Coupling the phosphotransferase system and the methyl-accepting chemotaxis protein-dependent chemotaxis signaling pathways of Escherichia coli.

Chemotactic responses in Escherichia coli are typically mediated by transmembrane receptors that monitor chemoeffector levels with periplasmic binding domains and communicate with the flagellar motors through two cytoplasmic proteins, CheA and CheY. CheA autophosphorylates and then donates its phosphate to CheY, which in turn controls flagellar rotation. E. coli also exhibits chemotactic responses to substrates that are transported by the phosphoenolpyruvate (PEP)-dependent carbohydrate phosphotransferase system (PTS). Unlike conventional chemoreception, PTS substrates are sensed during their uptake and concomitant phosphorylation by the cell. The phosphoryl groups are transferred from PEP to the carbohydrates through two common intermediates, enzyme I (EI) and phosphohistidine carrier protein (HPr), and then to sugar-specific enzymes II. We found that in mutant strains HPr-like proteins could substitute for HPr in transport but did not mediate chemotactic signaling. In in vitro assays, these proteins exhibited reduced phosphotransfer rates from EI, indicating that the phosphorylation state of EI might link the PTS phospho-relay to the flagellar signaling pathway. Tests with purified proteins revealed that unphosphorylated EI inhibited CheA autophosphorylation, whereas phosphorylated EI did not. These findings suggest the following model for signal transduction in PTS-dependent chemotaxis. During uptake of a PTS carbohydrate, EI is dephosphorylated more rapidly by HPr than it is phosphorylated at the expense of PEP. Consequently, unphosphorylated EI builds up and inhibits CheA autophosphorylation. This slows the flow of phosphates to CheY, eliciting an up-gradient swimming response by the cell.

Novel particulate BCG-loaded delivery system for mucosal immunization against tuberculosis

Tese de doutoramento, Farmácia (Tecnologia Farmacêutica), Universidade de Lisboa, Faculdade de Farmácia, 2016

Polymorphic CAG repeat length in the androgen receptor gene and association with neurodegeneration in a heterozygous female carrier of Kennedy's disease

Kennedy's disease (spinobulbar muscular atrophy) is an X-linked form of motor neuron disease affecting adult males carrying a CAG trinucleotide repeat expansion within the androgen receptor gene. While expression of Kennedy's disease is thought to be confined to males carrying the causative mutation, subclinical manifestations have been reported in a few female carriers of the disease. The reasons that females are protected from the disease are not clear, especially given that all other diseases caused by CAG expansions display dominant expression. In the current study, we report the identification of a heterozygote female carrying the Kennedy's disease mutation who was clinically diagnosed with motor neuron disease. We describe analysis of CAG repeat number in this individual as well as 33 relatives within the pedigree, including two male carriers of the Kennedy's mutation. The female heterozygote carried one expanded allele of the androgen receptor gene with CAG repeats numbering in the Kennedy's disease range (44 CAGs), with the normal allele numbering in the upper-normal range (28 CAGs). The subject has two sons, one of whom carries the mutant allele of the gene and has been clinically diagnosed with Kennedy's disease, whilst the other son carries the second allele of the gene with CAGs numbering in the upper normal range and displays a normal phenotype. This coexistence of motor neuron disease and the presence of one expanded allele and one allele at the upper limit of the normal range may be a coincidence. However, we hypothesize that the expression of the Kennedy's disease mutation combined with a second allele with a large but normal CAG repeat sequence may have contributed to the motor neuron degeneration displayed in the heterozygote female and discuss the possible reasons for phenotypic expression in particular individuals.

Towards an improved ocular drug delivery system

The ultimate aim of this project was to design new biomaterials which will improve the efficiency of ocular drug delivery systems. Initially, it was necessary to review the information available on the nature of the tear fluid and its relationship with the eye. An extensive survey of the relevant literature was made. There is a common belief in the literature that the ocular glycoprotein, mucin, plays an important role in tear film stability, and furthermore, that it exists as an adherent layer covering the corneal surface. If this belief is true, the muco-corneal interaction provides the ideal basis for the development of sustained release drug delivery. Preliminary investigations were made to assess the ability of mucin to adhere to polymer surfaces. The intention was to develop a synthetic model which would mimic the supposed corneal/mucin interaction. Analytical procedures included the use of microscopy (phase contrast and fluorescence), fluorophotometry, and mucin-staining dyes. Additionally, the physical properties of tears and tear models were assessed under conditions mimicking those of the preocular environment, using rheological and tensiometric techniques. The wetting abilities of these tear models and opthalmic formulations were also investigated. Tissue culture techniques were employed to enable the surface properties of the corneal surface to be studied by means of cultured corneal cells. The results of these investigations enabled the calculation of interfacial and surface characteristics of tears, tear models, and the corneal surface. Over all, this work cast doubt on the accepted relationship of mucin with the cornea. A corneal surface model was designed, on the basis of the information obtained during this project, which would possess similar surface chemical properties (i.e. would be biomimetic) to the more complex original. This model, together with the information gained on the properties of tears and solutions intended for ocular instillation, could be valuable in the design of drug formulations with enhanced ocular retention times. Furthermore, the model itself may form the basis for the design of an effective drug-carrier.

