A method for isolation of sequences missing in one of two related genomes

We describe a novel technique for isolation of sequences that are present in one genome (tracer), but absent in another (driver). Tracer DNA, cleaved with Sau 3A and capped with a single stranded PCR adapter, is allowed to hybridize with an excess of sheared biotinylated driver; biotinylated DNA and its hybrids with the tracer are removed by phenol/chloroform extraction after incubation with streptavidin. After several rounds of subtraction the ends of self-annealed tracer molecules from the nonextractable fraction are filled-in with Tag polymerase and amplified, using the single stranded PCR adapter as a primer. The method has been applied to purification of fragments from a 2.9 kb plasmid added to E. coli DNA at equimolar quantity. Plasmid derived fragments (250-1000 bp), initially comprising 1/1400th part of tracer DNA, were purified to homogeneity after two rounds of subtraction followed by PCR.     

Securing urethral catheters in patients undergoing urethrovesical anastomosis

OBJECTIVE: Securing urethral catheters for the optimal healing of and prevention of extravasation at the urethrovesical anastomosis. METHOD: This article describes the technique that we use to help secure Foley catheters or replace them if necessary. CONCLUSION: In the rare patients where disruption of the anastomosis or loss of the catheter prematurely takes place, the nylon suture can be used to replace the catheter without jeopardizing the anastomotic line.     

Reliability of phase-velocity measurements of tibial bone

BACKGROUND AND PURPOSE: The Bone Stiffness Measurement Device-Swing is capable of measuring the propagation velocity of flexural waves in human tibial bone, which relates to bending stiffness. If the interrater and intrarater reliability of measurements obtained with the device are established, it can be used with confidence in assessing changes in bone. The purposes of this study were to detect potential sources of measurement error and to establish the interrater and intrarater reliability of measurements taken with the device. SUBJECTS AND METHODS: In the first part of the study, a random-effects design was used to obtain phase-velocity measurements in subjects without known orthopedic or neurological impairments. The second part of the study consisted of possible applications of the device with mixed designs on subjects with spinal cord injuries. By means of generalizability theory, multiple sources of error (eg, occasion, clinician, repetition) were estimated. For the clinical trial, 17 persons with spinal cord injuries not older than 5 weeks were tested. RESULTS: The standard error of measurement (SEM) for intrarater reliability measurements ranged from 7.3 to 9.8 m x s(-1) . The SEMs for interrater reliability measurements ranged from 5.7 to 9.5 m x s(-1). The SEMs for measurements obtained by a single clinician in a clinical population ranged from 11.9 to 39.7 m x s(-1). CONCLUSION AND DISCUSSION: The reproducibility of measurements obtained with the device is suitably high for the device to be used for evaluation in clinical and research settings.     

Effect of tylosin tartrate on humoral immune responses in chickens

This article examines the 50-year struggle of osteopathic physicians to gain entry into the US Military Medical Corps on the same basis as allopathic physicians. The author explores various arguments put forth by both physician groups, as well as those arguments presented by the respective surgeons general and relevant government officials.     

Ambulatory urodynamics

Ambulatory monitoring of lower and upper urinary tract continues to develop. The addition of electronic urine-loss measurements to indicate the exact time-related loss is an important new feature. In the near future, quantitative urine-loss measurements will become available. The additional possibility of plugging in a flowmeter completes the technique with respect to pressure-flow analysis. At present, however, ambulatory urodynamics is still confined to specialized urodynamic centers. Ambulatory urodynamic monitoring has shown that de-novo detrusor instability after a suspension operation is frequently missed as a preoperative diagnosis of detrusor overactivity. The first steps are being taken toward giving a quantitative analysis of detrusor activity during the filling phase. This justifies a more widespread use of this ambulatory monitoring.     

Expression of beta 1 integrins changes during transformation of avian lens epithelium to mesenchyme in collagen gels

Remarkably, a number of definitive epithelia, such as that of the anterior lens, give rise when suspended within 3D gels of type I collagen, to elongate, bipolar shaped cells that exhibit the ultrastructure, polarity, and migratory ability of mesenchymal cells. They begin producing type I collagen and stop producing crystallins, type IV collagen, and laminin. Here, we investigated changes in beta 1 integrins and their extracellular matrix (ECM) ligands during this transdifferentiation. The former free surface of the lens epithelium that is now in contact with collagen begins within a day to stain intensely for beta 1 and it is this surface rather than the surface facing the basement membrane that gives rise to mesenchymal cells. Immunoprecipitation experiments reveal a large increase in the beta 1 integrin subunit on mesenchymal cells as compared to the epithelium of origin. The alpha 5 integrin subunit, which is barely detectable in the lens, increases in the mesenchymal cells and alpha 3 continues to be expressed at about the same level as in the epithelium. alpha 6, the epithelial integrin subunit, and laminin, its ECM ligand, are not detected immunohistochemically or biochemically in the mesenchyme. Rather, the mesenchymal cells secrete abundant fibronectin, the major ECM ligand for alpha 5 beta 1. RGD peptides do not inhibit the transformation but antibodies to beta 1 do perturb the emigration of mesenchymal cells from the lens apical surface. We conclude that the beta 1 integrins newly expressed on the apical epithelial surface interact with the surrounding 3D collagen gel to help bring about this unusual epithelial-mesenchymal transition.