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A simple ELISA is described for quantifying expression of bovine leukaemia virus (BLV) antigens in short-term cultures of peripheral blood lymphocytes (PBL) isolated from infected cattle. The PBL-ELISA demonstrated that antigen expression levels in infected cattle could vary by more than 50-fold. Inoculation of sheep with dilutions of lymphocytes from two BLV-infected cattle, differentiated in the PBL-ELISA by 50 to 100-fold, suggested that antigen expression levels were correlated with infectivity. Haematological data indicated that increased antigen expression in PBL cultures was associated with an increased number of circulating B-lymphocytes, irrespective of whether or not an animal had lymphocytosis. This supported the hypothesis that BLV-infected cattle that are PBL-ELISA positive are more infectious and may present a greater risk of transmitting the disease. The applicability of the PBL-ELISA to a field situation was assessed with 98 BLV-infected cattle from three commercial dairy herds with infection prevalences of 11%, 23% and 47%. Similar percentages (49%, 50% and 52%) of PBL-ELISA positive cattle were identified among those infected cattle available for testing in the three herds. An additional 22 infected cattle from an experimental herd were tested to assess the stability of antigen expression levels over an 8 month period. Fewer (27%) of these cattle were identified as PBL-ELISA positive and antigen expression levels were generally lower than those observed in the commercial herds. Antigen expression levels in the experimental herd remained stable over the period of the study. The potential of the PBL-ELISA to assist in BLV eradication programs by identifying those seropositive cattle with the greatest potential to transmit infection is discussed.

A single-site mutation of the flight-muscle-specific actin gene of Drosophila melanogaster causes a substitution of glutamic acid 93 by lysine in all the actin encoded in the indirect flight muscle (IFM). In these Act88FE93K mutants, myofibrillar bundles of thick and thin filaments are present but lack Z-discs and all sarcomeric repeats. Dense filament bundles, which are probably aberrant Z-discs, are seen in myofibrils of pupal flies, but early in adult life these move to the periphery of the fibrils and are not seen in skinned adult fibres. Consistent with this observation, alpha-actinin and other high molecular weight proteins, possibly associated with Z-discs, are not detected on SDS/polyacrylamide gels or Western blots of skinned adult IFM. The mutation lies at the beginning of a loop in the small domain of actin, near the myosin binding region. However, that the mutant actin binds myosin heads is shown by (1) rigor crossbridges in electron micrographs, (2) the appropriate rise in stiffness when ATP is withdrawn in mechanical experiments, and (3) equal protection against tryptic digestion provided by rigor binding between actin and myosin in both wild-type and mutant fibres. Reversal of rigor chevron angle along some thin filaments reflects reversal of thin-filament polarity due to lattice disorder. The absence of Z-discs, alpha-actinin and two high molecular weight proteins, and binding studies by others, suggest that the substitution at residue 93 affects the binding of the mutant actin to a protein, possibly alpha-actinin, which is necessary for Z-disc assembly or maintenance.

Automated teaching methods permit reliable dissemination of important information to patients. With the professional help of an audiovisual department, a physician can get information to patients in a bright, amusing, and reproducible manner. At the same time, the physician is free to perform other tasks.

Advances in imaging techniques and computer software over the past decade now define brain abnormalities such as tumors in precise, three-dimensional images. We have taken advantage of these technological improvements in designing a system capable of performing magnetic manipulation of an object in a nonlinear trajectory and able to deliver hyperthermia to highly specific targets within the brain. This device relies on external magnets to pull a small metal pellet (thermoceptor) through the brain, and on biplane fluoroscopy to localize the thermoceptor with respect to previously obtained magnetic resonance images. A radiofrequency tuned circuit serves as the hyperthermia applicator and selectively heats the thermoceptor. This paper describes experiments conducted in a series of dogs showing that all three components of the system (magnetic drive, stereotactic real time imaging, and hyperthermia) can be achieved. Integration of the system was accomplished in one animal. These encouraging results need further detailed substantiation in each of the components, yet demonstrate the feasibility of such a device.

