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Many inference algorithms develop the time-domain connection of the direct by employ a Kalman filter base on a first-order (or sometimes second-order) estimate of the time-varying channel amid a norm based on link toning (CM), or on the Minimization of Asymptotic Variance (MAV). To decrease the complexity of the high-dimensional RW-KF for combined opinion of the multi-path multifaceted amplitudes, we suggest using an inferior dimensional RW-KF that estimate the compound amplitude of each path separately. We demonstrate that this amounts to a simplification of the joint multi-path Kalman increase formulation through the Woodbury’s identities. Hence, this innovative algorithm consists of a superposition of self-determining single-path single-carrier KFs, which be optimized in our earlier studies. This examination allow us to settle in the optimization to the authentic multi-path multi-carrier scenario, to afford logical formulas for the mean-square error presentation and the best tuning of the future estimator in a straight line as a function of the physical parameter of the canal (Doppler frequency, signal-to-noise-ratio, power delay profile). These logical formula are known for the first-, second-, and third-order RW models used in the KF. The future per-path KF is exposed to be as well-organized as the accurate KF (i.e., the joint multi-path KF), and outperforms the autoregressive-model-based KFs future in the literature.

Many inference algorithms develop the time-domain connection of the direct by employ a Kalman filter base on a first-order (or sometimes second-order) estimate of the time-varying channel amid a norm based on link toning (CM), or on the Minimization of Asymptotic Variance (MAV). To decrease the complexity of the high-dimensional RW-KF for combined opinion of the multi-path multifaceted amplitudes, we suggest using an inferior dimensional RW-KF that estimate the compound amplitude of each path separately. We demonstrate that this amounts to a simplification of the joint multi-path Kalman increase formulation through the Woodbury’s identities. Hence, this innovative algorithm consists of a superposition of self-determining single-path single-carrier KFs, which be optimized in our earlier studies. This examination allow us to settle in the optimization to the authentic multi-path multi-carrier scenario, to afford logical formulas for the mean-square error presentation and the best tuning of the future estimator in a straight line as a function of the physical parameter of the canal (Doppler frequency, signal-to-noise-ratio, power delay profile). These logical formula are known for the first-, second-, and third-order RW models used in the KF. The future per-path KF is exposed to be as well-organized as the accurate KF (i.e., the joint multi-path KF), and outperforms the autoregressive-model-based KFs future in the literature.

Purpose The purpose of this study is to understand experimentally the mixing characteristics of a two-stream exhaust system with a supersonic Mach 1.5 primary jet that exits the rectangular C-D nozzle surrounded by a sonic secondary jet from a convergent rectangular nozzle by varying the aspect ratio (AR = 2 and 3) similar to those that can be available for future high-speed commercial aircraft. Design/methodology/approach This paper focuses on the experimental results of effects of AR at various expansion levels of jets issued/delivered from a central rectangular convergent-divergent nozzle of AR 2 and 3 surrounded by a coflow from a convergent rectangular sonic nozzle. The lip thickness of the primary nozzle is 2.2 mm. various nozzle pressure ratios (NPRs) ranging from 2, 3, 3.69 and 4 were chosen for pressure measurements. Findings For all the NPRs, AR 3 had a shorter core than AR 2. Also, AR 3 was found to decay faster in the transition and fully developed zones. The lateral plots show that the AR has an influence on the jet spread. Originality/value The structure of waves existing in the potential core of the rectangular coflow jet along with the major and minor axis planes was visualized by the shadowgraph technique.

An experimental investigation on the mixing characteristics of Mach 1.5 rectangular jet emanating from aspect ratios (ARs) 2 and 3 convergent-divergent nozzles is carried out at nozzle pressure ratios (NPRs) 3, 3.69, 4 and 5, respectively. At overexpanded (NPR3), correctly expanded (NPR 3.69) and underexpanded (NPRs 4, 5) levels of Mach 1.5 jet, the Pitot pressure measured along the centerline of AR2 and AR3 rectangular jets showed that the influence of aspect ratio on the jet mixing is not as strong as expected. The effect of aspect ratio is noticeable only in the core of both the jets. The core length of AR2 jet is shorter than AR3 jet at all the NPRs, which indicates faster mixing of AR2 jet with ambient fluid as compared to AR3 jet. But after core, the decay of AR2 and AR3 jets in the characteristic decay region remained almost same at all the NPRs except at correctly expanded NPR of 3.69, the AR3 jet decays slightly faster than AR2 jet. For all the cases, the jets have become fully developed together at far downstream location. The pressure profiles taken along major axis and minor axis of Mach 1.5 rectangular jets confirm the faster spreading of AR2 and AR3 jets along minor axis than along major axis. Both the jets are found to switch axes between $8D_{e}$ to $10D_{e}$ . The shadowgraph visualization reveals the complex structure of waves present in the major axis and minor axis planes of AR2 and AR3 rectangular jets.

