Insertional re-activation of a chloramphenicol acetyltransferase misfolding mutant protein

The deletion of nine residues from the C-terminus of the bacterialchloramphenicol acetyltransferase (CAT) results in depositionof the mutant protein in cytoplasmic inclusion bodies and lossof chloramphenicol resistance in Escherichia coli. This foldingdefect is relieved by C-terminal fusion of the polypeptide withas few as two residues. Based on these observations, efficientpositive selection for the cloning of DNA fragments has beendemonstrated. The cloning vector encodes a C-terminally truncatedCAT protein. Restriction sites in front of the stop codon allowthe insertion of target DNA, resulting in the production ofproperly folded CAT fusion proteins and regained chloramphenicolresistance. The positive selection of recombinants is accomplishedby growth of transformants on chloramphenicol-containing agarplates. The method appears particularly convenient for the cloningof DNA fragments amplified by the PCR because minimal informationto restore CAT folding can be included in the primers. The cloningof random sequences shows that the folding defect can be relievedby fusion to a wide variety of peptides, providing great flexibilityto the positive selection system. This vector may also contributeto the determination of the role of the C-terminus in CAT folding.     

Unexplained physical symptoms: outcome, utilization of medical care and associated factors

BACKGROUND: Measurement of intracardiac hemodynamic parameters has been limited to brief periods in the acute care setting. We developed and evaluated an implantable hemodynamic monitor that is capable of measuring chronic right ventricular oxygen saturation and pulmonary artery pressure. METHODS AND RESULTS: The device consists of an electronic controller placed subcutaneously and two transvenous leads placed in the right ventricle (reflectance oximeter) and pulmonary artery (variable capacitance pressure sensor). Implantation was performed in 10 patients with severe left ventricular dysfunction. Average implant pulmonary artery pressures were systolic, 52 +/- 16 mm Hg; diastolic, 29 +/- 11 mm Hg; and mean, 40 +/- 12 mm Hg. The mean right ventricular oxygen saturation at implant was 51%. Provocative maneuvers, including postural changes, sublingual nitroglycerin, and bicycle exercise, demonstrated expected changes in measured oxygen saturation and pulmonary artery pressures over time. At follow-up of 0.5 to 15.5 months, there were no significant differences between pulmonary artery pressures or oxygen saturation values transmitted from the device and simultaneous measurement with balloon flotation catheters. Four of the pulmonary artery leads dislodged and three demonstrated sensor drift, whereas two of the oxygen saturation sensors failed. Four patients died and four received transplants. Pathological study did not demonstrate injury to the right ventricular outflow tract or pulmonic valve. CONCLUSIONS: Chronic measurement of hemodynamic parameters in the outpatient setting with implantable sensor technology appears to be feasible. The devices are well tolerated without significant untoward effects, and the sensors generally function well over time, providing reliable information. Clinical usefulness remains to be established.     

Duration of spinal anaesthesia is determined by the partition coefficient of local anaesthetic

We have compared the duration of motor block produced by four local anaesthetics administered into a chronically implanted subarachnoid catheter in rabbits. Each group (n = 6) received four different doses of amethocaine, bupivacaine, lignocaine or procaine, and the duration of the resulting motor block was assessed. Dose-response curves were plotted for each drug. As a measure of activity of the anaesthetics, we used the dose of each drug required to produce block of 60-min duration (D60 min) and the correlation between D60 min and different drug properties was examined. An inverse linear correlation (r = 0.995; P < 0.01) was observed between log D60 min and the log of the partition coefficient of the local anaesthetics. No correlation was found between the effect and degree of protein binding, pKa or molecular weight. These results suggest that, in spinal anaesthesia, the partition coefficient could be used as a predictor of the duration of anaesthetic action.     

Quality of Crude Fish Oil Extracted from Herring Byproducts of Varying States of Freshness

ABSTRACT: Herring byproducts were stored at 2 and 15 °C for up to 72 h. Over time, significant increases of total volatile bases (TVB), histamine, putrescine, cadaverine, and tyramine were detected. However, only tyramine and TVB levels were temperature-dependent. The level of total polyunsaturated fatty acids (PUFAs) was constant. Longer byproducts storage gave rise to an oil with higher levels of free fatty acids, eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and total PUFAs, while fluorescent compounds were lower. A higher storage temperature led to oil higher in α-tocopherol and EPA levels and lower in anisidine value. Surprisingly, the oil with the highest content of PUFAs was not produced from the freshest byproducts, and oil with low oxidation products can be extracted from stored byproducts.     

