Crystal structure of tRNA-guanine transglycosylase: RNA modification by base exchange

tRNA-guanine transglycosylases (TGT) are enzymes involved in the modification of the anticodon of tRNAs specific for Asn, Asp, His and Tyr, leading to the replacement of guanine-34 at the wobble position by the hypermodified base queuine. In prokaryotes TGT catalyzes the exchange of guanine-34 with the queuine (.)precursor 7-aminomethyl-7-deazaguanine (preQ1). The crystal structure of TGT from Zymomonas mobilis was solved by multiple isomorphous replacement and refined to a crystallographic R-factor of 19% at 1.85 angstrom resolution. The structure consists of an irregular (beta/alpha)8-barrel with a tightly attached C-terminal zinc-containing subdomain. The packing of the subdomain against the barrel is mediated by an alpha-helix, located close to the C-terminus, which displaces the eighth helix of the barrel. The structure of TGT in complex with preQ1 suggests a binding mode for tRNA where the phosphate backbone interacts with the zinc subdomain and the U33G34U35 sequence is recognized by the barrel. This model for tRNA binding is consistent with a base exchange mechanism involving a covalent tRNA-enzyme intermediate. This structure is the first example of a (beta/alpha)-barrel protein interacting specifically with a nucleic acid.     

193-nm lithography

The trend in microelectronics toward printing features 0.25 μm and below has motivated the development of lithography at the 193-nm wavelength of argon fluoride excimer lasers. This technology is in its early stages, but a picture is emerging of its strengths and limitations. The change in wavelength from 248 to 193 nm will require parallel progress in projection systems, optical materials, and photo-resist chemistries and processes. This paper reviews the current status of these various topics as they have been engineered under a multiyear program at MIT Lincoln Laboratory     

The natural killer cell receptor specific for HLA-A allotypes: a novel member of the p58/p70 family of inhibitory receptors that is characterized by three immunoglobulin-like domains and is expressed as a 140-kD disulphide-linked dimer

Human natural killer (NK) cells express inhibitory receptors that are specific for different groups of HLA-C or HLA-B alleles. The majority of these receptors belong to the immunoglobulin (Ig) superfamily and are characterized by two or three extracellular Ig-like domains. Here we describe a novel inhibitory NK receptor that is specific for a group of HLA-A alleles. The HLA-A3-specific NK cell clone DP7 has been used for mice immunization. Two mAbs, termed Q66 and Q241, bound to the immunizing clone and stained only a subset of NK cell populations or clones. Among Q66 mAb-reactive clones, we further selected those that did not express any of the previously identified HLA-class I-specific NK receptors. These clones did not lyse HLA-A3+ (or -A11+) target cells, but lysis of these targets could be detected in the presence of Q66 or Q241 mAbs. On the other hand, target cells expressing other HLA-A alleles, including -A1, -A2, and -A24, were efficiently lysed. Moreover, none of the HLA-C or HLA-B alleles that were tested exerted a protective effect. Q66+, but not Q66- NK cell clones, expressed messenger RNA coding for a novel 3 Ig domain protein homologous to the HLA-C (p58) and HLA-B (p70) receptors. The corresponding cDNA (cl.1.1) was used to generate transient and stable transfectants in COS7 and NIH3T3 cell lines, respectively. Both types of transfectants were specifically stained by Q66 and Q241 mAbs. Since the cytoplasmic tail of Q66-reactive molecules was at least 11 amino acid longer than the other known p58/p70 molecules, we could generate an antiserum specific for the COOH-terminus of Q66-reactive molecules, termed PGP-3. PGP-3 immunoprecipitated, only from Q66+ NK cells, molecules displaying a molecular mass of 140 kD, under nonreducing conditions, which resolved, under reducing conditions, in a 70-kD band. Thus, differently from the other p58/p70 receptors, Q66-reactive molecules appear to be expressed as disulphide-linked dimers and were thus termed p140. The comparative analysis of the amino acid sequences of p58, p70, and p140 molecules revealed the existence of two cysteins proximal to the transmembrane region, only in the amino acid sequence of p140 molecules.     

Autonomous vehicles

There are various kinds of autonomous vehicles (AV's) which can operate with varying levels of autonomy. This paper is concerned with underwater, ground and aerial vehicles operating in a fully autonomous (nonteleoperated) mode. Further this paper deals with AV's as a special kind of device, rather than full-scale manned vehicles operating unmanned. The distinction is one in which the AV is likely to be designed for autonomous operation rather than being adapted for it as would be the case for manned vehicles. We provide a survey of the technological progress that has been made in AV's, the current research issues and approaches that are continuing that progress, and the applications which motivate this work. It should be noted that issues of control are pervasive regardless of the kind of AV being considered, but that there are special considerations in the design and operation of AV's depending on whether the focus is on vehicles underwater on the ground, or in the air. We have separated the discussion into sections treating each of these categories     

Cyclobutanones as markers for irradiated samples of Brie and Camembert cheese

The feasibility of applying a chemical method based on the formation of 2-alkylcyclobutanone to samples of irradiated soft cheese (Brie and Camembert) was investigated. Significant quantities of 2-dodecylcyclobutanone (DCB) and 2-tetradecylcyclobutanone (TCB) were detected in both types of irradiated cheese and are proposed as qualitative markers. Other members of the cyclobutanone family (decyl- and tetradecenyl-) are also thought to be present but could not be substantiated due to a lack of authentic standards. These compounds were absent from the unirradiated samples. Results also show a significant linear relationship between the irradiation dose (1 to 8 kGy) and the amount of DCB and TCB detected in the cheese.     