This study investigates the effect of fin length on solidification characteristics of deionized water filled in a stainless steel spherical capsule immersed in a constant temperature bath. Alongside, Box-Benkehn experimental design is employed to develop mathematical models for predicting solidification duration using response surface regression. Experiments are carried out at three different bath temperatures (−6°C − 9°C and −12°C). Four radial fins of 3 mm diameter made of copper are fixed on the inner surface of the spherical capsule. Three fin-lengths of 7.5 mm, 13.5 mm, and 19.5 mm are chosen which corresponds to the annulus volume margins of 50%, 75%, and 90% respectively measured from the inner surface of the capsule. Results revealed that the fin length has no significant effect on the duration of solidification. The individual as well as interactive effects of frozen mass fraction and bath temperature were found to be statistically significant. The models developed were found to be statistically significant at a 99% confidence level and the confirmatory experiments validated that the developed models were able to predict the solidification time with reasonable accuracy. It was found that there is an improvement in solidification duration in the range between 17% and 21% using the combinations predicted by the desirability approach when compared to a capsule fitted with no fin under similar conditions. The spherical capsule fitted with a fin of length 16.5 mm shows a reduction of 21% in solidification duration to achieve 96% of frozen mass at a bath temperature of −12°C.

The need energy is more significant human lives.Different conventional and non-conventional energy resources provide the energy for world requirement.Butthe major problem associated with conventional energy resources are exhaustible and pollute the environment seriously. So researchers are focusing on nonconventional energy resources as more availability and environment friendly. Radiation from solar is freely available in major parts of the world in the day time.By usingsolarpanels, light energycan easilybe converted into electrical energyanddoany mechanicalwork by using that electrical energy. Agriculture plays a vital role in the Indian economy, new innovations gives full support to the farmers as well. In this article, a farmer friendly cost effective solar-operatedpesticidesprayer with additional applications like LED andmobilechargersare developed. From this environment-friendly proposed system, usage of fossil fuel is avoided with no operating cost and save the environment from pollution by notradiating the destructivesubstances like conventional systems. This proposed system can remotely access in various places like farm, garden, municipality, etc

The study is mainly to investigate the use of straight Simarouba oil as an alternate in the DI compression ignition engine and also the effect of hydrogen induction. Simarouba fuel is used as pilot fuel and hydrogen is used as secondary fuel which is inducted in inlet manifold by small modification in the CI engine. The mass share of hydrogen induction is fixed to maximum limit with the use of a manometer and for safety two flame arresters are used to avoid backfire in the hydrogen fuel line. The maximum hydrogen mass share percentage for diesel, B20, and B40 are 12.20%, 13.05% and 10.88% is fixed in full load condition. In maximum mass share, the BTE at full load conditions for diesel, B20, and B40 are 34.80%, 33.10%, and 29.65%. The obtained value of B20 at 13.05% mass share is nearly equal to the diesel value (0% mass share of hydrogen). The major drawback of increasing the mass share of hydrogen will increase nitric oxide emission due to the high heating value of hydrogen. The nitric oxide is increasing around 18% in the maximum mass share of hydrogen for B20 compare with diesel (0% mass share of hydrogen). But, there is a decreasing trend for other emissions like CO, HC, and smoke. This may be due to the absence of a carbon molecule in hydrogen fuel. The blended fuel B20 with maximum mass share obtain a better result than diesel with 0% mass share of hydrogen.

Arcs of conics are natural looking, smooth and gradual in curvature transition. The method in this paper proposes piecewise interpolating splines, which fit elliptic arcs between consecutive points in a given set of planar data points. Tangents of the curve at the data points are determined by a local procedure inspired by Akima's formulation. Elliptic arcs that suitably fit the orientation of these tangents are fit at all segments between control points, giving a G1-continuous curve fit. In another option, the curve segments on both sides of the control points are made curvature continuous by blending with circular arcs; resulting in a G2-continuous curve. These curves avoid unwanted inflections and undulations; and are capable of recognizing and generating straight-line paths and sharp corners; and also have an option to specify tension of curve at individual segments. Moreover, this is computationally less expensive. Having these desirable properties, this method can be used as an effective curve fitting tool for multiple-valued functional approximation, motion path tracing, 2D geometric modeling (including typography), etc.

Ancient DNA and RNA are valuable data sources for a wide range of disciplines. Within the field of ancient metagenomics, the number of published genetic datasets has risen dramatically in recent years, and tracking this data for reuse is particularly important for large-scale ecological and evolutionary studies of individual taxa and communities of both microbes and eukaryotes. AncientMetagenomeDir (archived at 10.5281/zenodo.3980833) is a collection of annotated metagenomic sample lists derived from published studies that provide basic, standardised metadata and accession numbers to allow rapid data retrieval from online repositories. These tables are community-curated and span multiple sub-disciplines to ensure adequate breadth and consensus in metadata definitions, as well as longevity of the database. Internal guidelines and automated checks facilitate compatibility with established sequence-read archives and term-ontologies, and ensure consistency and interoperability for future meta-analyses. This collection will also assist in standardising metadata reporting for future ancient metagenomic studies., Measurement(s) genome • Metagenome • Metadata • Ancient DNA Technology Type(s) digital curation Factor Type(s) geographic location • sample age Machine-accessible metadata file describing the reported data: 10.6084/m9.figshare.13241537

Herpes simplex virus type-2 (HSV-2) specific glycoprotein G (gG-2) is widely used as the antigen of choice for serodiagnosis of HSV-2. In order to develop an ELISA for serodetection of HSV-2 IgG in patient sera, the soluble form of the mature gG-2 antigen (mgG-2), gG283-649, was expressed using a baculovirus expression system. gG283-649 contains the complete extracellular domain of mgG-2 including the C-terminal region, which despite homology to gG-1, does not cross-react with HSV-1 antibodies present in HSV-1 positive patient sera. gG283-649 had increased performance compared to a previously described gG-2 fragment and showed high sensitivity and specificity in a method comparison with HerpeSelect 1 & 2 Immunoblot IgG, a commercially available FDA-cleared assay for serodetection of HSV-1 and 2 antibodies. A total of 234 clinical samples consisting of 134 high risk samples, including 45 samples from pregnant subjects, and a panel of 100 mixed diagnosis samples, spanning the measurable range were tested in the method comparison. Clinical sensitivity and specificity were determined to be 94.2% and 100%, respectively. We conclude that this soluble form of mgG-2 is a novel antigen of choice for developing an ELISA for type-specific serodiagnosis of HSV-2.

