Transient gene expression from yeast artificial chromosome DNA in mammalian cells is enhanced by adenovirus

The introduction of high molecular weight DNA into mammalian cells is useful for gene expression studies. However, current transfection strategies are inefficient, necessitating propagation of stable DNA transformants prior to analysis of gene expression. Here we demonstrate that transient lipid-mediated DNA transfection can be used to assess gene expression from yeast artificial chromosomes (YACs) containing the 230 kb cystic fibrosis transmembrane conductance regulator gene ( CFTR ) and Escherichia coli lacZ . We also show that psoralen-UV inactivated adenovirus significantly enhances transfection efficiency. The ability to deliver high molecular weight DNA using lipid-mediated transfection should expedite the analysis of large human genes contained within artificial chromosome vectors.     

Effect of ozone therapy on hemostatic changes in patients with vascular atherosclerosis

Hemostatic changed induced by ozone therapy were studied in 81 patients with atherosclerosis of different vessels in 81 patients. It was found that use of ozone-oxygen mixtures leads to hypocoagulatory changes (diminution of platelet aggregation, lowering of fibrinogen concentration, prolongation of activated partial thromboplastin time, enhanced fibrinolytic activity) which contribute to clinical response.     

From phlebothrombosis to the postthrombotic syndrome

The same pathomechanism causes both thrombosis of the superficial and deep veins. For practical purposes one may separate phlebothrombosis of bed-patients and that of out-patients. Superficial phlebothrombosis and thrombophlebitis of the out-patients heal in shorter time by compression according to Heinrich Fisher. Patients should not be immobilized. Phlebothrombosis of the bed-patient should be reserved to clinical therapy including fibrinolysis, thrombectomy and anticoagulants. The post-thrombotic syndrome is treated usually by the practitioner. Treatment first has to activate drainage by lymphatic vessels and by paravenous tissue. Correct compression will improve circulation of fluid from tissue to capillaries. When balance is achieved, further physical measurements will be of help.     

Thermal beam distortions in end-pumped Nd:YAG, Nd:GSGG, and Nd:YLFrods

The thermally induced beam distortions in end-pumped Nd:YAG, Nd:GSGG, and Nd:YLF rods were analyzed and the influence of edge- and face-cooling was investigated. The distributions of temperature, stress, and strain in the crystals were calculated by finite element analysis. Based on these data, the space-resolved changes of the refractive index were determined considering thermal dispersion, surface deformation, and strain-induced birefringence. The resulting optical path difference for one round-trip in the end-pumped rods was integrated numerically. For each rod, the induced thermal lens was determined over the extent of the pump spot radius. The calculations of the optical path difference were experimentally confirmed by investigations using a modified Twyman-Green interferometer with a polarized HeNe probe beam at 633 nm under lasing and nonlasing conditions     

The University of Texas Health Science Center at San Antonio: experience with foot surgery in diabetics

The authors present their experience, at the University of Texas Health Science Center at San Antonio, with foot surgery in patients with diabetes mellitus. It is important to note that the results that follow are a reflection of the authors' experience with foot surgery in these patients from 1980 to 1985. The authors report two separate groups of patients: those who underwent elective foot surgery and those who underwent ablative foot surgery for an infected diabetic foot with necrosis. Also reviewed is the etiology of diabetic foot deformities and ulcerations. It has been the authors' experience that elective prophylactic surgery in patients with diabetes and an intact vascular status produces good results. Patients with an infected diabetic foot, providing they demonstrate adequate vascular status, have a high percentage of healing with early surgical intervention.     

The efficacy of radiofrequency lesioning of the cervical spinal dorsal root ganglion in a double blinded randomized study: no difference between 40 degrees C and 67 degrees C treatments

Marfan syndrome (MFS) is an autosomal dominant trait due to mutations in the fibrillin gene (FBN1). The MFS expressivity is variable, and its diagnosis relies completely on clinical criteria. Atypical cases and Marfan-like (marfanoid) clinical presentations are commonly found. The metacarpophalangeal pattern profile (MCPP), a radiological method in which the 19 tubular hand bones are assessed, has been used in the diagnosis of various syndromes. To investigate whether the MCPP was adequate to discriminate between MFS and Marfan-like subjects, we studied 38 patients who were referred to our service because they had an MFS diagnosis, diagnostic hypothesis, or differential diagnosis or had arachnodactyly with dolichostenomelia. Two groups were formed: 1) MFS: 21 patients with a mean age of 18.3 (10.8 S.D.) years and 2) Marfan-like syndromes: 16 patients who did not meet the current criteria, with a mean age of 14.6 (4.6 S.D.) years. The MCPP was performed in each case following the classical technique, and a characteristic mean profile was obtained for group I (MFS), with Z scores ranging from 0.69 to 2.73 (1.80+/-0.50; mean+/-S.D.). In group I, three cases had no correlation with the typical MFS pattern. In group II, three cases had an MFS pattern. The correlation with the mean MCPP of MFS permitted the differential diagnosis of MFS and marfanoid syndromes with 86% sensitivity, 81% specificity, and 86% positive and 81% negative predictive values. The results suggest that MCPP can be used effectively as an auxiliary tool in the nosology of these conditions and, because there is no change in MCPP with age, can be helpful in early diagnosis.     

Subcellular localization, substrate specificity and crystallization of duodenase, a potential activator of enteropeptidase

Duodenase, a serine protease from bovine duodenum mucosa, was located in endoplasmic reticulum, the Golgi secretory granules of epithelial cells and ducts of Brunner's glands by the A-gold immunocytochemical method. Duodenase exhibits trypsin-like and chymotrypsin-like specificities with a preference for substrates having lysine at the P1 and proline at the P2 positions. The kinetic constants for the hydrolysis of 21 potential duodenase substrates are reported. The best substrates were found to be alpha-N-tosylglycylprolyllysine 4-nitroanilide (k[cat]/Km of 35000 M[-1] s[-1]), alpha-N-succinylthreonylprolyllysine 4-nitroanilide (k[cat]/Km of 18000 M[-1] s[-1]) and alpha-N-serylprolyllysine 4-nitroanilide (k[cat]/Km of 2600 m[-1] s[-1]), all of which contain the P1-P3 sequence of the enteropeptidase zymogen/activation site. On the basis of its catalytic properties and sites of localization, duodenase has been postulated to be an activator of the enteropeptidase precursor. A tetradecapeptide (LVTQEVSPKIVGGS) having the P9-P5'sequence of the cleavage site of zymogen activation of bovine proenteropeptidase was synthesized, and kinetic parameters of its hydrolysis by duodenase were determined (Km of 87 microM; k[cat] of 1.4 s[-1]; k[cat]/Km of 16000 M[-1] s[-1]). Crystals of duodenase frozen in a stream of liquid nitrogen diffracted synchrotron X-rays to 0.2-nm resolution.