Effect of orienting fields on agglomeration in dilute dispersions of magnetic fine particles

Static light scattering was used to investigate dilute dispersions of fine gamma -Fe/sub 2/O/sub 3/ magnetic particles. The particles were acicular and on average 0.39 mu m long and 0.056 mu m in diameter. They were dispersed in organic solvents at concentrations of approximately 10/sup -4/ vol%, corresponding to an average interparticle separation of 9 mu m. Computer programs using a coupled dipole algorithm to predict the scattering from the particles were written. Experimental results do not agree with computer calculations of the light scattering from single particles, but are consistent with scattering from thick ropelike agglomerates of particles. The size of these agglomerates was estimated after 10 s to be less than 100 particles, while the thickness of the ropes is at least 5 particles.<>     

Hydrogen adsorption studies in micro-size cobalt dots

Hydrogen desorption curves were obtained from a sample composed of a square arrangement of Co dots with average diameter of 4.4 μm, separated by a distance of 11.6 μm. A macroscopic sample of Co dots grown on a 2.5 × 2.5 cm Si substrate was made by standard lithographic techniques and used in these experiments. Thermal programmed desorption (TPD) was performed under ultra-high vacuum conditions. Hydrogen TPD curves were obtained from a 1 × 1 cm Co dots samples displaying a maximum of intensity at 425 K. Hydrogen TPD curve was also obtained from 1 cm× 1 cm samples of Co films and Co foils for comparison. The hydrogen TPD curves have decreasing intensity from the Co foils to the Co dots and finally to the Co films. This indicates that there are more sites for hydrogen adsorption on the Co dots than in the Co films. This is a surprising result because there is approximately 8.7 times less Co atoms exposed in the Co dots that in the Co film sample. A desorption energy of 27 kcal/mol was obtained for the Co dots suggesting that hydrogen is adsorbed on an hcp hollow site of the Co dot crystalline structure.     

The hypersonic environment: Required operating conditions and design challenges

Hypersonic flight powered by airbreathing engines offers the potential for faster response time at long ranges, and reduced cost for access-to-space. In the present paper the operating environment of typical hypersonic vehicles are discussed, including results for the radiation equilibrium wall temperature of external vehicle surfaces and the flow properties through three sample engines spanning the range of hydrocarbon-fueled Mach 4-8 flight and hydrogen-fueled flight at speeds up to Mach 17. Flow conditions at several locations through the sample engines were calculated to provide indications of the required operating flow environment. Additional system consideration such a seals, joints, vehicle integration and in-service engineering are addressed.     

From biological inspiration toward next-generation manipulators: manipulator control focused on human tasks

This paper presents our approach to extend the niche of behavior-based robotics toward manipulation. We use results from neuroscience to derive some qualitative design rules for the mechanics of the manipulator, resulting in a next-generation manipulator, the "soft arm". By defining the basic behaviors of the manipulator as trajectory-producing behaviors (which is also biologically plausible), we have designed a first test case: writing on a board with a mobile manipulator. The soft arm has not yet been developed; therefore, we have emulated such a soft robot arm on an industrial robot.     

Validity of the Parabolic Effective Mass Approximation in Silicon and Germanium n-MOSFETs With Different Crystal Orientations

This paper investigates the validity of the parabolic effective mass approximation (EMA), which is almost universally used to describe the size and bias-induced quantization in n-MOSFETs. In particular, we compare the EMA results with a full-band quantization approach based on the linear combination of bulk bands (LCBB) and study the most relevant quantities for the modeling of the mobility and of the on-current of the devices, namely, the minima of the 2-D subbands, the transport masses, and the electron density of states. Our study deals with both silicon and germanium n-MOSFETs with different crystal orientations and shows that, in most cases, the validity of the EMA is quite satisfactory. The LCBB approach is then used to calculate the values of the effective masses that help improve the EMA accuracy. There are crystal orientations, however, where the 2-D energy dispersion obtained by the LCBB method exhibits features that are difficult to reproduce with the EMA model.