The ability to perform laboratory testing near the patient and with smaller blood volumes would benefit patients and physicians alike. We describe our design of a miniaturized clinical laboratory system with three components: a hardware platform (ie, the miniLab) that performs preanalytical and analytical processing steps using miniaturized sample manipulation and detection modules, an assay-configurable cartridge that provides consumable materials and assay reagents, and a server that communicates bidirectionally with the miniLab to manage assay-specific protocols and analyze, store, and report results (i.e., the virtual analyzer). The miniLab can detect analytes in blood using multiple methods, including molecular diagnostics, immunoassays, clinical chemistry, and hematology. Analytical performance results show that our qualitative Zika virus assay has a limit of detection of 55 genomic copies/ml. For our anti-herpes simplex virus type 2 immunoglobulin G, lipid panel, and lymphocyte subset panel assays, the miniLab has low imprecision, and method comparison results agree well with those from the United States Food and Drug Administration-cleared devices. With its small footprint and versatility, the miniLab has the potential to provide testing of a range of analytes in decentralized locations.

For smart structure and system, the material should be able to withstand higher load and the weight of the materials should be minimum. In the regard of less weight requirement aluminum and magnesium alloy and composites plays major role. This article gives understanding of preparation of magnesium composite for smart structures. Magnesium composite is a prospective resource for numerous solicitations of industries due to their good mechanical and physical characteristic. The upgrading of corrosion resistance, stiffness, specific strength, damping behavior, creep, fatigue and wear behavior characteristic are considerably induced by the addition of strengthening elements. This paper presents the synopsis on the effects of different casting techniques, their method, effect of methods, importance of their qualities and disadvantages. The main trend like the effects in mechanical properties, microstructure, agglomerating effect, interfacial transition, matrix -fiber bonding and the difficulties allied with the particle distribution by manufacturing process of smart material for smart structure are discussed in this paper

The ability to perform laboratory testing near the patient and with smaller blood volumes would benefit patients and physicians alike. We describe our design of a miniaturized clinical laboratory system with three components: a hardware platform (ie, the miniLab) that performs preanalytical and analytical processing steps using miniaturized sample manipulation and detection modules, an assay‐configurable cartridge that provides consumable materials and assay reagents, and a server that communicates bidirectionally with the miniLab to manage assay‐specific protocols and analyze, store, and report results (i.e., the virtual analyzer). The miniLab can detect analytes in blood using multiple methods, including molecular diagnostics, immunoassays, clinical chemistry, and hematology. Analytical performance results show that our qualitative Zika virus assay has a limit of detection of 55 genomic copies/ml. For our anti‐herpes simplex virus type 2 immunoglobulin G, lipid panel, and lymphocyte subset panel assays, the miniLab has low imprecision, and method comparison results agree well with those from the United States Food and Drug Administration‐cleared devices. With its small footprint and versatility, the miniLab has the potential to provide testing of a range of analytes in decentralized locations.

Digital marketplaces are disrupting the established traditional marketplace models and transforming the economy and societies in India, by providing a platform for innovations. This disruption impacts drastically the way Indians’ shop, bank, work, book their holidays, and even hailing a cab. India’s unique demographic dividend with 50% of the 1.2 billion population less than 25 years, rising middle class income, and the mobile and internet growth are fueling the highest e‐commerce growth rate in the world. This paper analyses the revolutionary growth of digital marketplace in India based on the four key pillars of digital marketplace, namely payment landscape, logistics, mobile/internet infrastructure and business and revenue model, and illustrates how these pillars are undergoing innovations and transformations. The paper also analyzes the key challenges to digital marketplace growth and illustrates how various organizations have made use of the unique opportunities provided by India, with appropriate examples. Entry of major players such as Alibaba and Rakuten are expected to make the online marketplace more competitive and propel India to enter into the third wave of Digital Marketplace 3.0 – the golden era of Digital Marketplace in India. The innovative practices, processes and models adopted by the various firms analyzed in this paper could serve as a template for digital marketplace evolution in emerging markets.

Enamel thickness varies substantially among extant hominoids and is a key trait with significance for interpreting dietary adaptation, life history trajectory, and phylogenetic relationships. There is a strong link in humans between enamel formation and mutations in the exons of the four genes that code for the enamel matrix proteins and the associated protease. The evolution of thick enamel in humans may have included changes in the regulation of these genes during tooth development. The cis-regulatory region in the 5' flank (upstream non-coding region) of MMP20, which codes for enamelysin, the predominant protease active during enamel secretion, has previously been shown to be under strong positive selection in the lineages leading to both humans and chimpanzees. Here we examine evidence for positive selection in the 5' flank and 3' flank of AMELX, AMBN, ENAM, and MMP20. We contrast the human sequence changes with other hominoids (chimpanzees, gorillas, orangutans, gibbons) and rhesus macaques (outgroup), a sample comprising a range of enamel thickness. We find no evidence for positive selection in the protein-coding regions of any of these genes. In contrast, we find strong evidence for positive selection in the 5' flank region of MMP20 and ENAM along the lineage leading to humans, and in both the 5' flank and 3' flank regions of MMP20 along the lineage leading to chimpanzees. We also identify putative transcription factor binding sites overlapping some of the species-specific nucleotide sites and we refine which sections of the up- and downstream putative regulatory regions are most likely to harbor important changes. These non-coding changes and their potential for differential regulation by transcription factors known to regulate tooth development may offer insight into the mechanisms that allow for rapid evolutionary changes in enamel thickness across closely-related species, and contribute to our understanding of the enamel phenotype in hominoids.

