Reversible inactivation of purified leukocyte integrin CR3 (CD11b/CD18, alpha m beta 2) by removal of divalent cations from a cryptic site

Integrins exhibit reversible changes in their ability to bind ligands and these changes enable transient cell adhesion. We recently showed that leukocyte integrin CR3 (complement receptor type three, CD11b/CD18, alpha m beta 2) may be purified in a form that is either capable or incapable of binding soluble, monomeric ligand and that "inactive" CR3 may be rendered capable of binding ligand by addition of an anti-CR3 mAb known as KIM-127 (Cai and Wright, JBC. 270: 14358, 1995). Here, we demonstrate that active CR3 may be rendered inactive by treatment of immobilized receptor with EDTA. EDTA-treated CR3 failed to bind ligand even in the presence of mM Ca2+ and Mg2+, suggesting that EDTA-treatment caused a change in the receptor that is not readily reversed. EDTA-treated receptor did, however, bind ligand upon addition of KIM-127 plus Mg2+ with an affinity (17.8 +/- 4.5 nM) similar to untreated, active receptor (12.5 +/- 4.7 nM). EDTA-treated CR3 thus exhibits the properties of inactive CR3, in which the ligand binding site is cryptic but subject to exposure by KIM-127. A candidate for the cryptic ligand binding site is the I-domain, a Mg2+-binding region in the alpha chain of CR3. We found that monomeric C3bi binds directly to recombinant I-domain in a Mg(2+)-dependent fashion with an affinity of 300 +/- 113 nM. These results thus suggest that CR3 may be inactivated by removing tightly bound divalent cation from a cryptic site in CR3.     

Domain flexibility in retroviral proteases: structural implications for drug resistant mutations

Rigid body rotation of five domains and movements within their interfacial joints provide a rational context for understanding why HIV protease mutations that arise in drug resistant strains are often spatially removed from the drug or substrate binding sites. Domain motions associated with substrate binding in the retroviral HIV-1 and SIV proteases are identified and characterized. These motions are in addition to closure of the flaps and result from rotations of approximately 6-7 degrees at primarily hydrophobic interfaces. A crystal structure of unliganded SIV protease (incorporating the point mutation Ser 4 His to stabilize the protease against autolysis) was determined to 2.0 A resolution in a new space group, P3221. The structure is in the most "open" conformation of any retroviral protease so far examined, with six residues of the flaps disordered. Comparison of this and unliganded HIV structures, with their respective liganded structures by difference distance matrixes identifies five domains of the protease dimer that move as rigid bodies against one another: one terminal domain encompassing the N- and C-terminal beta sheet of the dimer, two core domains containing the catalytic aspartic acids, and two flap domains. The two core domains rotate toward each other on substrate binding, reshaping the binding pocket. We therefore show that, for enzymes, mutations at interdomain interfaces that favor the unliganded form of the target active site will increase the off-rate of the inhibitor, allowing the substrate greater access for catalysis. This offers a mechanism of resistance to competitive inhibitors, especially when the forward enzymatic reaction rate exceeds the rate of substrate dissociation.     

In vitro processing of herpes simplex virus type 1 DNA replication intermediates by the viral alkaline nuclease, UL12

Herpes simplex virus type 1 (HSV-1) DNA replication intermediates exist in a complex nonlinear structure that does not migrate into a pulsed-field gel. Genetic evidence suggests that the product of the UL12 gene, termed alkaline nuclease, plays a role in processing replication intermediates (R. Martinez, R. T. Sarisky, P. C. Weber, and S. K. Weller, J. Virol. 70:2075-2085, 1996). In this study we have tested the hypothesis that alkaline nuclease acts as a structure-specific resolvase. Cruciform structures generated with oligonucleotides were treated with purified alkaline nuclease; however, instead of being resolved into linear duplexes as would be expected of a resolvase activity, the artificial cruciforms were degraded. DNA replication intermediates were isolated from the well of a pulsed-field gel ("well DNA") and treated with purified HSV-1 alkaline nuclease. Although alkaline nuclease can degrade virion DNA to completion, digestion of well DNA results in a smaller-than-unit-length product that migrates as a heterogeneous smear; this product is resistant to further digestion by alkaline nuclease. The smaller-than-unit-length products are representative of the entire HSV genome, indicating that alkaline nuclease is not inhibited at specific sequences. To further probe the structure of replicating DNA, well DNA was treated with various known nucleases; our results indicate that replicating DNA apparently contains no accessible double-stranded ends but does contain nicks and gaps. Our data suggest that UL12 functions at nicks and gaps in replicating DNA to correctly repair or process the replicating genome into a form suitable for encapsidation.     

NK cells can recognize different forms of class I MHC

NK recognition and lysis of targets are mediated by activation receptor(s) whose effects may be over-ridden by inhibitory receptors recognizing class I MHC on the target. Incubation of normal lymphoblasts with a peptide that can bind to their class I MHC renders them sensitive to lysis by syngeneic NK cells. By binding to class I MHC, the peptide alters or masks the target structure recognized by an inhibitory NK receptor(s). This target structure is most likely an "empty" dimer of class I heavy chain and beta2m as opposed to a "full" class I trimer formed by binding of specific peptide that is recognized by CTL.     

