Assembly of microarrays for genome-wide measurement of DNA copy number.

We have assembled arrays of approximately 2,400 BAC clones for measurement of DNA copy number across the human genome. The arrays provide precise measurement (s.d. of log2 ratios=0.05-0.10) in cell lines and clinical material, so that we can reliably detect and quantify high-level amplifications and single-copy alterations in diploid, polyploid and heterogeneous backgrounds.

Corneal transplantation for inflammatory keratopathies in the horse: visual outcome in 206 cases (1993-2007).

OBJECTIVE: To evaluate the visual outcome of three techniques of corneal transplantation surgery in treating severe inflammatory keratopathies in the horse. DESIGN: Retrospective medical records study. ANIMALS STUDIED: Medical records of 206 horses that received corneal transplantation surgery at the University of Florida Veterinary Medical Center from 1993 to 2007 were reviewed. PROCEDURE: Data collected from the medical records included signalment, types of ocular lesions, type of transplant surgery performed, length of follow-up, complications, and visual outcomes. RESULTS: Full thickness penetrating keratoplasty (PK) was performed in 86 horses for melting ulcers, iris prolapse/descemetoceles, and medically nonresponsive full thickness stromal abscesses (SA). Posterior lamellar keratoplasty (PLK) and deep lamellar endothelial keratoplasty (DLEK) are split thickness penetrating keratoplasties that were utilized for medically nonresponsive deep stromal abscesses (DSA) in 54 and 66 eyes, respectively. The most common postoperative surgical complication was graft rejection and varying degrees of graft opacification. Wound dehiscence and aqueous humor leakage was also a common postoperative problem. A positive visual outcome was achieved for PK, PLK, and DLEK in 77.9%, 98.1%, and 89.4%, respectively. CONCLUSIONS: Corneal transplantation is a tectonically viable surgery in the horse with an overall success rate of 88.5% in maintaining vision when treating vascularized and infected corneal disease in the horse.

The Retinal Research Team at the Eye Foundation Hospital/UAB.

The trustees of the Eye Foundation, Inc. and the Eye Foundation Hospital constitute an independent institution, which is associated with the University of Alabama at Birmingham only in research and resident training. Clarence Blair is the Chairman of the Foundation Board. Harold Skalka, M.D., F.A.C.S., is the Chairman of the Department of Ophthalmology, and his offices are in the Eye Foundation Hospital building. With funds raised largely by Alston Callahan, M.D., F.A.C.S., Director of Development, and with the assistance of fellow trustees, the retina research department has been greatly expanded in the last few years. In this article, the seven prestigious retinal investigators are introduced and short summaries of their research aims are presented. All work toward a better understanding of clinical problems which baffle ophthalmologists, as macular degeneration and diabetic eye disease.

Establishing the collaborative practice committee.

One of the first steps in implementing collaborative practice in the hospital is forming a collaborative practice committee to establish goals, to determine how to attain those goals, and to address any problems that may be encountered. This article offers some suggestions for getting the committee off to a good start.

Mapping motor neurone activity to overt behaviour in the leech: internal pressures produced during locomotion.

Several behaviour patterns have been studied in the leech at both the kinematic and neuronal levels. However, very little is known about how patterns of motor neurone activity map to actual movements. Internal pressure is an essential biomechanical property in this process, being responsible for producing the rigidity and posture that allow the directed delivery of forces produced by muscle contraction. To obtain a better understanding of the biomechanical processes involved in movement of the leech, we have measured the internal pressure of the animal by placing catheters through the body wall and into the gut of intact animals showing normal patterns of behaviour. Each type of behaviour had a characteristic pressure waveform. The elongation phase of crawling produced a rapid increase in pressure that peaked when midbody segments were maximally elongated. The pressure produced during the contraction phase of crawling depended on the type of crawl, only inchworm crawling producing a second peak. Whole-body shortening in response to a head poke also produced a pressure peak, but it had a faster rise time. Swimming produced the largest pressure, which was marked by a large sustained increase that fluctuated phasically with undulations of the body. Dual pressure recordings using two catheters demonstrated that pressure was not uniform along the length of the leech, indicating that the body cavity is functionally compartmentalised. Injecting fluid into the gut via a recording catheter allowed us to determine the effects of increasing internal volume on pressure. In line with previous predictions made using an abstract biomechanical model of the leech hydroskeleton, we found that an increase in the volume caused a reduction in the pressure. We are in the process of constructing a more realistic biomechanical model of the leech, based on actual data reported elsewhere. The results in this paper will provide key tests for refining these models.

Control of antigen presentation by a single protease cleavage site.

Protein antigens require limited proteolytic processing to generate peptides for binding to class II MHC molecules, but the proteases and processing sites involved are largely unknown. Here we analyze the effect of eliminating the three major asparagine endopeptidase (AEP)-processing sites in the microbial antigen tetanus toxin C fragment. The mutant antigen is highly resistant to proteolysis by AEP and crude lysosomal extracts and is dramatically impaired in its ability to be processed and presented to T cells. Remarkably, processing at a single asparagine residue (1219) is obligatory for optimal presentation of many T cell epitopes in this antigen. These studies demonstrate that cleavage at a single processing site can be crucial for effective antigen presentation.

Effects of exercise on cognitive and motor function in chronic fatigue syndrome and depression.

OBJECTIVES: Patients with chronic fatigue syndrome complain of physical and mental fatigue that is worsened by exertion. It was predicted that the cognitive and motor responses to vigorous exercise in patients with chronic fatigue syndrome would differ from those in depressed and healthy controls. METHODS: Ten patients with chronic fatigue syndrome, 10 with depressive illness, and 10 healthy controls completed cognitive and muscle strength testing before and after a treadmill exercise test. Measures of cardiovascular functioning and perceived effort, fatigue, and mood were taken during each stage of testing. RESULTS: Depressed patients performed worst on cognitive tests at baseline. During the treadmill test, patients with chronic fatigue syndrome had higher ratings of perceived effort and fatigue than both control groups, whereas patients with depression reported lower mood. After exertion, patients with chronic fatigue syndrome showed a greater decrease than healthy controls on everyday tests of focused (p=0.02) and sustained (p=0.001) attention, as well as greater deterioration than depressed patients on the focused attention task (p=0.03). No between group differences were found in cardiovascular or symptom measures taken during the cognitive testing. CONCLUSIONS: Patients with chronic fatigue syndrome show a specific sensitivity to the effects of exertion on effortful cognitive functioning. This occurs despite subjective and objective evidence of effort allocation in chronic fatigue syndrome, suggesting that patients have reduced working memory capacity, or a greater demand to monitor cognitive processes, or both. Further insight into the pathophysiology of the core complaints in chronic fatigue syndrome is likely to be realised by studying the effects of exercise on other aspects of everyday functioning.