The EF-hand protein, DREAM/KChIP3 (henceforth referred to as DREAM), regulates apoptosis by incompletely understood mechanisms. We demonstrate that in the presence of Ca2+, DREAM interacts with hexokinase I, a protein known to bind mitochondria and regulate apoptosis. A mutant DREAM protein construct incapable of binding Ca2+ does not associate with hexokinase I. The amino-terminal portion of DREAM is required for binding to hexokinase I, as a DREAM construct lacking the first 94 amino terminal residues fails to bind hexokinase I. Expression of DREAM in neuroblastoma cells enhances cisplatin mediated caspase-3 activity. Simultaneous expression of hexokinase I in such cells reduces DREAM-stimulated apoptosis. DREAM overexpression in neuroblastoma cells reduces hexokinase I localization on isolated mitochondria. The interaction of DREAM with hexokinase I may be important in the regulation of neuronal apoptosis.

As methodologies and tools for chip-level design mature, design effort becomes focused on increasingly higher levels of abstraction. We present a tutorial on a design methodology for chip and system design and present a test case that justifies the future goal of a 100 h design cycle.

Downstream regulatory element antagonistic modulator (DREAM/KChIP3), a neuronal EF-hand protein, modulates pain, potassium channel activity, and binds presenilin 1. Using affinity capture of neuronal proteins by immobilized DREAM/KChIP3 in the presence and absence of calcium (Ca(2+)) followed by mass spectroscopic identification of interacting proteins, we demonstrate that in the presence of Ca(2+), DREAM/KChIP3 interacts with the EF-hand protein, calmodulin (CaM). The interaction of DREAM/KChIP3 with CaM does not occur in the absence of Ca(2+). In the absence of Ca(2+), DREAM/KChIP3 binds the EF-hand protein, calcineurin subunit-B. Ca(2+)-bound DREAM/KChIP3 binds CaM with a dissociation constant of ∼3 μM as assessed by changes in DREAM/KChIP3 intrinsic protein fluorescence in the presence of CaM. Two-dimensional (1)H,(15)N heteronuclear single quantum coherence spectra reveal changes in chemical shifts and line broadening upon the addition of CaM to (15)N DREAM/KChIP3. The amino-terminal portion of DREAM/KChIP3 is required for its binding to CaM because a construct of DREAM/KChIP3 lacking the first 94 amino-terminal residues fails to bind CaM as assessed by fluorescence spectroscopy. The addition of Ca(2+)-bound DREAM/KChIP3 increases the activation of calcineurin (CN) by calcium CaM. A DREAM/KChIP3 mutant incapable of binding Ca(2+) also stimulates calmodulin-dependent CN activity. The shortened form of DREAM/KChIP3 lacking the NH(2)-terminal amino acids fails to activate CN in the presence of calcium CaM. Our data demonstrate the interaction of DREAM/KChIP3 with the important EF-hand protein, CaM, and show that the interaction alters CN activity.

Differential hydrogen/deuterium exchange (H/DX) coupled with mass spectrometry (H/DX-MS) offers a rapid and sensitive characterization of changes in proteins following perturbations induced by changes in folding, ligand binding, oligomerization, and modification. The characterization of H/DX rates by software tools and automated data processing often relies on the centroid mass calculation and, thereby, the deuterium distribution in the mass spectra is neglected. Here we present an example demonstrating the clear limitation of using only a centroid approach to characterize the H/DX rate, in which the change in protein is not reflected as the difference in deuterium uptake based on centroid calculation.

The signaling state of the photoactive yellow protein (PYP) photoreceptor is transiently developed via isomerization of its blue-light-absorbing chromophore. The associated structural rearrangements have large amplitude but, due to its transient nature and chemical exchange reactions that complicate NMR detection, its accurate three-dimensional structure in solution has been elusive. Here we report on direct structural observation of the transient signaling state by combining double electron electron resonance spectroscopy (DEER), NMR, and time-resolved pump probe X-ray solution scattering (TR-SAXS/WAXS). Measurement of distance distributions for doubly spin-labeled photoreceptor constructs using DEER spectroscopy suggests that the signaling state is well ordered and shows that interspin-label distances change reversibly up to 19 angstrom upon illumination. The SAXS/WAXS difference signal for the signaling state relative to the ground state indicates the transient formation of an ordered and rearranged conformation, which has an increased radius of gyration, an increased maximum dimension, and a reduced excluded volume. Dynamical annealing calculations using the DEER derived long-range distance restraints in combination with short-range distance information from H-1-N-15 HSQC perturbation spectroscopy give strong indication for a rearrangement that places part of the N-terminal domain in contact with the exposed chromophore binding cleft while the terminal residues extend away from the core. Time-resolved global structural information from pump probe TR-SAXS/WAXS data supports this conformation and allows subsequent structural refinement that includes the combined energy terms from DEER, NMR, and SAXS/WAXS together. The resulting ensemble simultaneously satisfies all restraints, and the inclusion of TR-SAXS/WAXS effectively reduces the uncertainty arising from the possible spin-label orientations. The observations are essentially compatible with reduced folding of the I-2' state (also referred to as the 'pB' state) that is widely reported, but indicates it to be relatively ordered and rearranged. Furthermore, there is direct evidence for the repositioning of the N-terminal region in the I-2' state, which is structurally modeled by dynamical annealing and refinement calculations.

