Optical activity and conformation of cobra neurotoxin

Cobra neurotoxin from Formosan cobra (Naja naja atra) venom is a compact globular protein having an intrinsic viscosity of 4.5 mL/g. The protein is stable in 7.5 M urea but can be denatured in 4.1 M guanidine hydrochloride or at elevated temperature (above 70 degrees C). Its conformation remains virtually the same in solvents of lower polarity than water such as 1,2-ethanediol or a mixed solvent of 1-propanol-1,2-ethanediol-water (5:1:1 by volume). The circular dichroism spectrum is "atypical" in water in that the peptide chromophores show a small negative circular dichroic (CD) band at 215 nm, a large positive one at 199 nm, and another large negative one below 190 nm. The CD pattern resembles to some extent that of a beta form but differs in both positions and magnitudes from the latter. It agrees qualitatively with the theoretical calculations of the reverse beta bends, suggesting that cobra toxin contains a considerable amount of beta turns and possibly a mixture of beta form and beta turns.     

Activation of particulate guanylyl cyclase by Vibrio vulnificus hemolysin

Recently we reported that Vibrio vulnificus hemolysin, an exotoxin produced by V. vulnificus, dilates rat thoracic aorta via elevated cGMP levels without affecting nitric oxide synthase. We investigated the mechanism further by observing the guanylyl cyclase activities in cytosolic, membrane, unfractionated, or reconstituted preparations. Hemolysin did not activate guanylyl cyclase in the membrane or cytosolic fraction, while it activated guanylyl cyclase in unfractionated or reconstituted preparation. The increased activity was not inhibited by the HS-142-1, a microbial polysaccharide which antagonizes atrial natriuretic peptide receptor, or 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), a soluble guanylyl cyclase inhibitor. However, it was attenuated by 6-(phenylamino)-5,8-quinolinedione (LY 83.583), which inhibits the catalytic domain of both guanylyl cyclases, and by cholesterol, which blocks hemolysin-incorporation into the membrane. Removing ATP, a cofactor of particulate guanylyl cyclase, attenuated the activation and ATPgammaS, a non-phosphorylating analog, restored it. These results suggest that V. vulnificus hemolysin activates particulate guanylyl cyclase via hemolysin incorporation into the vascular smooth muscle cell membrane in cooperation with certain unidentified cytosolic component(s).     

Catechol O-methyltransferase inhibitor tolcapone has minor influence on performance in experimental memory models in rats

DNA-based immunization is a promising new technique for generating antibodies in laboratory animals for diagnostic purposes in biological science. The main advantages are the elimination of time and labor and the technically demanding steps of antigen purification. The DNA sequence of the protein of interest, cloned in a suitable in vivo expression vector that is administered intramuscularly or intradermally, is sufficient to induce an immune response in animals. We report the induction of antibodies to tobacco mosaic virus (TMV) coat protein (CP) as a highly immunogenic structural protein and potato virus Y (PVY) P1 protein (P1) as a nonstructural protein. The appropriate nucleotide sequences were introduced in a mammalian expression vector (pSG5) and injected intramuscularly into New Zealand White rabbits (Oryctolagus cuniculus). By 10 days post-injection (dpi) a specific immune response was detected against TMV-CP, while it took about 5 weeks for a response to PVY P1. In both cases the antibody titers were significantly above the corresponding pre-immune serum, however, they were considerably below the titer of the matching conventionally produced antiserum. To our knowledge, this is the first report of DNA-based immunization in order to generate antibodies to plant viral proteins, but further improvements are necessary to increase antibody titers before this promising new technique can be introduced broadly in plant science for diagnostic purposes.     

