Temperature-regulated expression of the tac/lacl system for overproduction of a fungal xylanase in Escherichia coli

Temperature-regulated expression of recombinant proteins in the tac promoter (Ptac) system was investigated. Expression levels of fungal xylanase and cellulase from N. patriciarum in E. coli strains containing the natural lacI gene under the control of the Ptac markedly increased with increasing cultivation temperature in the absence of a chemical inducer. The specific activities (units per milligram protein of crude enzyme) of the fungal xylanase and cellulase produced from recombinant E. coli strain pop2136 grown at 42 degrees C were about 4.5 times higher than those of the cells grown at 23 degrees C and were even slightly higher when compared with cells grown in the presence of the inducer isopropyl beta-D-thiogalactopyranoside. The xylanase expression level in the temperature-regulated Ptac system was about 35% of total cellular protein. However, this system can not be applied to E. coli strains containing lacIq, which confers over production of the lac repressor, for high-level expression of recombinant proteins. In comparison with the lambda PL system, the Ptac-based xylanase plasmid in E. coli pop2136 gave a considerably higher specific activity of the xylanase than did the best lambda PL-based construct using the same thermal induction procedure. The high-level expression of the xylanase using the temperature-regulated Ptac system was also obtained in 10-litre fermentation studies using a fed-batch process. These results unambiguously demonstrated that the temperature-modulated Ptac system can be used for overproduction of some non-toxic recombinant proteins.     

Keratoacanthoma

The keratoacanthoma is a common cutaneous neoplasm that most often occurs on sun-exposed sites in light-skinned persons of middle age or older. It is considered the prototype of cutaneous pseudo-malignancies because it is a rapidly growing tumor with a histologic pattern resembling squamous cell carcinoma. It may be best viewed as an aborted malignancy that only rarely progresses into an invasive squamous cell carcinoma. It is most likely derived from hair follicle cells. The common type of keratoacanthoma and its many variants are discussed with emphasis on clinical and histologic features, biologic behavior, and response to therapy.     

The Mona Lisa identification: Evidence from a computer analysis

A recent computer-aided study identified the model immortalized in Leonardo da Vinci's celebratedMona Lisa to be none other than the artist himself. A follow-up investigation empolying similar techniques identifies the subject of asecond HiddenMona Lisa by the same artist. Analysis of photographic and x-ray images indicates that Leonardo first created a sketch of Isabella, Duchess of Aragon, which he later painted over with theMona Lisa, using himself as the model.     

A method for quantitative display of three-dimensional regional myocardial function

The DSR (dynamic spatial reconstructor), a multiple X-ray source scanner that generates stop action three-dimensional (3-D) images of a cylindrical volume, was used for quantitative imaging of left ventricular 3-D wall geometry and function in experimentally induced canine left ventricular myocardial infarction. Impaired regional myocardial function was induced by myocardial ischemia or infarction in four mongrel dogs by closed-chest occlusion of the proximal left anterior descending (LAD) coronary artery. At intervals of 6-14 weeks post occlusion, the dogs were scanned with the DSR during biatrial contrast injection. The 3-D shape, extent, and function of hypokinetic myocardium was measured from the DSR images utilizing measurement of the rate of local systolic wall thickening to detect regions of normal, ischemic, or scarred myocardium. The results were compared to scar size and anatomic distribution measured at postmortem examination. The anatomic extent and relationship of hypocontractile to normally contracting muscle was visualized by computer generated, pseudo 3-D shaded surface displays of the left ventricular chamber and by topographic projections of regional wall thickening rates onto a map of the left ventricular endocardial surface. The location of myocardial infarction and the surrounding zone of impaired function is clearly defined by this 3-D CT scanning procedure. The display method presented here provides both localization and quantification of the volume of ischemic and infarcted myocardium.     