The main focus of leprosy control has been case detection and treatment delivery with relative neglect of prevention of disability. Absence of reliable data and lack of research have added to the problem. This raised concerns about the capacity of the general health system to address the needs of people living with leprosy-related disabilities. In this prospective study appropriate services for people living with leprosy-related disabilities were introduced in the form of self-care training, guidance and monitoring by the general health staff facilitated by a non-governmental organisation leprosy centre in a district in south India with a population of 3.1 million (estimated in 2005). The staff identified 1232 people with leprosy-related disabilities and trained them in self-care. Follow-up assessments indicated that 86% were found to be practising self-care regularly and all the 239 general health workers were found to be actively involved. The most heartening outcome was the healing of plantar ulcers in 70% of people at the 1-year follow up. This intervention is sustainable because of the simplicity of the procedures and the involvement of all health staff including supervisors.

A novel class of synthetic peptides has been designed that inhibit the thrombin catalytic site and exhibit specificity for the anion-binding exosite (ABE) of {alpha}-thrombin. These peptides, called hirulogs, consist of (i) an active-site specificity sequence with a restricted Arg-Pro scissile bond, (ii) a polymeric linker of glycyl residues from 6 to 18 {angstrom} in length, and (iii) an ABE recognition sequence such as that in the hirudin C-terminus. Hirulog-1 ((D-Phe)-Pro-Arg-Pro-(Gly){sub 4}-Asn-Gly-Asp-Phe-Glu-Glu-Ile-Pro-Glu-Tyr-Leu) inhibits the thrombin-catalyzed hydrolysis of a tripeptide p-nitroanilide substrate with K{sub i} = 2.3 nM. In contrast, the synthetic C-terminal hirudin peptide S-Hir{sub 53-64}, which binds to the thrombin ABE, blocked the fibrinogen clotting activity of the enzyme with K{sub i} = 144 nM but failed to inhibit the hydrolysis of p-nitroanilide substrates at concentrations as high as 1 mM. Hirulog-1, but not S-Hir{sub 53-64}, was found to inhibit the incorporation of ({sup 14}C)diisopropyl fluorophosphate in thrombin. Hirulog-1 appears specific for thrombin as it lacks inhibitory activities toward human factor Xa, human plasmin, and bovine trypsin at inhibitor:enzyme concentrations 3 orders of magnitude higher than those required to inhibit thrombin. The optimal inhibitory activity of hirulog-1 depends upon all three components of its structure. Comparison of anticoagulant activitiesmore » of hirulog-1, hirudin, and S-Hir{sub 53-64} showed that the synthetic hirulog-1 is 2-fold more potent than hirudin and 100-fold more active than S-Hir{sub 53-64} in increasing the activated partial thromboplastin time of normal human plasma.« less

We have coupled a chemiluminescent detection method that uses an alkaline phosphatase label to the genomic DNA sequencing protocol of Church and Gilbert [Church, G. M. & Gilbert, W. (1984) Proc. Natl. Acad. Sci. USA 81, 1991-1995]. Images of sequence ladders are obtained on x-ray film with exposure times of less than 30 min, as compared to 40 h required for a similar exposure with a 32P-labeled oligomer. Chemically cleaved DNA from a sequencing gel is transferred to a nylon membrane, and specific sequence ladders are selected by hybridization to DNA oligonucleotides labeled with alkaline phosphatase or with biotin, leading directly or indirectly to deposition of enzyme. If a biotinylated probe is used, an incubation with avidin-alkaline phosphatase conjugate follows. The membrane is soaked in the chemiluminescent substrate (AMPPD) and is exposed to film. Dephosphorylation of AMPPD leads in a two-step pathway to a highly localized emission of visible light. The demonstrated shorter exposure times may improve the efficiency of a serial reprobing strategy such as the multiplex sequencing approach of Church and Kieffer-Higgins [Church, G. M. & Kieffer-Higgins, S. (1988) Science 240, 185-188].

Summary form only given. Recent advances in semiconductor technology have made it possible to integrate multi million transistors on a single chip; design and verification teams face several challenges managing the complexity. Some of these challenges include, specification and verification at the functional level, closing early on the system level architecture, extensive simulations of the hardware and software components and finalizing the path to implementation of the entire SOC. This tutorial covers the state-of-the-art in specification, design and verification techniques.

