Proteinase inhibitor-inducing activity of the prohormone prosystemin resides exclusively in the C-terminal systemin domain

Prosystemin is the 200-amino acid precursor of the 18-amino acid polypeptide defense hormone, systemin. Herein, we report that prosystemin was found to be as biologically active as systemin when assayed for proteinase inhibitor induction in young tomato plants and nearly as active in the alkalinization response in Lycopersicon esculentum suspension-cultured cells. Similar to many animal prohormones that harbor multiple signals, the systemin precursor contains five imperfect repetitive domains N-terminal to a single systemin domain. Whether the five repetitive domains contain defense signals has not been established. N-terminal deletions of prosystemin had little effect on its activity in tomato plants or suspension-cultured cells. Deletion of the C-terminal region of prosystemin containing the 18-amino acid systemin domain completely abolished its proteinase inhibitor induction and alkalinization activities. The apoplastic fluid from tomato leaves and the medium of cultured cells were analyzed for proteolytic activity that could process prosystemin to systemin. These experiments showed that proteolytic enzymes present in the apoplasm and medium could cleave prosystemin into large fragments, but the enzymes did not produce detectable levels of systemin. Additionally, inhibitors of these proteolytic enzymes did not affect the biological activity of prosystemin. The cumulative data indicated that prosystemin and/or large fragments of prosystemin can be active inducers of defense responses in both tomato leaves and suspension-cultured cells and that the only region of prosystemin that is responsible for activating the defense response resides in the systemin domain.

Neuronal type information encoded in the basic-helix–loop–helix domain of proneural genes

The proneural genes encode basic-helix–loop–helix (bHLH) proteins and promote the formation of distinct types of sensory organs. In Drosophila, two sets of proneural genes, atonal (ato) and members of the achaete–scute complex (ASC), are required for the formation of chordotonal (ch) organs and external sensory (es) organs, respectively. We assayed the production of sensory organs in transgenic flies expressing chimeric genes of ato and scute (sc), a member of ASC, and found that the information that specifies ch organs resides in the bHLH domain of ato; chimeras containing the b domain of ato and the HLH domain of sc also induced ch organ formation, but to a lesser extent than those containing the bHLH domain of ato. The b domains of ato and sc differ in seven residues. Mutations of these seven residues in the b domain of ato suggest that most or perhaps all of these residues are required for induction of ch organs. None of these seven residues is predicted to contact DNA directly by computer simulation using the structure of the myogenic factor MyoD as a model, implying that interaction of ato with other cofactors is likely to be involved in neuronal type specification.

A Role for the GSG Domain in Localizing Sam68 to Novel Nuclear Structures in Cancer Cell Lines

The GSG (GRP33, Sam68, GLD-1) domain is a protein module found in an expanding family of RNA-binding proteins. The numerous missense mutations identified genetically in the GSG domain support its physiological role. Although the exact function of the GSG domain is not known, it has been shown to be required for RNA binding and oligomerization. Here it is shown that the Sam68 GSG domain plays a role in protein localization. We show that Sam68 concentrates into novel nuclear structures that are predominantly found in transformed cells. These Sam68 nuclear bodies (SNBs) are distinct from coiled bodies, gems, and promyelocytic nuclear bodies. Electron microscopic studies show that SNBs are distinct structures that are enriched in phosphorus and nitrogen, indicating the presence of nucleic acids. A GFP-Sam68 fusion protein had a similar localization as endogenous Sam68 in HeLa cells, diffusely nuclear with two to five SNBs. Two other GSG proteins, the Sam68-like mammalian proteins SLM-1 and SLM-2, colocalized with endogenous Sam68 in SNBs. Different GSG domain missense mutations were investigated for Sam68 protein localization. Six separate classes of cellular patterns were obtained, including exclusive SNB localization and association with microtubules. These findings demonstrate that the GSG domain is involved in protein localization and define a new compartment for Sam68, SLM-1, and SLM-2 in cancer cell lines.

Solution 19F nuclear Overhauser effects in structural studies of the cytoplasmic domain of mammalian rhodopsin

