matter content of leaves from 13 species of Nepalese fodder trees

Dry matter, ash, crude protein, and protein precipitation activity (PPA) of 13 Nepalese tree fodder species were monitored in dried samples prepared monthly between November 1990 and May 1991, and additionally in November 1991, covering the season when they are particularly important as fodder. Monthly levels of dry matter, ash, and crude protein were fairly stable except when there was new leaf growth, although year to year differences in dry matter were found inBrassaiopsis hainla (Bh),Dendrocalamus strictus (Ds),Ficus roxburghii (Fr), andQuercus semecarpifolia (Qs). Tannin PPA fluctuated considerably inArtocarpus lakoocha (Al),Ficus glaberrima (Fg),F. nerrifolia (Fn), Fr,F. semicordata (Fs),Litsea polyantha (Lp), andPrunus cerasoides (Pc), and to a lesser extent in Bh,Castanopsis indica (Ci),C. tribuloides (Ct),Quercus lamellosa (Ql), and Qs. Similar fluctuations in PPA were observed in fresh leaf samples taken weekly. Ds did not have any detectable PPA. Trends in PPA fluctuation were generally similar for trees located at similar altitudes. Fr, Pc, Al, Fn, Ql, and Ci had falling PPAs before shedding leaves. Some of the fluctuations in Fr, Fs, Fg, Pc, and Lp were apparently due to changes in the extractability and quantity of condensed tannins. These fluctuations in PPA may affect the nutritive value of the fodders.     

Purification and crystallisation of the autoantigen thyroid peroxidase from human Graves' thyroid tissue

Milligram quantities of the human membrane autoantigen thyroid peroxidase (TPO) have been purified to a high degree of homogeneity by a combination of detergent solubilisation, monoclonal antibody affinity, and ion exchange chromatography, from pooled Graves' disease thyroid glands. The purified TPO of greater than 90% purity was enzymatically active as judged by its ability to oxidise guaiacol. Crystals of TPO have been grown from solutions of the protein solubilised in sodium deoxycholate, in the presence of ammonium sulphate. The crystals exhibited birefringence under polarised light, indicative of molecular order. Crystallisation of this large, membrane autoantigen represents the first step in delineating the complete three-dimensional structure of a human autoantigen involved in destructive thyroiditis.