The authors describe a novel algorithm that combines the hardware scheduling and component selection phases for high level synthesis. The algorithm improves on previous work in scheduling, by being able to simultaneously select components from a given library. This enlarges the design space, resulting in better optimized designs. Experimental results on the elliptic filter benchmark demonstrate that exploiting all available components in the library results in designs with smaller area compared to designs produced by scheduling with a single implementation for each component type. >

To determine the type and incidence of adverse events seen in the first 13 months of extended wear from a prospective clinical trial involving the use of disposable hydrogels on a 6-night extended wear and replacement schedule.A prospective, noncomparative clinical trial.Three hundred thirty subjects were enrolled in the study from March 1993 to August 1996 and commenced in extended wear. The subjects were new contact lens users with ages ranging from 16 to 39 years and refractive errors from -1.00 to -6.50-diopter (D) sphere with cylindrical component less than 1.0 D.Disposable hydrogel lenses on a 6-night extended wear and replacement schedule.To determine the incidence of adverse events in the first 13 months of lens wear with the use of disposable hydrogels on an extended wear schedule. In addition, corneal infiltrative events observed with lens wear were classified based on their presenting signs and symptoms.A total of 137 adverse events were seen in 102 subjects in the first 13 months of extended wear. The type of adverse events and the incidence per 100 patients per year of lens wear were as follows: corneal infiltrative events (44.4 events), contact lens papillary conjunctivitis (6.4 events), neovascularization and corneal wrinkling (1.7 events each), corneal striae and superior epithelial arcuate lesions (1.3 events each), and blepharokeratoconjunctivitis (0.4 event). Viral keratoconjunctivitis was also observed (0.8 event). The corneal infiltrative events were further subclassified as follows: contact lens-induced peripheral ulcers (13.6 events), contact lens-induced acute red eyes (12.3 events), infiltrative keratitis (9.7 events), and asymptomatic infiltrative events (8.9 events). There were no events of infectious keratitis. Except for all events of contact lens-induced peripheral ulcers and two isolated events of infiltrative keratitis that resulted in scarring, all of the corneal infiltrative events resolved without sequelae. None of the events caused any loss of best-corrected visual acuity.Based on the observations from the first 13 months of extended wear in the trial, the majority of the complications associated with extended wear of disposable hydrogels are corneal infiltrative events. No events of microbial keratitis were seen in the first 13 months of extended wear. None of the events were associated with loss in best-corrected visual acuity. Clearly, frequent and regular disposing of lenses does not eliminate adverse effects, and better materials and designs are required for extended wear to be a successful method.

This study was undertaken to determine the extent of adhesion of cysts and trophozoites of Acanthamoeba to worn disposable hydrogel lenses and to study the effect of time, lens material, and washing on the adhesion.Etafilcon A (58% water content) and Polymacon (38% water content) lenses worn on a 6 night extended wear basis and collected aseptically in PBS pH 7.2, were used for the study. Unworn lenses of the same materials were used as controls. Segments of lenses were incubated in Acanthamoeba suspension (cysts and trophozoites) and examined at 0, 5, and 24 hours, either washed or unwashed, for adherent Acanthamoeba. The amoebae adherent per square mm surface of the lens were enumerated. Levels of adhesion in various groups were compared using Wilcoxon sign rank test.At 0 hours, unwashed worn Polymacon lenses showed significantly greater adherence than Etafilcon A lenses compared to the controls (P = 0.0469). However, there was no difference at 5 and 24 hours exposure time between the lenses. At 24 hours the adhesion of Acanthamoeba was greater for washed worn lenses of both types compared to washed lenses of controls (P = 0.002, 0.012).Amoebae adhered with equal affinity to both types of lenses but adhered with greater affinity to worn lenses than unworn lenses. Adhesion of Acanthamoeba to worn disposable lenses may be a factor in the pathogenesis of Acanthamoeba keratitis in disposable lens wearers.

There are couples with unmet family planning needs and couples who do not use any modern method, yet they desire to space or avoid pregnancies. Many of them look for safe and effective options like the natural family planning methods. The Billings Ovulation Method based on single index cervical mucus parameter is one such option. The present multicentre trial conducted in India has shown an encouraging use-effectiveness of the method, indicating method failure as low as 1.5 +/- 0.3 and use-failure 15.9 +/- 0.8 per 100 users at 21 months. The method continuation rates have also been as high as 88.3/100 users at 6 months and 52.0/100 users at 21 months.

Contact lens related Acanthamoeba keratitis has been more often associated with the use of soft contact lenses than with the use of rigid gas permeable (RGP) contact lenses. We studied the adherence of Acanthamoeba cysts and trophozoites to unworn RGP (silicone acrylate and fluoropolymer) and low and high water content soft contact lenses marketed in India. A human corneal isolate of Acanthamoeba polyphaga suspended in saline (1 x 10(5) amoebae/mL) was used. Lens segments were exposed to A. polyphaga cysts or trophozoites with or without shaking. After exposure for zero to 7 hours, lenses were either washed or not washed and the adherence was determined using light microscopy and a hemocytometer (per square millimeter of the lens). Both cysts and trophozoites immediately adhered to all four types of lenses. Adhesion increased with an increase in exposure time. The number of adherent cysts and trophozoites was higher (P = 0.043-0.0003) in the unwashed group of lenses at all exposure times for all lenses. Among washed groups, adherence was greatest with RGP silicone acrylate and high water content soft lenses. Trophozoites of Acanthamoeba showed greater adherence to RGP lenses, whereas cysts showed greater adherence to high water content soft lenses. Shaking did not have a significant effect on adherence. Adhesion of Acanthamoeba to contact lenses may play an important role in the pathogenesis of Acanthamoeba keratitis.