19F nuclear Overhauser effects (NOEs) between fluorine labels on the cytoplasmic domain of rhodopsin solubilized in detergent micelles are reported. Previously, high-resolution solution 19F NMR spectra of fluorine-labeled rhodopsin in detergent micelles were described, demonstrating the applicability of this technique to studies of tertiary structure in the cytoplasmic domain. To quantitate tertiary contacts we have applied a transient one-dimensional difference NOE solution 19F NMR experiment to this system, permitting assessment of proximities between fluorine labels specifically incorporated into different regions of the cytoplasmic face. Three dicysteine substitution mutants (Cys-140–Cys-316, Cys-65–Cys-316, and Cys-139–Cys-251) were labeled by attachment of the trifluoroethylthio group through a disulfide linkage. Each mutant rhodopsin was prepared (8–10 mg) in dodecylmaltoside and analyzed at 20°C by solution 19F NMR. Distinct chemical shifts were observed for all of the rhodopsin 19F labels in the dark. An up-field shift of the Cys-316 resonance in the Cys-65–Cys-316 mutant suggests a close proximity between the two residues. When analyzed for 19F-19F NOEs, a moderate negative enhancement was observed for the Cys-65–Cys-316 pair and a strong negative enhancement was observed for the Cys-139–Cys-251 pair, indicating proximity between these sites. No NOE enhancement was observed for the Cys-140–Cys-316 pair. These NOE effects demonstrate a solution 19F NMR method for analysis of tertiary contacts in high molecular weight proteins, including membrane proteins.

Role of the C2A domain of synaptotagmin in transmitter release as determined by specific antibody injection into the squid giant synapse preterminal.

Squid synaptotagmin (Syt) cDNA, including its open reading frame, was cloned and polyclonal antibodies were obtained in rabbits immunized with glutathione S-transferase (GST)-Syt-C2A. Binding assays indicated that the antibody, anti-Syt-C2A, recognized squid Syt and inhibited the Ca(2+)-dependent phospholipid binding to the C2A domain. This antibody, when injected into the preterminal at the squid giant synapse, blocked transmitter release in a manner similar to that previously reported for the presynaptic injection of members of the inositol high-polyphosphate series. The block was not accompanied by any change in the presynaptic action potential or the amplitude or voltage dependence of the presynaptic Ca2+ current. The postsynaptic potential was rather insensitive to repetitive presynaptic stimulation, indicating a direct effect of the antibody on the transmitter release system. Following block of transmitter release, confocal microscopical analysis of the preterminal junction injected with rhodamine-conjugated anti-Syt-C2A demonstrated fluorescent spots at the inner surface of the presynaptic plasmalemma next to the active zones. Structural analysis of the same preparations demonstrated an accumulation of synaptic vesicles corresponding in size and distribution to the fluorescent spots demonstrated confocally. Together with the finding that such antibody prevents Ca2+ binding to a specific receptor in the C2A domain, these results indicate that Ca2+ triggers transmitter release by activating the C2A domain of Syt. We conclude that the C2A domain is directly related to the fusion of synaptic vesicles that results in transmitter release.

A planar UWB antenna with signal rejection capability in the 4-6 GHz band

The design of a compact planar antenna featuring ultra wideband performance and simultaneous signal rejection in the 4-6 GHz band, assigned for IEEE802.11a and HIPERLAN/2, is presented. The design is demonstrated assuming RT6010LM substrate with a relative dielectric constant of 10.2 and thickness of 0.64 mm. The presented results show that the designed antenna of 27 mm * 20 mm dimensions has a bandwidth from 2.7 GHz to more than 10 GHz excluding the rejection band. The antenna features near omnidirectional characteristics and good radiation efficiency.

Grayscale and colour image Codec based on matching pursuit in the spatio-frequency domain

This report presents and evaluates a novel idea for scalable lossy colour image coding with Matching Pursuit (MP) performed in a transform domain. The benefits of the idea of MP performed in the transform domain are analysed in detail. The main contribution of this work is extending MP with wavelets to colour coding and proposing a coding method. We exploit correlations between image subbands after wavelet transformation in RGB colour space. Then, a new and simple quantisation and coding scheme of colour MP decomposition based on Run Length Encoding (RLE), inspired by the idea of coding indexes in relational databases, is applied. As a final coding step arithmetic coding is used assuming uniform distributions of MP atom parameters. The target application is compression at low and medium bit-rates. Coding performance is compared to JPEG 2000 showing the potential to outperform the latter with more sophisticated than uniform data models for arithmetic coder. The results are presented for grayscale and colour coding of 12 standard test images.

Signal processing techniques for ultrasonic inspection

This thesis discusses the need for nondestructive testing and highlights some of the limitations in present day techniques. Special interest has been given to ultrasonic examination techniques and the problems encountered when they are applied to thick welded plates. Some suggestions are given using signal processing methods. Chapter 2 treats the need for nondestructive testing as seen in the light of economy and safety. A short review of present day techniques in nondestructive testing is also given. The special problems using ultrasonic techniques for welded structures is discussed in Chapter 3 with some examples of elastic wave propagation in welded steel. The limitations in applying sophisticated signal processing techniques to ultrasonic NDT~ mainly found in the transducers generating or receiving the ultrasound. Chapter 4 deals with the different transducers used. One of the difficulties with ultrasonic testing is the interpretation of the signals encountered. Similar problems might be found with SONAR/RADAR techniques and Chapter 5 draws some analogies between SONAR/RADAR and ultrasonic nondestructive testing. This chapter also includes a discussion on some on the techniques used in signal processing in general. A special signal processing technique found useful is cross-correlation detection and this technique is treated in Chapter 6. Electronic digital compute.rs have made signal processing techniques easier to implement -Chapter 7 discusses the use of digital computers in ultrasonic NDT. Experimental equipment used to test cross-correlation detection of ultrasonic signals is described in Chapter 8. Chapter 9 summarises the conclusions drawn during this investigation.