Electrodeposition of Fe−Ni−Co alloys part I: Direct current deposition

Effects of hydrodynamic conditions, current density and solution temperature on the d.c. electrodeposition of Fe–Ni–Co alloys have been investigated with stationary planar and rotating cylindrical electrodes. The deposition rate of Fe showed mass transfer effects at cathodic potentials –1.35 V/sce. Deposition of Ni appeared to be kinetically controlled; deposition of Co appeared to be under kinetic control at potentials –1.35 V/sce but under mixed control at –1.65 V. Current efficiency of the codeposition process increased with increasing current density and decreased with increasing rotation rate. Higher solution temperatures and rotation rates extended the applied current density range where smooth, adherent, and metallic-looking deposits could be obtained. An increase in solution temperatures also decreased anomalous codeposition of Fe–Ni–Co. Calculations based on the Hessami-Tobias model provide qualitative agreement with dependence of experimental electrodeposition on applied current density, hydrodynamics and temperature.     

Effect of specific dietary fatty acids on lipogenesis in the livers and mammary glands of lactating mice

The effects of linoleic, linolenic and columbinic acids fed as 4% of a high carbohydrate (50% glucose) diet on the activities and the amounts of several enzymes associated with fatty acid synthesis in livers and mammary glands of lactating mice were compared with those for stearic and oleic acids. Fatty acid synthesis, measured in vivo, was significantly lower in livers of mice ingesting all 3 polyunsaturated fatty acids (PUFA), whereas in mammary glands synthesis was lower only in mice receiving columbinic acid. The activities of fatty acid synthetase (FAS) and acetyl CoA carboxylase were significantly reduced in liver by all 3 PUFA, as were activities of glucose-6-phosphate dehydrogenase, malic enzyme (ME) and citrate cleavage enzyme (CCE), also associated with lipogenesis. In mammary gland, on the other hand, the activities of these enzymes were unaffected by dietary PUFA. The tissue contents of FAS, ME and CCE, measured by rocket immunoelectrophoresis, were found to be significantly reduced in liver by linoleate, linolenate and columbinate but were not significantly altered in mammary gland. The decrease in hepatic lipogenesis observed was principally due to a decrease in the amounts of these enzymes induced by the dietary PUFA but the inhibition in mammary gland caused by columbinate could not be accounted for by a reduction in enzyme contents and therefore may be due to allosteric effects which occur when fatty acid synthesis is measured with³H₂O. The fatty acid composition in liver and mammary gland of dams and in liver and kidney of pups completely reflected dietary fatty acids. Columbinate made up ca. 20% of the total fatty acids in both tissues of the columbinic acid-fed mice and ca. 15% in the pup tissues. This suggests that columbinate is incorporated into milk lipids of dams and is easily absorbed by pups. The elevated ratios of 16/16∶1 and 18/18∶1 in liver and mammary gland of dams and liver and kidney of the pups from dams fed linoleate, linolenate and columbinate suggest that each of these polyunsaturated fatty acids in the diet can inhibit the activity of Δ9 desaturase.     

Archival Appraisal and the Digital Record: Applying Past Tradition for Future Practice

The digital revolution has changed the daily work of archivists. As a result, archivists have refined their analog practices and initiated new ones to accommodate the unique information creation and access needs of users. While limitless storage and technological obsolescence may be surmountable challenges, appraisal continues to be a necessary component of this preservation enterprise. Macro-appraisal—a top-down functional assessment of an organization’s high-level decision making that defines the enduring value of specific bodies of documentary evidence of these creative or administrative processes—is used to facilitate the management of diverse bodies of born-digital records in archives and special collections.     

Loss of heterozygosity and overexpression of the p53 gene in ovarian carcinoma

Eight anticonvulsant drugs-including clonazepam, diazepam and phenobarbital-were tested for their effects on GABA-stimulated chloride uptake in rat cerebral cortical microsacs (unfiltered synaptoneurosomes). "Mid" and "high" therapeutic concentrations were screened, and, if significant enhancement was found, full concentration-response tests were done. In the initial screens, enhancement of GABA-stimulated uptake was found only with phenobarbital, clonazepam and diazepam. In subsequent concentration-response tests, the effects of phenobarbital were found to occur throughout the range of normal, anticonvulsant concentrations, whereas the effects of clonazepam and diazepam were observed only above the concentrations normally used for the chronic control of seizures or anxiety. These data suggest that phenobarbital's anticonvulsant effects are mediated via the GABAA receptor complex, but that the low-dose effects of the benzodiazepines may be mediated via some other mechanism.