Describes a system called CLSS, that serves as a workbench for the synthesis of control logic. The system provides an integrated solution to the designer by providing effective solutions for all the major steps in the control logic synthesis flow. CLSS allows specification of the control logic in the form of a state transition representation. Through a set of transformation steps, all of which are an integral part of the CLSS workbench, the layout of the design is generated for standard cell or PLA based implementations. In addition to providing automated solutions to optimally perform state assignment, logic minimization, constrained area-delay optimization, etc., CLSS provides a mechanism for the expert designers to override the decisions taken by the system. The major strength of the system lies in the tight coupling between the several components of the flow in an integrated environment. Designed at Texas Instruments, the CLSS workbench has been successfully used on several large and realistic designs. >

Background: An abnormal karyotype in either partner, especially featuring a translocation and/or inversion is considered to be a cause of recurrent miscarriages. It is generally assumed that recurrent miscarriage might be due to recurrent chromosomal abnormalities in the fetus due to a balanced aberration in one of the parents being inherited by the offspring in an unbalanced form. Aim: Evaluation of chromosomal rearrangements in couples with recurrent miscarriages. Materials and Methods: Peripheral blood was collected and lymphocyte cultures were set up. Slides prepared from the cell suspension were stained and screened for metaphases followed by karyotyping. Result: Balanced translocation was observed in the male partner in one case and in the female partners in the three other cases. Conclusion: Couples with recurrent miscarriage should be investigated for chromosomal rearrangements, thus helping in genetic counseling and providing the options for future pregnancies.

The genes that encode the α, β, and γ subunits of component C of m ethyl c oenzyme M r eductase ( mcrA, mcrB , and mcrG ) in Methanococcus vannielii have been cloned and sequenced, and their expression in Escherichia coli has been demonstrated. These genes are organized into a five-gene cluster, mcrBDCGA , which contains two genes, designated mcrC and mcrD , with unknown functions. The mcr genes are separated by very short intergenic regions that contain multiple translation stop codons and strong ribosomebinding sequences. Although the genome of M. vannielii is 69 mol% A+T, there is a very strong preference in the mcrA, mcrB , and mcrG genes for the codon with a C in the wobble position in the codon pairs AA U C (asparagine), GA U C (aspartic acid), CA U C (histidine), AU U C (isoleucine), UU U C (phenylalanine), and UA U C (tyrosine). The mcrC and mcrD genes do not show this codon preference and frequently have U or A in the wobble position. As the codon pairs listed above are likely to be translated by the same tRNA with a G in the first anticodon position, the presence of C in the wobble position might ensure maximum efficiency of translation of transcripts of these very highly expressed genes.

In the DNAs of all Ph1-positive chronic myelocytic leukemia patients studied to date, a breakpoint on chromosome 22 (the Ph1 chromosome) can be demonstrated with a probe from the bcr (breakpoint cluster region). Although the K562 cell line was established from cells of a chronic myelocytic leukemia patient, we have been unable to detect the Ph1 chromosome by cytogenetic means. Employing a probe from the 5' region of bcr, we have cloned an amplified Ph1 breakpoint fragment from K562. This demonstrates that K562 contains multiple remnants of a Ph1 chromosome with a breakpoint within bcr and thus may serve as a model system for the study of Ph1-positive chronic myelocytic leukemia at a molecular level. The isolation of bcr cDNA sequences shows that parts of bcr encode a protein. Employing K562, we demonstrate the presence of an abnormally sized mRNA species hybridizing to c-abl and to a bcr cDNA probe, indicating the possible consequence of the Ph1 translocation on a transcriptional level in chronic myelocytic leukemia. The isolation and sequencing of a cDNA containing the breakpoint area of this mRNA provide further evidence for its chimeric structure. Cloning of large stretches of chromosomal DNA flanking bcr and c-abl sequences in K562 and identification of the exons participating in the formation of the chimeric mRNA shows that a splice of at least 99 kilobases is made to fuse the 3' bcr exon to the 5' c-abl exon. Furthermore two chimeric cDNAs were isolated containing chromosome 9 sequences that map 43.5 kilobases downstream from the K562 breakpoint. These chromosome 9 sequences neither hybridize to the 8.5-kilobase chimeric c-abl mRNA nor to normal c-abl mRNAs in Hela cells and probably represent incorrect splicing products present in the K562 cell line.

Synthetic peptides based on the COOH-terminal 21 residues of hirudin were prepared in order to 1) evaluate the role of this segment in hirudin action toward thrombin, 2) define the shortest peptide derivative with anticoagulant activity, and 3) investigate the role of tyrosine sulfation in the peptides' inhibitory activities. A hirudin derivative of 20 amino acids, Hir45-64 (derived from residues 45-64 of the hirudin polypeptide), was found to effect a dose-dependent increase in the activated partial thromboplastin time (APTT) of normal human plasma but to have no measurable inhibitory activity toward thrombin cleavage of a tripeptidyl p-nitroanilide substrate. Anticoagulant activity in hirudin derivatives was comparable in peptides of 20, 16, and 12 residues truncated from the NH2 terminus. Additional truncated peptides prepared by synthesis and carboxypeptidase treatment reveal that the minimal sequence of a hirudin peptide fragment with maximal anticoagulant activity is contained within the sequence: NH2-Asn-Gly-Asp-Phe-Glu-Glu-Ile-Pro-Glu-Glu-Tyr-Leu-COOH. The 12-residue derivative thus identified was reacted with dicyclohexylcarbodiimide in the presence of sulfuric acid to yield a Tyr-sulfated peptide, S-Hir53-64. By comparison to unsulfated peptide, S-Hir53-64 was found to contain a specific inhibitory activity enhanced by one order of magnitude toward increase in APTT and to effect a dose-dependent increase in thrombin time of normal human plasma to yield a 4-fold increase in thrombin time with 2.5 micrograms/ml peptide using 0.8 units/ml alpha-thrombin. Comparison of S-Hir53-64 to hirudin in thrombin time and APTT assays reveals a 50-fold difference in molar specific activities toward inhibition of thrombin. Comparison of antithrombin activities of S-Hir53-64 using a variety of animal thrombins demonstrates greatest inhibitory activity toward murine, rat, and human enzymes and a 10-fold reduced activity toward bovine thrombin.