A prerequisite for using muscle mutants to study contraction in Drosophila melanogaster is a description of the mechanics of wild-type muscles. Here we describe the mechanics of two different wild-type muscles; the dorsal longitudinal flight muscle which is asynchronous (nerve impulses are not synchronised with each contraction), and a leg muscle, the tergal depressor of the trochanter, which is synchronous. We have compared their mechanics to those of the asynchronous flight and the synchronous leg muscle from the giant waterbug Lethocerus indicus. We found that the mechanics of the asynchronous flight muscles from the two species were similar. At rest length both muscles had a high relaxed stiffness, were partially activated by Ca2+ (low steady-state active tension) and, once activated, had a large delayed increase in tension, which was well maintained, in response to a rapid stretch. The rate constant for the delayed increase in tension was about 10 times greater for D. melanogaster than for L. indicus under the same conditions. The mechanics of the synchronous leg muscles from both species were different from those of the flight muscles and resembled those of other synchronous muscles such as vertebrate striated muscle. At rest length, both muscles had a lower relaxed stiffness than the flight muscles, were fully activated by Ca2+ (high steady-state active tension) and, once activated, had a small delayed increase in tension, which was less well maintained, in response to a rapid stretch. The rate constant for the delayed increase in tension was similar for the leg muscles of both species. The different mechanical properties of the flight and leg muscles must arise from differences in their contractile proteins. The demonstration that satisfactory mechanical responses can be obtained from the small (less than 1 mm long) muscles of D. melanogaster will enable future responses from mutant muscles to be tested.

An ELISA for detecting antibody to the bovine leukaemia virus (BLV) core protein p24 is described. The test uses p24 antigen purified from concentrated cell culture supernate by lectin-affinity chromatography and gel filtration. The sensitivity and specificity of the p24-ELISA for diagnosing BLV infection relative to the gp51 agar gel immunodiffusion test, were 98.1 and 96.7%, respectively. In the event of widespread use of gp51 based vaccines, the p24-ELISA should differentiate effectively between naturally infected and vaccinated animals.

Our laboratories are developing a new technique for delivering localized hyperthermia to deep-seated brain tumors. In this technique, a spherical thermoseed is stereotactically navigated through the brain and tumour tissues via the noncontact application of an external magnetic force. The force required to produce motion of a 3 mm diameter sphere through in vitro brain tissues was measured to be 0.07 +/- 0.03 N. This result was obtained from a series of experiments performed on whole brain specimens extracted from adult canines. Data were also taken with a 3 mm x 3 mm cylinder and a 5 mm sphere. An experimental procedure simulating physiological conditions was developed prior to testing. Evaluations of systematic effects included determinations of the calibration uncertainties, tests of the dependence of the measured force on temperature, and studies of the effects of method of storage of the tissue specimens. The results obtained are compared with (and confirmed by) two different series of experiments performed in vivo on adult canines and with another series of experiments using brain phantom gelatin.

Thirty-four children, age 2-19 years, with brain tumors were treated with surgical resection, irradiation, and a cisplatin (CDDP) containing regimen. Audiologic assessments were conducted prior to each cycle of CDDP to monitor the ototoxic effects of CDDP. Twenty-eight patients with posterior fossa (PF) tumors received 5,040 to 5,650 cGy irradiation to the PF and 0-3,600 cGy to the remainder of the craniospinal (CS) axis. Six patients with supratentorial tumors received 5,140-5,580 cGy to the tumor site and 3,600-4,500 Gy to the remainder of the CS axis. Cycles of CDDP (68 mg/m2), lomustine (75 mg/m2), and vincristine (1.5 mg/m2 weekly for 3 weeks) were given every 6 weeks to 30 children immediately following irradiation, and to 4 at relapse. CDDP was infused over 8 h. Significant hearing loss, defined as a greater than 20-dB change from baseline in the hearing level (HL), occurred in the 250- to 2,000-Hz range in 4 of 29 patients receiving a cumulative dose (CD) of 410 mg/m2, and in 14 of 25 patients receiving a CD of 474 mg/m2. At 4,000 Hz, hearing sensitivity progressed from a HL of 20 +/- 2 dB at a CD of 203 mg/m2 to 31 +/- 6 dB (p less than 0.05) at a CD of 474 mg/m2 (p less than 0.0001).(ABSTRACT TRUNCATED AT 250 WORDS)