Improved accuracy of burn wound assessment using laser Doppler

The utility of the laser Doppler for determining burn depth has been questioned because of problems with technology and methodology. This study prospectively evaluates the ability of a new laser Doppler technique to predict burn healing time. Using the Periflux System 4000 laser Doppler, readings were taken on 305 burns (147 patients) on postburn day 3 or 4. Sixty-six wounds were used to derive a predictive function (phase I) and 152 wounds were used to test the function (phase II). Blood flow dynamics (flux), microvascular dilation capacity of the wounds to beat stress, and flow motion wave pattern (vasomotion) were studied using the laser Doppler, and seven parameters were evaluated to determine their relative contribution to the prediction of healing time. These parameters are hyperemic flux (flux value after heating to 42 degrees C), average hyperemic wave amplitude (AHWA), number of average flux units >100(F100), number readings with wave amplitude 75 (A5), average flux change (AFC), percentage of average flux increase, and relative flow capacity (RFC = AFC/average hyperemic flux). After readings were made, the wounds were observed and divided into two groups: those that healed in less than 14 days and those that healed or were grafted after 14 days. A step-wise discriminant analysis was used to assess the relative contribution of the Doppler-derived measures to healing time prediction. AHWA, F100, and RFC were included in the final discriminant function explaining 72% of the healing time variance (Wilks' lambda value 0.28; p value <0.0001). Predicted outcome = 0.05(AHWA) + 0.31(F100) + 5.0(RFC) - 2.3. With this derived function, there is 94% accuracy in the prediction of burn wound healing time compared with a physician predictive accuracy of 70%.     

Cerebral protection by intermittent reperfusion during temporary focal ischemia in the rat

Temporary arterial occlusion has been routinely used as an adjunct in intracranial aneurysm surgery. This has commonly been performed using a protocol of multiple short periods of occlusion alternating with periods of restoration of normal circulation. Recently, the logical basis of this method has come under scrutiny. There is extensive experimental evidence to suggest that repetitive, brief periods of global ischemia may cause more severe cerebral injury than an equivalent single period of global ischemia. Only recently has this issue begun to be addressed with regard to focal ischemia. Hence, despite the common use of temporary clipping, little experimental data are available regarding the ischemic consequences of temporary arterial occlusion with periods of reperfusion versus uninterrupted temporary occlusion. To investigate this issue, a protocol of occlusion/reperfusion that simulates the temporal profile that occurs during surgery was performed in a rat model of focal ischemia. Sixteen anesthetized Sprague-Dawley rats were divided into two groups. The animals in Group I underwent 60 minutes of uninterrupted middle cerebral artery occlusion and the animals in Group II were subjected to six separate 10-minute occlusion periods with 5 minutes of reperfusion between occlusions. Histopathological analysis was performed 72 hours postischemia. Group I had significantly increased mean infarction volumes (50.0 +/- 12.1 mm3) compared to Group II (8.7 +/- 3.1 mm3) (p = 0.008). Injuries in Group I occurred in both the cortex and striatum, whereas Group II showed only striatal injuries. Furthermore, the extent of the injuries in Group II was less severe, characterized by ischemic neuronal injury rather than frank infarction. The results indicate that intermittent reperfusion is neuroprotective during temporary focal ischemia and support the hypothesis that intermittent reperfusion is beneficial if temporary clipping is required during aneurysm repair.     

Room temperature characterization of Hg1-xCdxTe P-on-n heterostructure photodiodes

Measurements of 77K R_oA and 300K reverse bias dynamic impedance (R_dA) products at one volt reverse bias has been carried out to assess the degree of correlation of this figure of merit. Planar P-on-n heterostructures were grown on near lattice-matched CdZnTe substrates with Hg_1-xCd_xTe (0.20< x <0.30) by molecular beam epitaxy. These devices were passivated with CdTe and doped with indium and arsenic as n- and p-type dopants, respectively. Current-voltage characteristic of these devices exhibit thermally generated dark currents at small and modest reverse bias. We have observed that R_oA values of these long wavelength infrared P-on-n heterostructure photodiodes at 77K correlate with room temperature R_dA values. Diode arrays with high room temperature R_dA values at one volt reverse bias also have high R_oA values at 77K. Similarly, low R_dA values at room temperature indicate poor performance at 77K where deviation from diffusion current occurs at reverse bias of 0.2 to 1 volt at room temperature. The results presented here, for a small samples of devices, demonstrate that room temperature measurements of current-voltage characteristics to evaluate Hg_1-xCd_xTe (0.22< x <0.28) diode performance and array uniformity at lower temperatures can be used. This provides an acceptable criteria for further study at lower temperatures.     

AC measurements of the minimum longitudinal resistance of a QHEsample from 10 Hz to 10 kHz

AC measurements of the longitudinal resistance, R_xx, of a quantum Hall effect (QHE) sample have been made in a frequency range from 10 Hz to 10 kHz. The results show no frequency effect on the minimum value of R_xx corresponding to the quantum numbers i=2 and i=4, within the measurement resolution of 0.5 mΩ. Therefore, the influence of frequency on the value of the quantized Hall resistance, R_H, should not exceed a few parts in 10⁹ . Some unwanted effects detected during the development of the resistance bridge have been pointed out