The previously assigned structure of human big gastrin is revised as a result of sequencing and immunological studies on synthetic peptides. A nonadecapeptide has been synthesized and found to have full immunochemical potency compared with natural human G34 in a radioimmunoassay which is specific for the N-terminal sequence. Syntheses of the peptides were achieved using the stepwise procedure with benzyloxycarbonyl-amino acids and fragment couplings mediated mainly by the dicyclohexylcarbodiimide procedure in the presence of either N-hydroxysuccinimide or 1-hydroxybenzotriazole. Purification of the peptide fragments was by Sephadex LH-20 chromatography and removal of protecting groups was effected using 90% trifluoroacetic acid in the presence of scavengers. Purification of the nonadecapeptide was achieved by high performance liquid chromatography.

This 4-year prospective randomized study is designed to compare the effectiveness and complication rates of the Nova T and MLCu250 inserted immediately postabortion. At the cut-off date (30 November 1983), all patients in the first cohort of 400 women had completed at least 12 months of use. At this stage of the trial, no significant differences had emerged between the two devices in any of the standard termination categories.

We have purified two 35 kd phospholipase A2 inhibitors from human placenta, which we refer to as lipocortin I and II. Both proteins exhibit similar biochemical properties and occur in placenta at about 0.2% of the total protein. By peptide mapping, sequence, and immunological analyses, we show that lipocortin I and the 35 kd substrate for the EGF-receptor/kinase from A431 cells are the same protein. By similar criteria, we determine that lipocortin II is the human analogue of pp36, a major substrate for pp60src, which has been characterized in chicken embryo fibroblasts and in bovine brush border preparations. The amino acid sequences of lipocortin I and II that we deduced from cDNA clones share 50% homology, indicating that they probably evolved from a common gene.

We have deduced the 1675-amino acid sequence of rat clathrin heavy chain from cDNA clones and predict a protein of Mr 191,569. We have established the polarity of the heavy chain and assigned sequence positions to several structural landmarks of the clathrin leg. The terminal domain at the distal end of the clathrin leg is at the amino terminus of the heavy chain. It is connected to the distal segment by a flexible "link" from Tyr-479 to Arg-523. There is an unusual sequence at the carboxyl terminus that may form the globular projection at the vertex of the clathrin trimer. We suggest that a possible site of heavy-chain-light-chain interaction is located in the proximal segment. Comparison with other partially sequenced mammalian clathrin heavy chains shows that the primary structure is highly conserved. The heavy chain is unrelated to other classes of structural proteins.

Molecular analysis of the heterodimeric T-cell antigen receptor of insulin-specific class II-restricted T-cell hybridomas (THys) derived from C57BL/6 (B6) wild-type and B6.C-H-2bm12 (bm12) mutant mice revealed that such T cells use a diverse V gene repertoire. Analysis of three THys that use related V genes, however, showed a number of novel features. Two THys that share major histocompatibility complex restriction use V alpha genes that are 98.6% homologous. Two THys sharing the same antigen fine specificity use a particular germ line V beta D beta J beta combination. A 21-base-pair deletion in the 5' segment of the J beta gene occurs in one THy, suggesting a novel mechanism for generating diversity in T-cell antigen receptor beta genes. The first amino acid encoded by N sequences at the V-D junction is conserved in a pair of T cells which recognize identical antigenic epitopes. The implications of these findings for the structural mechanisms underlying major histocompatibility complex-restricted antigen-specific T-cell recognition are discussed.

Molecular analysis of the heterodimeric T-cell antigen receptor of insulin-specific class II-restricted T-cell hybridomas (THys) derived from C57BL/6 (B6) wild-type and B6.C-H-2bm12 (bm12) mutant mice revealed that such T cells use a diverse V gene repertoire. Analysis of three THys that use related V genes, however, showed a number of novel features. Two THys that share major histocompatibility complex restriction use V alpha genes that are 98.6% homologous. Two THys sharing the same antigen fine specificity use a particular germ line V beta D beta J beta combination. A 21-base-pair deletion in the 5' segment of the J beta gene occurs in one THy, suggesting a novel mechanism for generating diversity in T-cell antigen receptor beta genes. The first amino acid encoded by N sequences at the V-D junction is conserved in a pair of T cells which recognize identical antigenic epitopes. The implications of these findings for the structural mechanisms underlying major histocompatibility complex-restricted antigen-specific T-cell recognition are discussed.

The clathrin light chains fall into two major classes, LCA and LCB. In an intact clathrin triskelion, one light chain, of either class, is bound to the proximal segment of a heavy chain leg. Analysis of rat brain and liver complementary DNA clones for LCA and LCB shows that the two light chain classes are closely related. There appear to be several members of each class having deletions of varying length aligned at the same position. A set of ten heptad elements, characteristic of alpha-helical coiled coils, is a striking feature of the central part of each derived amino acid sequence. These observations suggest a model in which the alpha-helical segment mediates binding to clathrin heavy chains and the amino- and carboxyl-terminal segments mediate interactions with other proteins. They also suggest an explanation for the observed tissue-dependent size variation for members of each class.