Cytokine-directed therapies for the treatment of chronic airway diseases

Multiple cytokines play a critical role in orchestrating and perpetuating inflammation in asthma and chronic obstructive pulmonary disease (COPD) and several specific cytokine and chemokine inhibitors now in development as future therapy for these diseases. Anti-IL-5 antibody markedly reduces peripheral blood and airway eosinophils, but does not appear to be effective in symptomatic asthma. Inhibition of IL-4 despite promising early results in asthma has been discontinued and blocking IL-13 might be more effective. Inhibitory cytokines, such as IL-10, interferons and IL-12 are less promising, as systemic delivery produces side effects. Inhibition of TNF-alpha may be useful in severe asthma and for treating severe COPD with systemic features. Many chemokines are involved in the inflammatory response of asthma and COPD and several small molecule inhibitors of chemokine receptors (CCR) are in development. CCR3 antagonists (which block eosinophil chemotaxis) and CXCR2 antagonists (which block neutrophil and monocyte chemotaxis) are in clinical development for asthma and COPD, respectively. Because so many cytokines are involved in asthma, drugs that inhibit the synthesis of multiple cytokines may prove to be more useful; several such classes of drug are now in clinical development and any risk of side effects with these non-specific inhibitors may be reduced by the inhaled route.     

Ionic channels of the inner segment of tiger salamander cone photoreceptors

Cone photoreceptors were isolated enzymatically and their ionic currents studied by the whole-cell, gigaseal voltage-clamp technique. Five nonsynaptic currents were identified. A prominent, poorly selective cation current, Ih, activated after a delay during hyperpolarizations and then deactivated with a delay on return to potentials greater than -50 mV. An empirical model for Ih gating kinetics is developed with three open and two closed states. Depolarization elicits a small, voltage-gated calcium current (ICa). Block by nitrendipine, nickel, cadmium, and cobalt, increase of current with barium, lack of rapid inactivation, and relatively high threshold suggest an L-type Ca channel. No evidence was found for low-threshold Ca channels. An anion current ICl(Ca) was present after pulses that led to a significant inward ICa (but not IBa) and was not elicited when cobalt was present. Tails of ICl(Ca) were short (100 ms) after short depolarizations and were longer after longer depolarizations. Two TEA-sensitive K currents were also elicited by depolarizations. One, IK(Ca), was calcium sensitive. We looked for modulation of Ih, ICa, and ICl(Ca) by a number of neurotransmitters. No changes of Ih were seen, but ICa and ICl(Ca) were depressed in a few cones when GABA or adenosine were applied. We discuss how this modulation might contribute to the feedback effects of horizontal cells on cones when surrounding cones are illuminated.     

Simultaneous freeze tolerance and avoidance in individual fungus gnats, <Emphasis Type="Italic">Exechia nugatoria</Emphasis>

Freeze tolerance and freeze avoidance are typically described as mutually exclusive strategies for overwintering in animals. Here we show an insect species that combines both strategies. Individual fungus gnats, collected in Fairbanks, Alaska, display two freezing events when experimentally cooled and different rates of survival after each event (mean ± SEM: −31.5 ± 0.2°C, 70% survival and −50.7 ± 0.4°C, 0% survival). To determine which body compartments froze at each event, we dissected the abdomen from the head/thorax and cooled each part separately. There was a significant difference between temperature levels of abdominal freezing (−30.1 ± 1.1°C) and head/thorax freezing (−48.7 ± 1.3°C). We suggest that freezing is initially restricted to one body compartment by regional dehydration in the head/thorax that prevents inoculative freezing between the freeze-tolerant abdomen (71.0 ± 0.8% water) and the supercooled, freeze-sensitive head/thorax (46.6 ± 0.8% water).     

The Occurence and Properties of Uric Acid Decomposing Anaerobic Bacteria in the Avian Caecum

S ummary . Anaerobic bacteria which decompose uric acid have been found in the caeca of chickens, turkeys, ducks, pheasants and guinea-fowl at levels between 5.4 × 10⁸ and 1.8 × 10¹⁰/g of caecal material (wet weight). A study has been made of the properties and identity of 35 strains which were present in high numbers in the chicken caecum. The isolates included strains of Bacteroides clostridiiformis var. clostridiiformis, Fusobacterium plauti, Clostridium malenominatum, Peptostreptococcus productus and a number of types which could not be identified with known species. Of these 3 were Bacteroides spp., 3 Eubacterium spp., 2 Peptostreptococcus spp. and 1 was an anaerobic Streptococcus . None of the strains had an absolute requirement for uric acid.     

Study of Dyspepsia in a General Practice as Assessed by Endoscopy and Radiology

Fifty consecutive patients presenting in a general practice with dyspepsia were studied. Each patient was referred for a combined barium-meal examination and cholecystogram, followed by fibreoptic endoscopy of the oesophagus, stomach, and duodenum. Thirty patients had a specific lesion of the upper digestive tract and a further four had gall-bladder disease; of these, 16 required surgical treatment. Endoscopy in the remaining 16 patients showed nine with mucosal abnormalities, leaving only seven patients to be classified as normal.The advantage of having an endoscopy service to supplement radiological investigation of dyspepsia is shown.     