Randomized controlled trial of open and closed haemorrhoidectomy

BACKGROUND: This study was a prospective randomized comparison of healing following open and closed haemorrhoidectomy. METHODS: Sixty-seven consecutive patients (mean(s.e.m.) age 45(1.7) years; 35 men, 32 women) with three prolapsed piles were randomized to open haemorrhoidectomy (n = 34) or closed haemorrhoidectomy (n = 33). RESULTS: Mean(s.e.m.) follow-up was 8.7(0.2) months. There were no differences in the linear analogue pain scores, analgesic requirements and length of hospitalization after open haemorrhoidectomy and closed haemorrhoidectomy. Complete wound healing took significantly longer after closed haemorrhoidectomy (mean(s.e.m.) 6.9(0.7) weeks) compared with open haemorrhoidectomy (4.9(0.4)weeks) (P < 0.05). This was related to wound dehiscence in eight patients. Complication rates, however, were similar except for prolonged serous discharge from unhealed wounds. The anal manometry findings after both procedures were equivalent. CONCLUSION: Open haemorrhoidectomy leads to faster and more reliable wound healing, although this did not result in less pain or fewer complications.     

Nitric oxide-synthesising neurons in the central subnucleus of the nucleus tractus solitarius provide a major innervation of the rostral nucleus ambiguus in the rabbit

We describe an intramedullary nitric oxide synthase (NOS) neural pathway that projects from the nucleus tractus solitarius (NTS) to the rostral nucleus ambiguus (NA) in the rabbit. With the use of NADPH diaphorase histochemistry and NOS immunohistochemistry, a compact group of NOS-positive perikarya was identified in the central subnucleus of the NTS dorsomedial to the tractus solitarius and rostral to the obex. A dense network of NOS terminals was seen in the rostral NA. We investigated whether NOS terminals in the NA derive from NOS perikarya in the central NTS and whether the central NOS pathway links esophageal afferents and efferents. In some rabbits, the central NTS was unilaterally lesioned. In others, Phaseolus vulgaris-leucoagglutinin (PHA-L) was injected into the central NTS, or cholera toxin-gold was injected into the NA, or cholera toxin-horseradish peroxidase (HRP) was injected into the wall of the esophagus. The medulla was subsequently processed to demonstrate PHA-L, cholera toxin-gold, HRP, and NOS reactivity. Seven days after the NTS lesion, we observed a marked decrease in the density of NOS terminals in the ipsilateral NA. After injection of PHA-L into the central NTS, a dense group of PHA-L fibres was seen in the rostral NA, principally ipsilaterally. Afferent fibres from the esophagus were found around the NOS cell bodies in the central NTS, and many of these NOS neurons were double labeled with cholera toxin-gold after injection of this tracer into the NA. NOS terminals were found around NA neurons that were retrogradely labelled from the esophagus. We conclude that the NOS neurons in the central NTS act as interneurons in a central pathway connecting esophageal afferents and efferents.     

Gastrointestinal involvement in Henoch-Sch?nlein syndrome: CT findings

OBJECTIVE: The purpose of this study was to describe the CT features of gastrointestinal involvement in seven patients with Henoch-Sch?nlein syndrome. CONCLUSION: Although the incidence of Henoch-Sch?nlein syndrome is low, it should be considered when CT scans show multifocal areas of bowel-wall thickening, mesenteric edema, vascular engorgement, and nonspecific lymphadenopathy. It should be considered especially in young patients with acute gastrointestinal symptoms.     

Computerized detection of masses in digitized mammograms using single-image segmentation and a multilayer topographic feature analysis

RATIONALE AND OBJECTIVES: We developed and evaluated a computer-aided detection (CAD) scheme for masses in digitized mammograms. METHODS: A multistep CAD scheme was developed and tested. The method uses a technique of single-image segmentation with Gaussian bandpass filtering to yield a high sensitivity for mass detection. A rule-based multilayer topographic feature analysis method is then used to classify suspected regions. A set of 260 cases, including 162 verified masses, was divided into two subsets; one set was used to set the rule-based classification and one was used to test the performance of the scheme. RESULTS: In a preliminary clinical study, the implemented detection scheme yielded 98% sensitivity with a false-positive detection rate of less than one false-positive region per image. CONCLUSION: Single-image segmentation methods seem to have high sensitivity in selecting true-positive mass regions in the first stage of a CAD scheme. A multilayer topographic image feature analysis method in the second stage of a CAD scheme has the potential to significantly reduce the false-positive detection rate.