Humoral indices of stress in rats

Rats, bearing chronic venous cannulas, were subjected to 30 sec of constant current grid shock at 1 of 6 intensities (0, 0.25, 0.5, 1, 2, 4 mA), after being allowed to acclimate to the test chamber overnight. Blood, sampled before and after shock, was assayed for epinephrine, norepinephrine and corticosterone. Peak levels of both catecholamines increased in a stepwise fashion (i.e., monotonically) with increasing magnitude of stress, as reflected by current intensity of foot shock. Plasma corticosterone did not increase monotonically but instead showed similar increases in the 5 groups of rats that actually received shock. These data support earlier work which indicate that plasma corticosterone is not a sensitive index of stress; this is probably the case because of the relatively narrow range of responsiveness of the adrenal cortex to ACTH. In contrast, both plasma catecholamines appear to satisfy some of the requisites for a sensitive visceral index of stress.     

The functional anatomy of the human anterior talofibular ligament in relation to ankle sprains

The anterior talofibular ligament is the most commonly injured ligament in the ankle. Despite considerable interest in the clinical outcome of treatment protocols, we do not know whether the distinctive pattern of localization of the injuries relates to regional differences in the structure and molecular composition of the ligament. To address this issue, ligaments were examined by histology and immunohistochemistry. Differences in the structure of its two attachments (i.e. entheses) were evaluated with quantitative, morphometric techniques, and regional differences in the distribution of collagens, glycosaminoglycans and proteoglycans were determined qualitatively by immunolabelling. Morphometric analyses showed that bone density was less at the fibular attachment, but that enthesis fibrocartilage was more prominent. Immunohistochemistry revealed the presence of a fibrocartilage (containing type II collagen and aggrecan) at the site where the ligament wraps around the lateral talar articular cartilage in a plantarflexed and inverted foot: the fibrocartilage is regarded as an adaptation to resisting compression. We propose that avulsion fractures are less common at the talar end of the ligament because (1) bone density is greater here than at the fibular enthesis, and (2) stress is dissipated away from the talar enthesis by the 'wrap-around' fibrocartilaginous character of the ligament near the talar articular facet.     

Monoclonal antibodies to bovine serum albumin: affinity and specificity determinations

A panel of 12 monoclonal antibodies (MAb) to bovine serum albumin (BSA) was developed and characterized as to their physiochemical and immunological properties. Affinity constants of the MAb varied over a wide range from 10(5) to 10(8) M-1. MAb were assembled into several groups of non- or minimally interacting antibodies by analysis of competitive binding experiments, and BSA domain and subdomain specificities of the MAb were assigned by analysis of results of MAb binding to purified BSA fragments. Further fine specificity delineation was accomplished by examination of cross-reactivity patterns to several mammalian albumins. The data suggest that some of the low affinity MAb recognize sites on different portions of the BSA molecule, indicating that similar epitopes exist on different domains of the BSA molecule.     

Evaluation of the single radial haemolysis (SRH) technique for rubella antibody measurement.

Sera from 1258 individuals have been tested by four laboratories for rubella antibody by both the haemagglutination-inhibition and single radial haemolysis techniques. There was good agreement between the results obtained by the two methods. Although sheep red blood cells were used in the single radial haemolysis plates, no problems were encountered with sera from patients with infectious mononucleosis. The single haemolysis technique was found to be simple, convenient, and reliable, and suited to the rapid screening of large numbers of sera to assess susceptibility to rubella in the context of a vaccination campaign. However, since the technique does not detect anti-rubella IgM, it should not be used as the only test to investigate suspected recent infection.     