Colour image coding with matching pursuit in the spatio-frequency domain

We present and evaluate a novel idea for scalable lossy colour image coding with Matching Pursuit (MP) performed in a transform domain. The idea is to exploit correlations in RGB colour space between image subbands after wavelet transformation rather than in the spatial domain. We propose a simple quantisation and coding scheme of colour MP decomposition based on Run Length Encoding (RLE) which can achieve comparable performance to JPEG 2000 even though the latter utilises careful data modelling at the coding stage. Thus, the obtained image representation has the potential to outperform JPEG 2000 with a more sophisticated coding algorithm.

Novel approaches in nonlinear optical fibre-based signal processing

All-optical data processing is expected to play a major role in future optical communications. Nonlinear effects in optical fibres have many attractive features and a great, not yet fully explored potential in optical signal processing. Here, we overview our recent advances in developing novel techniques and approaches to all-optical processing based on optical fibre nonlinearities.

All-optical nonlinear fibre signal processing

All-optical technologies for data processing and signal manipulation are expected to play a major role in future optical communications. Nonlinear phenomena occurring in optical fibre have many attractive features and great, but not yet fully exploited potential in optical signal processing. Here, we overview our recent results and advances in developing novel photonic techniques and approaches to all-optical processing based on fibre nonlinearities. Amongst other topics, we will discuss phase-preserving optical 2R regeneration, the possibility of using parabolic/flat-top pulses for optical signal processing and regeneration, and nonlinear optical pulse shaping. A method for passive nonlinear pulse shaping based on pulse pre-chirping and propagation in a normally dispersive fibre will be presented. The approach provides a simple way of generating various temporal waveforms of fundamental and practical interest. Particular emphasis will be given to the formation and characterization of pulses with a triangular intensity profile. A new technique of doubling/copying optical pulses in both the frequency and time domains using triangular-shaped pulses will be also introduced.

Nonlinearly generated advanced pulse waveforms for optical signal processing

Nonlinear phenomena occurring in optical fibres have many attractive features and great, but not yet fully explored potential in signal processing. Here, we review recent progress on the use of fibre nonlinearities for the generation and shaping of optical pulses, and on the applications of advanced pulse waveforms in all-optical signal processing. Among other topics, we will discuss ultrahigh repetition-rate pulse sources, the generation of parabolic-shaped pulses in active and passive fibres, the generation of pulses with triangular temporal profiles, and coherent supercontinuum sources. The signal processing applications will span optical regeneration, linear distortion compensation, optical decision at the receiver in optical communication systems, spectral and temporal signal doubling, and frequency conversion. © 2012 IEEE.

Recent developments in all-optical nonlinear signal processing for fiber-optic communications

All-optical data processing is expected to play a major role in future optical communications. Nonlinear effects in optical fibers have attractive applications in optical signal processing. In this paper, we review our recent advances in developing all-optical processing techniques at high speed based on optical fiber nonlinearities.

Nonlinearly generated advanced pulse waveforms for optical signal processing

Nonlinear phenomena occurring in optical fibres have many attractive features and great, but not yet fully explored potential in signal processing. Here, we review recent progress on the use of fibre nonlinearities for the generation and shaping of optical pulses, and on the applications of advanced pulse waveforms in all-optical signal processing. Among other topics, we will discuss ultrahigh repetition-rate pulse sources, the generation of parabolic-shaped pulses in active and passive fibres, the generation of pulses with triangular temporal profiles, and coherent supercontinuum sources. The signal processing applications will span optical regeneration, linear distortion compensation, optical decision at the receiver in optical communication systems, spectral and temporal signal doubling, and frequency conversion. © 2012 IEEE.

Novel approaches in nonlinear optical fibre-based signal processing

All-optical data processing is expected to play a major role in future optical communications. Nonlinear effects in optical fibres have many attractive features and a great, not yet fully explored potential in optical signal processing. Here, we overview our recent advances in developing novel techniques and approaches to all-optical processing based on optical fibre nonlinearities.