Hyperthermia has significant potential as an adjuvant form of brain tumor therapy. Current intracranial hyperthermia methods, however, are limited in their ability to control spatiotemporal thermal distribution. A stereotaxic magnetic movement system that may be capable of heating discrete regions of brain to a preselected temperature is described. With this system, a ferromagnetic object (referred to as a thermoceptor) is directed through the brain by an external drive magnet. Real time thermoceptor position is monitored with biplanar fluoroscopy and superimposed on a preoperative magnetic resonance imaging scan using a computer. Once in position, the thermoceptor can be inductively heated by externally generated radiofrequency signals. Experiments on the magnetic drive and imaging aspects of this system have been conducted in vitro and in vivo. Mechanical studies of cadaver dog brains revealed that a mean force of 0.07 +/- 0.03 N was required to move a 3-mm diameter sphere through brain at a speed of less than 1 cm/15 s. A cranial phantom with mechanical properties similar to brain was constructed of gelatin and Plexiglas. With the use of a "neck loop" design drive magnet with a maximum magnetic field strength of 0.10 T, a 3 x 3 mm cylindrical neodymium iron boron thermoceptor was smoothly directed through the phantom in two dimensions. Additional experiments were conducted with a larger drive magnet in five anesthetized dogs. Neodymium iron boron and samarium cobalt thermoceptors of various shapes and sizes were placed into the cerebral cortex through a burr hole, then directed with the drive magnet. Fluoroscopy was used to follow the thermoceptor movements.(ABSTRACT TRUNCATED AT 250 WORDS)

Changes in the density of eye movement during rapid eye movement (REM) sleep are associated with changes in ventilation and ventilatory response in animals. Recent data in patients with chronic obstructive pulmonary disease suggest that periods of frequent eye movements may be associated with hypoxemia during REM sleep. We have therefore investigated the association between eye movements and ventilation and ventilatory pattern in 10 normal men. Expired ventilation was measured using a pneumotachograph attached to a valved face mask with a dead space of 50 ml and incorporating a peripheral CO2 leak detector. Ventilation was reduced (p less than 0.02) in all stages of sleep compared with that during wakefulness, with no difference between the level of ventilation in each sleep stage (awake, 7.18 +/- 0.43 SEM; Stage 2, 6.47 +/- 0.43; Stage 3/4, 6.45 +/- 0.52; REM sleep, 6.55 +/- 0.47 L/min). During REM sleep, eye movements (EMs) were associated with rapid shallow breathing. Dividing REM into 20-s epochs with or without EMs, EMs were associated with a raised breathing frequency (no EMs, 14.4 +/- 0.4 breaths/min; EMs, 15.8 +/- 0.5 breaths/min; p = 0.01), reduced tidal volume (0.49 +/- 0.03 L; 0.41 +/- 0.03 L; p less than 0.01), and reduced minute ventilation (6.87 +/- 0.45 L; 6.27 +/- 0.51 L; p = 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

The classification, clinical presentation, forms of treatment, and theories of pathogenesis of traumatic myositis ossificans circumscripta are reviewed. A group of 7 patients with this lesion was treated with hematoma aspiration and injection of steroids, lysosomal enzymes, and xylocaine, resulting in significant limitation of disability. The determining factor in the development of this disorder seems to be the response of local soft tissues to trauma rather than the magnitude of the trauma itself which we feel accounts for the success of the form of treatment described.

A protein immunoblot test for detecting antibody to the bovine leukemia virus p24 antigen is described. The test employs a crude antigen preparation derived from concentrated cell culture fluid, and an optimised biotin-streptavidin-peroxidase amplification system for immunodetection. The test is highly specific and is more sensitive than the gp51 agar gel immunodiffusion (AGID) test for detection of BLV antibody in cattle and experimentally infected sheep. In a selected set of 30 field sera from cattle which had given equivocal results in the gp51-AGID test, 21 were positive, 4 were negative, and 5 gave an uncertain result in the p24-immunoblot test.