The use of the polymerase chain reaction to map CD4+ T cell epitopes.

CD4+ T cells recognize processed exogenous antigen in the form of peptides bound to syngeneic major histocompatibility complex class II molecules on antigen-presenting cells. We have developed a novel and convenient method to synthesize and map CD4+ T cell epitopes of cloned antigens using polymerase chain reaction (PCR)-directed construction of genes expressing recombinant protein fragments. Unique restriction sites incorporated into the PCR primers were employed for the unidirectional cloning of gene fragments into a bacterial expression vector that can be induced to high-level expression. The bacterial lysate could be used directly in T cell proliferation assays. Overlapping recombinant fragments spanning the entire protein were generated and tested. The length of the sequence containing the epitope was further reduced by utilizing PCR to generate 3' truncations. Finally, a small number of overlapping peptides spanning a sequence of 39 amino acids were synthesized to identify a thirteen-amino acid peptide epitope within chicken transferrin that stimulates the T helper cell clone D10.G4.1. PCR-directed construction of fragments of antigen allows for optimal design of strategies for the mapping and analysis of CD4+ T cell epitopes.     

Inverting a Laplacian topography map

Summary There are some occasions when it is better to use the Laplacian of the scalp voltages (e.g., for topographic displays and dipole source localization) and other occasions when it is better to use the original voltages (e.g., for comparison to non-Laplacian topography maps). This paper presents a simple algorithm for going between these two representations of the data. The inverse Laplacian involves iterative feedback. An Excel spreadsheet implementation of the algorithm is presented.This research was supported by grant AFOSR 89-0238 from the Air Force Office of Sponsored Research.     

Potential interference of agents on radioiodide thyroid uptake in the euthyroid rat.

The objective of this research was to investigate the merits of controlled studies with euthyroid rats as a means of determining the influence of dose and time after administration of agents that may interfere with radioiodide uptake in the thyroid. METHODS: Potassium iodide (KI), propylthiouracil (PTU), diatrizoate meglumine, and iohexol were selected to represent interfering agents. Two dose levels per agent were investigated. Doses used were 1 and 2 mg/kg of body weight for KI, 3.5 and 7 mg/kg of body weight for PTU, 1 mL/kg (282 mg I/kg) and 2 mL/kg (564 mg I/kg) of body weight for diatrizoate meglumine, and 1 mL/kg (300 mg I/kg) and 2 mL/kg (600 mg I/kg) of body weight for iohexol. The 24-h radioiodide thyroid uptake was determined after (131)I was given at 1, 8, 15, and 22 d after administration of interfering agents. RESULTS: The percentage radioiodide uptake value for the thyroid decreased significantly compared with controls for all agents and both doses on day 1 but returned to control levels by day 22 for all agents and both doses The time to return to normal varied between agents and doses. CONCLUSION: We conclude that the interfering agent, the dose given, and the length of time after administration influence the potential for an agent to affect radioiodide uptake in the thyroid. Further studies with the rat, preferably hyperthyroid, would be beneficial in generating data to reduce confusing contradictory information on the length and severity of interference of agents in radioiodide thyroid studies.     

Functional and morphologic endothelial damage in rabbit external jugular veins stored in heparinized normal saline.

Previous studies have demonstrated that vein storage in normal saline leads to significant mechanical morphological, and biochemical aberrations. However, little information is available regarding the functional damage that occurs. The purpose of this study was to evaluate the effect of saline storage on venous smooth muscle and endothelial function. Segments of ten external jugular veins from male New Zealand White rabbits were placed nondistended in either modified Krebs solution at 37 degrees C (Krebs-stored, KS) or heparinized normal saline at room temperature (saline-stored, SS) for 1 h. Segments 4 mm in length were then simultaneously studied in vitro under isometric tension. There was no difference in maximum tension or sensitivity to either bradykinin or histamine. Acetylcholine-induced relaxation in KS segments was not significantly different from relaxation in a historical cohort of nonstored segments (nonstored 87.4 +/- 1.0% vs. KS 84.5 +/- 2.0%; p = NS). However, there were significant attenuations in SS segment endothelium-dependent relaxation in response to both acetylcholine (KS 84.5 +/- 2.0% vs. SS 76.4 +/- 2.7%, p less than 0.02) and adenosine diphosphate (KS 47.9 +/- 2.9% vs. SS 40.6 +/- 3.7%, p less than 0.002). Relaxant responses to sodium nitroprusside (endothelium-independent) were not significantly different in the two groups (KS 94.6 +/- 1.6% vs. SS 95.7 +/- 2.2%; p = NS). Electron microscopic evaluation of SS segments revealed endothelial cell disruption with cellular edema and loss of intact junctions.(ABSTRACT TRUNCATED AT 250 WORDS)     

THE POINT PREVALENCE OF WOUNDS IN A TEACHING HOSPITAL

Background : Wounds are a common problem, particularly in the elderly population. The scale of wound problems in hospital is largely unknown because wounds are widely dispersed. The present study examined the point prevalence of hospital wounds and undertook a pressure ulcer risk assessment of all patients on one day. Methods : All 360 inpatients were surveyed and thoroughly examined. A risk scale for pressure sore development, the Norton score, was applied. When wounds were found, information was collected to determine their aetiology. Results : Forty leg ulcers, 40 pressure sores, 85 surgical wounds and seven other types of wounds were found. Most leg ulcer and pressure sore cases were admitted for other reasons. The Norton score did not predict all cases of pressure ulceration. A total of 52% of wounds did not qualify for additional funding under current funding criteria. Conclusion : The prevalence of non-surgical wounds in Heidelberg Repatriation Hospital was easily underestimated. Wound care management can be optimized by staff education and protocol design, early identification of troublesome wounds and of at-risk patients, and a cross-sectional approach that incorporates wound-management teams.