Deuterium off-resonance rotating frame spin-lattice relaxation of macromolecular bound ligands

Deuterated 3-trimethylsilylpropionic acid binding to bovine serum albumin was used as a model system to examine the feasibility and limitations of using the deuterium off-resonance rotating frame spin-lattice relaxation experiment for the study of equilibrium ligand-binding behavior to proteins. The results of this study demonstrate that the rotational-diffusion behavior of the bound species can be monitored directly, i.e., the observed correlation time of the ligand in the presence of a protein is approximately equal to the correlation time of the ligand in the bound state, provided that the fraction of bound ligand is at least 0.20. The presence of local ligand motion and/or chemical exchange contributions to relaxation in the bound state was inferred from the observation that the correlation time of the bound ligand was somewhat smaller than the correlation time characterizing the overall tumbling of the protein. An approximate value for the fraction of bound ligand was obtained from off-resonance relaxation experiments when supplemental spin-lattice or transverse relaxation times were employed in the analysis. Incorporation of local motion effects for the bound species into the theoretical relaxation formalism enabled the evaluation of an order parameter and an effective correlation time, which in conjunction with a wobbling in a cone model, provided additional information about ligand motion in the bound state.     

Localization of prothymosin alpha in the nucleus

Age differences in processing resources seem salient to age-related declines in secondary (or "recent") memory. Community-dwelling adults (N = 90, ages 30-80) completed 4 memory tests: Wechsler Memory Scale-Revised (WMS-R) Logical Memory (LM), Cowboy Story (CS), WMS-R Visual Reproductions (VR), and Extended Complex Figure Test (ECFT; Fastenau, in press). Two space-capacity measures (WMS-R Digit Span and Visual Memory Span) and 4 processing speed measures (cancellation and mental-tracking tasks) assessed processing resources. A statistical control procedure was used to isolate retrieval efficiency and measures contributions of age and processing resources to retrieval. A negative relationship between age and retrieval efficiency emerged on all measures (p < .05). The age effect was reduced 60% on LM and CS when processing resources were controlled, eliminated for VR, and unchanged on ECFT. It is possible that visual-spatial retrieval requires fewer processing resources than does verbal retrieval.     

Promoter elements controlling developmental and environmental regulation of a tobacco ribosomal protein gene L34

The rpL34 gene, which encodes a cytoplasmic ribosomal protein with a high homology to the rat 60S r-protein L34, was isolated from a genomic library of tobacco (Nicotiana tabacum L. cv. Xanthi-nc). A 1500 bp upstream promoter fragment was fused to the chloramphenicol acetyltransferase (CAT) reporter gene or beta-glucuronidase (GUS) reporter gene and transferred into tobacco plants by the Agrobhacterium-mediated leaf disk transformation method. Analysis of CAT activity in leaf tissues showed that mechanical wounding increased the rpL34 promoter activity about 5 times as compared to untreated controls and that the promoter activity was further enhanced by plant growth regulators, 2,4-dichlorophenoxyacetic acid and benzyladenine. Histochemical GUS staining patterns of the transgenic plants showed that the rpL34 promoter activity is high in actively growing tissues, including various meristems, floral organs, and developing fruits. A series of 5' deletion analyses of the rpL34 promoter indicated that a 50 bp region located between -179 and -129 is essential for wound, auxin and cytokinin responses. Deletion of this region reduced the promoter activity to an undetectable level. Insertion of the 50 nucleotide sequence into a minimal promoter restored the promoter activity and the promoter strength was proportional to the copy number of the upstream sequence. The role of TATA and CAAT box regions was studied by a series of 3' deletion analyses. A 3' deletion up to -28 did not significantly affect the promoter strength. However deletion of the promoter up to 70 bp, which deleted the TATA box region, significantly reduced promoter activity. Further deletion of the promoter up to - 104. eliminating the CAAT box region, abolished the promoter activity. These results suggest that the TATA box and CAAT box regions are also important for the rpL34 promoter activity in addition to the 50 bp upstream region.     

Management of choledocholithiasis in patients undergoing laparoscopic cholecystectomy