The development of fibrocartilage in the rat intervertebral disc

The development of fibrocartilage in rat lumbar intervertebral discs has been correlated with an immunohistochemical analysis of the changing distribution of extracellular matrix components. Disc anlagen were first recognised by embryonic day 14 as segmental cell condensations. By E16, the notochord formed a series of bulges, each representing a future nucleus pulposus, and the annulus fibrosus had differentiated in the disc anlagen. The inner part of the annulus was composed of cartilage which linked that of adjacent vertebral bodies. The outer part was fibroblastic, with layers of parallel fibroblasts. The long axes of the cells in successive layers lay at an angle of approximately 90° to each other. This criss-cross orientation of cells preceded the oriented deposition of collagen fibres to form the lamellae. Disc anlagen were immunolabelled weakly for types I and III collagen, chondroitin 6-sulphate and dermatan sulphate. Later tissue differentiation was marked by the appearance of type II collagen, chondroitin 4-sulphate and keratan sulphate in the inner annulus. These components also appeared in the outer annulus, but only in adult animals, and indicated metaplastic change in the lamellar fibroblasts. Fibrocartilage in the nucleus pulposus was only seen in old animals, and the origin of the tissue was less clear. However, the fibrocartilage cells appeared to be derived from the cartilage end plate and/or from the inner annulus. We conclude that fibrocartilage in the intervertebral disc is derived from several sources and that the radial distribution patterns of extracellular matrix components in the adult disc are explained by the embryonic origins of its parts.     

CD4 monoclonal antibody pairs for immunosuppression and tolerance induction

A pair of rat anti-mouse CD4 monoclonal antibodies (mAb) have been selected which bind to different epitopes of the molecule. Both the mAb are rat IgG2b and show clear synergistic activity in complement lysis in vitro. When injected together in vivo, they exhibit an improved immunosuppressive effect, compared to each antibody alone, on allogeneic graft rejection, humoral responses and on tolerance induction. Limiting dilution analysis indicates that the in vivo depletion of interleukin 2-producing cells is improved using both mAb by 2-3-fold over that obtained with the individual antibodies. As little as 60 ng per mouse of the CD4 antibody pair was sufficient to allow the induction of tolerance to human gamma-globulin, even without elimination of the CD4+ cells. The results suggest that appropriate antibody pairs may be good candidates for effective immunosuppressive serotherapy in man.     

Emerging patterns of cardiac conduction in the chick embryo: waveform analysis with photodiode array-based optical imaging.

Major difficulties investigating the developing cardiac conduction system stem from that the embryonic heart is extremely small (< 2 mm) and cardiac activation is relatively rapid (< 8 msec). The objective of this study was to investigate the electrophysiology of the embryonic chick cardiac conduction system at periseptation stages with a photodiode array-based detection method of optical mapping capable of high spatial and temporal resolution. Previous work indicated that, in chicken embryos, a switch occurs in ventricular activation pattern from immature base-to-apex to mature apex-to-base pattern at the time of ventricular septation. It was our aim to map activation in more detail to identify the active pathway or pathways of atrioventricular conduction at these particular stages. Analysis of preseptated hearts (n = 10) showed that the latest atrial activation took place just above the site of the earliest ventricular activation at the ventral left ventricular base. Analysis of postseptated hearts (n = 11) showed apex-to-base conduction consistent with activation through the maturing His-Purkinje system. Evaluation of hearts during septation revealed a gradual transition of ventricular activation patterns rather than an abrupt "switch." External pacing of preseptated hearts revealed significant slowing of interventricular conduction compared with spontaneous beats (spontaneous, 61.7 cm/sec +/- 9 cm/sec vs. paced, 36.5 cm/sec +/- 10 cm/sec). The more detailed mapping revealed that, before septation, the pattern of activation of the ventricular myocardium is consistent with direct atrial-ventricular myocardial connections at the left lateral atrioventricular junction; however, functional evidence for a preferential conduction pathway within the ventricles was present before septation.     

Introduction of new clones of penicillin-nonsusceptible Streptococcus pneumoniae in Hong Kong

Analysis of penicillin-nonsusceptible Streptococcus pneumoniae (PNSP) isolates in Hong Kong by use of a combination of antibiogram typing, serotyping, multilocus sequence typing, and pulsed-field gel electrophoresis indicated that the dissemination of PNSP was the result of the spread of international clones: variants of the Spain(23F)-1 or Spain(6B)-2 clones were the predominant PNSP isolates from 1994 to 1997 and remained so, but Taiwan(19F)-14 and Taiwan serotype 6B clones were disseminated in Hong Kong in 1999 and 2000. Concomitant changes in antibiotic susceptibility profiles, with the rate of susceptibility to chloramphenicol rising from 10% in the period from 1994 to 1997 to 31% (P < 0.001) in 1999 and 2000, were noted to accompany the shift of clones.