Primary structure of gene H of cattle plague virus strain K]

The paper analyzes the standard legal and methodological assurance of the quality and safety of animal food raw materials and foodstuffs (meat, meat products, fish, shellfish, crayfish and their processing products) by the parasitic purity rates according the requirements under the Russian Federation's laws "On Sanitary and Epidemiological Well-Being of the Population", "On Protection of Consumer's Rights", "On Certification of Products and Services", those of SanPiN, such as 2.3.2.560-96 "Sanitary Requirements for the Quality and Safety of Food Raw Materials and Foodstuffs" and 3.2.569-96 "Prevention of Parasitic Diseases in the Russian Federation".     

Laser-speckle angular-displacement sensor: theoretical and experimental study

A novel, to our knowledge, method for the measurement of angular displacement for arbitrarily shaped objects is presented in which the angular displacement is perpendicular to the optical axis. The method is based on Fourier-transforming the scattered field from a single laser beam that illuminates the target. The angular distribution of the light field at the target is linearly mapped on a linear image sensor placed in the Fourier plane. Measuring this displacement facilitates the determination of the angular displacement of the target. It is demonstrated both theoretically and experimentally that the angular-displacement sensor is insensitive to object shape and target distance if the linear image sensor is placed in the Fourier plane. A straightforward procedure for positioning the image sensor in the Fourier plane is presented. Any transverse or longitudinal movement of the target will give rise to partial speckle decorrelation, but it will not affect the angular measurement. Furthermore, any change in the illuminating wavelength will not affect the angular measurements. Theoretically and experimentally it is shown that the method has a resolution of 0.3 mdeg ( approximately 5 murad) for small angular displacements, and methods for further improvement in resolution is discussed. No special surface treatment is required for surfaces giving rise to fully developed speckle. The effect of partially developed speckle is considered both theoretically and experimentally.     

Minimizing the average cost of paging and registration: A timer-based method

Methods of balancing call registration and paging are developed in this paper. Given that the probability distribution on the user location as a function of time is either known or can be calculated, previous work shows the existence of lower bounds on the average cost of paging. Here these bounds are used in conjunction with a Poisson incoming-call arrival model to formulate the paging/registration optimization problem in terms oftimeout parameters, _m ; the maximum amount of time to wait before registering given the last known location wasm. Timer-based methods, as opposed to location-based methods, do not require the user to record and process location information during the time between location updates. This feature might be desirable for minimizing mobile transceiver use during idle periods. We then consider uniform motion processes where a spatial translation of starting location produces an identical spatial translation of the associated time-varying probability distribution. This leads to a universal timeout parameter which may be readily calculated. We study and the minimum cost of paging/registration for a simple model of user motion and compare our results to an earlier method of location-based paging/registration cost minimization.     

A survey of problems,solution techniques,and future challenges in scheduling semiconductor manufacturing operations

In this paper, we discuss scheduling problems in semiconductor manufacturing. Starting from describing the manufacturing process, we identify typical scheduling problems found in semiconductor manufacturing systems. We describe batch scheduling problems, parallel machine scheduling problems, job shop scheduling problems, scheduling problems with auxiliary resources, multiple orders per job scheduling problems, and scheduling problems related to cluster tools. We also present important solution techniques that are used to solve these scheduling problems by means of specific examples, and report on known implementations. Finally, we summarize some of the challenges in scheduling semiconductor manufacturing operations.     

Sources and nature of secondary school teachers’ education in computer-related ergonomics

Teachers’ knowledge of computer-related ergonomics in education will have an effect on the learning process and the work practices of their students. However little is known about teacher education in this area. The study aimed to investigate the sources and nature of secondary school teachers’ education about computer-related ergonomics. It also investigated the use of computers by secondary school children. The study included all secondary schools in the Republic of Ireland. The findings suggest that computers are widely available and are used frequently by students and teachers. However, there is evidence that teachers are not receiving information on computer-related ergonomics. Teachers were generally dissatisfied with this and would welcome information on the subject. The study discusses the implications of the findings and makes recommendations to meet the needs of the teachers.     

Building language resources for a Multi-Engine English-Filipino machine translation system

In this paper, we present the building of various language resources for a multi-engine bi-directional English-Filipino Machine Translation (MT) system. Since linguistics information on Philippine languages are available, but as of yet, the focus has been on theoretical linguistics and little is done on the computational aspects of these languages, attempts are reported here on the manual construction of these language resources such as the grammar, lexicon, morphological information, and the corpora which were literally built from almost non-existent digital forms. Due to the inherent difficulties of manual construction, we also discuss our experiments on various technologies for automatic extraction of these resources to handle the intricacies of the Filipino language, designed with the intention of using them for the MT system. To implement the different MT engines and to ensure the improvement of translation quality, other language tools (such as the morphological analyzer and generator, and the part of speech tagger) were developed.