The distribution of satellite and main-band DNA components in the melanogaster species subgroup of Drosophila

Fractionation of total adult DNA of five of the seven species of the melanogaster species sub-group of Drosophila in actinomycin D and distamycin A caesium density gradients has revealed the presence of three main-band DNA components, common to all species, and ten satellite DNAs that are distributed between the species. Satellite DNAs are either unique to a species or common to two or more species. The abundance of a common satellite DNA varies between species. There is no simple relationship between the presence of a satellite DNA and a branch point of phylogenetic divergence; nevertheless the arrangement of the species in a phylogeny that is based on the numbers of satellites held in common accurately reflects the pattern of relationships between the same species based on differences in inversions of polytene chromosomes. The species can be similarly arranged according to the compositions of their mitochondrial DNAs. It is possible that the same basic set of sequences, each of low frequency, is common to all species with arbitrary or selected amplification of particular sequences to differing extents in individual species. The conservation of satellites in the group and the close parallel between the distributions of satellites and inversions between the species suggests that either the processes that operate to change both chromosomal phenomena are similarly time-dependent and occurring at relatively low rates or that their rates of change are restricted according to some undetermined functions of these aspects of the genome.     

Detecting estrus in synchronized heifers-using tailpaint and an aerosol raddle

A synchronization treatment was initiated when each of 1227 heifers (four trials) was tailpainted. The tailpaint was sprayed with an aerosol raddle at the end of the treatment period. The heifers were in herds of 20 to 279 animals. Each herd was observed for estrus at selected post treatment intervals. A heifer was considered to be (or to have been) in estrus when the raddle was rubbed off. In three of the trials, animals which had the raddle removed were inseminated at 48h following the end of the synchronization treatment. The tailpaint of an inseminated animal was scored from 0 (less than 10% of the paint remained) to 5 (more than 90% of the paint remained) and was then reraddled with a second color. The detection-insemination sequence was always repeated at 72 and 96h, and sometimes at 120h. Animals which had been previously inseminated, but then had paint scores reduced by at least 2 units were reinseminated 24h later. Over the four trials, 94.5% of the heifers were detected in estrus through the use of the tailpaint and raddle system. The remaining 67 animals included only 10 (0.8%) which had ovulated without being detected in estrus. The reinsemination rate on consecutive days was 11.3% and was highest among animals that had a tailpaint score of 4 or 5 at 48h. The proportion of animals detected in estrus at selected posttreatment intervals varied with the different synchronization treatments used within one herd, or with the same treatment used in different herds. The combination of tailpaint, raddling, tailpaint scoring and reraddling is a simple sequence which can be effectively used to detect estrus among heifers synchronized in research or commercial herds.     

Autoradiographic localization of calcitonin gene-related peptide (CGRP) binding sites in human and guinea pig lung

125I-Human calcitonin gene-related peptide (hCGRP) binding sites were localized in human and guinea pig lungs by an autoradiographic method. Scatchard analysis of saturation experiments from slide-mounted sections of guinea pig lung displayed specific 125I-hCGRP binding sites with a dissociation constant (Kd) of 0.72 +/- 0.05 nM (mean +/- S.E.M., n = 3) and a maximal number of binding sites (Bmax) of 133.4 +/- 5.6 fmol/mg protein. In both human and guinea pig lung, autoradiography revealed that CGRP binding sites were widely distributed, with particularly dense labeling over bronchial and pulmonary blood vessels of all sizes and alveolar walls. Airway smooth muscle and epithelium of large airways was sparsely labeled but no labeling was found over submucosal glands. This localization corresponds well to the reported pattern of CGRP-like immunoreactive innervation. The findings of localization of CGRP binding sites on bronchial and pulmonary blood vessels indicate that CGRP may be important in the regulation of airway and pulmonary blood flow.     

Biomechanical basis of optimal scoliosis surgical correction

For an optimal approach to surgical correction of scoliosis, it was deemed desirable to biomechanically simulate the set of corrective forces applied by alternative internal fixation systems, so as to determine and apply the internal fixation system producing the best correction under safe levels of forces applied by the fixation systems to the spinal structures. To this end, we have developed, and presented here, (1) a spinal finite-element model relating the applied corrective forces to the corrected spinal configurations, (2) a method for determining the stiffness of the patient's spine prior to surgery, (3) computerized finite-element analysis simulation of alternative internal correction-fixation systems, so as to determine the most efficacious system, (4) instrumentations for surgically implementing the recommendations of the surgical simulation analysis and (5) comparisons of the model-simulated and surgically-obtained corrected spinal configurations. These procedures together constitute the biomechanical foundations of scoliosis surgical correction.     

Failure of highly purified lysyl hydroxylase to hydroxylate lysyl residues in the non-helical regions of collagen.

The activity of highly purified lysyl hydroxylase towards lysyl residues within both the helical and the N-terminal non-helical telopeptide regions of chick type I collagen has been examined. The peptides alpha 1(I)-CB1 and alpha 2(I)-CB1, isolated from protocollagen following CNBr digestion and containing the N-terminal telopeptidyl lysyl residues, failed themselves to act as substrates. With protocollagen as substrate, analysis of products obtained following bacterial collagenase digestion of the reaction mixture showed that overall 37% hydroxylation of lysyl residues within the helical region of collagen had been obtained, which may be maximal. No hydroxylation, however, of the single lysyl residue in either alpha 1(I)-CB1 or alpha 2(I)-CB1, isolated following CNBr digestion of the reaction mixture, was observed, despite the known susceptibility of these residues to hydroxylation. These findings provide strong circumstantial evidence for the suggestion that a lysyl hydroxylase specific for the telopeptidyl residues and distinct from that active towards lysyl residues in the helical portion of the molecule may exist [Barnes, Constable, Morton & Royce (1974) Biochem. J. 139, 461-468].