The development of laparoscopic cholecystectomy has rekindled the issue of management of choledocholithiasis. A number of options exist including pre or postoperative endoscopic sphincterotomy (ERCP-ES), laparoscopic common duct exploration or open common duct exploration. We present here our experience with the management of choledocholithiasis in patients treated with laparoscopic cholecystectomy. From January 1991 to January 1995, 900 patients underwent laparoscopic cholecystectomy. 71 ERCP were carried out in 71 patients with suspicion or evidence of choledocholithiasis. Common duct stones were detected in 44 patients. Preoperative ERCP was done in 56 patients, with suspicion of choledocholithiasis, based on clinical, laboratory or ultrasonographic findings. 29 of these patients (51.7%) had common duct stones, that were successfully removed by endoscopic sphincterotomy. One patient suffered mild pancreatitis and a second one had transient hyperamylasemia. Postoperative ERCP was performed in 15 patients. Indications for ERCP were the evidence of common duct stones in intraoperative cholangiography in 7 cases, and clinical or laboratory suspicion of choledocholithiasis, 3 months to 3 years after laparoscopic cholecystectomy. Stones were detected in 100% of the patients. In 11 patients (73.3%), the stones were extracted by endoscopic sphincterotomy and 4 patients underwent open common duct exploration. Two patients had transient hyperamylasemia. ERCP is a safe and effective method for detection and treatment of common duct stones. ERCP prior to laparoscopic cholecystectomy in patients suspected of having choledocholithiasis, is safe and offers with good results. Rutinary intraoperative cholangiography is recommended, for the detection of unsuspected choledocholithiasis and as an effective treatment (postoperative-ERCP, open or laparoscopic common duct exploration) can be chosen depending on surgeon's skills and patient's characteristics.     

Ultrasound biomicroscopy of the anterior segment

BACKGROUND: New imaging technologies are revolutionizing the understanding and treatment of a wide variety of ocular disorders. Confocal scanning laser ophthalmoscopy, ultrasound biomicroscopy, confocal scanning laser polarimetry, color doppler imaging of blood flow, and optical coherence tomography are providing important information regarding disease pathophysiology, diagnosis, progression, and treatment. METHODS: High frequency (50 MHz), high resolution ultrasound biomicroscopy of the anterior segment was obtained in a wide variety of disorders of the anterior segment. Tissue resolution is approximately 50 microns and the penetration depth is 5 mm. RESULTS: Ultrasound biomicroscopy is capable of imaging the comea, iris, anterior chamber, anterior chamber angle, posterior chamber, and ciliary body with great detail. The structures surrounding the posterior chamber, previously hidden from clinical observation, can be imaged and their normal anatomic relationships assessed. The various forms of angle closure glaucoma, such as pupillary block and plateau iris configuration, can be differentiated. The concave iris found in pigment dispersion and its response to treatment can be assessed. Visualization of anterior segment anatomy in eyes with opaque media is possible. CONCLUSIONS: Ultrasound biomicroscopy assists in the management of eyes with disorders of the anterior segment. Future applications of this technology will yield important information regarding accommodation, normal ocular physiology and disease pathophysiology.     

Elevated levels of basic fibroblast growth factor in patients with limb ischemia

Basic fibroblast growth factor (bFGF), a prototypic member of a family of heparin-binding growth factors, is angiogenic both in vitro and in vivo. Increased levels and activity of bFGF have been documented in a variety of diseases, including tumors. We sought to determine whether bFGF might be similarly elevated in patients with clinical evidence of limb ischemia. Serum was obtained at the time of percutaneous revascularization from patients with symptomatic peripheral vascular disease (46 procedures were performed on 40 patients). An enzyme-linked immunoassay specific for bFGF was used (limit of detection, 1 pg/ml; range in normal subjects, 0 to 5 pg/ml). Among the 40 patients (28 men, 12 women, mean age 70 years) studied, elevated circulating bFGF (> or = 10 pg/ml) was detected in 36 samples (78%); levels ranged from 10 to 310 pg/ml (mean +/- SEM = 62 +/- 12). In 16 (89%) of 18 patients with both rest pain and nonhealing ischemic ulcers, serum bFGF levels were elevated up to 30 times normal values. In conclusion, circulating levels of bFGF are elevated in patients with vascular insufficiency and may reflect a physiologic response to limb ischemia.     

3H]-SR 27897B: a selective probe for autoradiographic labelling of CCK-A receptors in the brain

The binding and distribution of radiolabelled SR27897B, a potent CCK-A antagonist, was characterized using in vitro receptor autoradiography. Rapid imaging and quantitative analysis of [3H]SR27897B binding was obtained in a very short period of time (5 days) with a highly sensitive radioimager ensuring very short exposure times for isotopes such as tritium. Tritiated SR27897B binding sites are localized almost exclusively in the area postrema and the medical part of the nucleus tractus solitarius and in this nucleus the rostral-caudal distribution of CCK-A sites differed from that of sulphated CCK8 receptors. Receptor binding properties analyzed on 15 microns serial coronal sections showed on site receptor occupancy in these two regions with high affinity and selectivity characteristic of the CCK-A receptor. These results precisely locate the SR27897B binding sites and provide further support for the absence of heterogeneity of the CCK-A receptors in the rat brain.     

Prednisolone therapy of idiopathic feline lower urinary tract disease: a double-blind clinical study

A double-blind clinical study was performed to evaluate prednisolone as treatment for idiopathic feline lower urinary tract disease. No differences in response were observed in prednisolone- and placebo-treated cats.