A low-cost remote sensing system for agricultural applications

This research develops a low cost remote sensing system for use in agricultural applications. The important features of the system are that it monitors the near infrared and it incorporates position and attitude measuring equipment allowing for geo-rectified images to be produced without the use of ground control points. The equipment is designed to be hand held and hence requires no structural modification to the aircraft. The portable remote sensing system consists of an inertia measurement unit (IMU), which is accelerometer based, a low-cost GPS device and a small format false colour composite digital camera. The total cost of producing such a system is below GBP 3000, which is far cheaper than equivalent existing systems. The design of the portable remote sensing device has eliminated bore sight misalignment errors from the direct geo-referencing process. A new processing technique has been introduced for the data obtained from these low-cost devices, and it is found that using this technique the image can be matched (overlaid) onto Ordnance Survey Master Maps at an accuracy compatible with precision agriculture requirements. The direct geo-referencing has also been improved by introducing an algorithm capable of correcting oblique images directly. This algorithm alters the pixels value, hence it is advised that image analysis is performed before image georectification. The drawback of this research is that the low-cost GPS device experienced bad checksum errors, which resulted in missing data. The Wide Area Augmented System (WAAS) correction could not be employed because the satellites could not be locked onto whilst flying. The best GPS data were obtained from the Garmin eTrex (15 m kinematic and 2 m static) instruments which have a highsensitivity receiver with good lock on capability. The limitation of this GPS device is the inability to effectively receive the P-Code wavelength, which is needed to gain the best accuracy when undertaking differential GPS processing. Pairing the carrier phase L1 with the pseudorange C/A-Code received, in order to determine the image coordinates by the differential technique, is still under investigation. To improve the position accuracy, it is recommended that a GPS base station should be established near the survey area, instead of using a permanent GPS base station established by the Ordnance Survey.

Pattern integration in the normal and abnormal human visual system

The processing conducted by the visual system requires the combination of signals that are detected at different locations in the visual field. The processes by which these signals are combined are explored here using psychophysical experiments and computer modelling. Most of the work presented in this thesis is concerned with the summation of contrast over space at detection threshold. Previous investigations of this sort have been confounded by the inhomogeneity in contrast sensitivity across the visual field. Experiments performed in this thesis find that the decline in log contrast sensitivity with eccentricity is bilinear, with an initial steep fall-off followed by a shallower decline. This decline is scale-invariant for spatial frequencies of 0.7 to 4 c/deg. A detailed map of the inhomogeneity is developed, and applied to area summation experiments both by incorporating it into models of the visual system and by using it to compensate stimuli in order to factor out the effects of the inhomogeneity. The results of these area summation experiments show that the summation of contrast over area is spatially extensive (occurring over 33 stimulus carrier cycles), and that summation behaviour is the same in the fovea, parafovea, and periphery. Summation occurs according to a fourth-root summation rule, consistent with a “noisy energy” model. This work is extended to investigate the visual deficit in amblyopia, finding that area summation is normal in amblyopic observers. Finally, the methods used to study the summation of threshold contrast over area are adapted to investigate the integration of coherent orientation signals in a texture. The results of this study are described by a two-stage model, with a mandatory local combination stage followed by flexible global pooling of these local outputs. In each study, the results suggest a more extensive combination of signals in vision than has been previously understood.

10 Mb/s visible light transmission system using a polymer light-emitting diode with orthogonal frequency division multiplexing

We present a newly designed polymer light-emitting diode with a bandwidth of ∼350 kHz for high-speed visible light communications. Using this new polymer light-emitting diode as a transmitter, we have achieved a record transmission speed of 10 Mb/s for a polymer light-emitting diode-based optical communication system with an orthogonal frequency division multiplexing technique, matching the performance of single carrier formats using multitap equalization. For achieving such a high data-rate, a power pre-emphasis technique was adopted. © 2014 Optical Society of America.

112Gbit/s RF assisted dual-carrier DP-16-QAM in installed and new build WDM applications

The performance of a 112Gbit/s dual-carrier DP-16-QAM channel in various WDM configurations is characterized. Variations of the dispersion map, ROADM count and system length are experimentally evaluated and compared with numerical simulation. © 2012 OSA.

Analytical BER performance in differential n-PSK coherent transmission system influenced by equalization enhanced phase noise

Long-haul high speed optical transmission systems are significantly distorted by the interplay between the electronic chromatic dispersion (CD) equalization and the local oscillator (LO) laser phase noise, which leads to an effect of equalization enhanced phase noise (EEPN). The EEPN degrades the performance of optical communication systems severely with the increment of fiber dispersion, LO laser linewidth, symbol rate, and modulation format. In this paper, we present an analytical model for evaluating the performance of bit-error-rate (BER) versus signal-to-noise ratio (SNR) in the n-level phase shift keying (n-PSK) coherent transmission system employing differential carrier phase estimation (CPE), where the influence of EEPN is considered. Theoretical results based on this model have been investigated for the differential quadrature phase shift keying (DQPSK), the differential 8-PSK (D8PSK), and the differential 16-PSK (D16PSK) coherent transmission systems. The influence of EEPN on the BER performance in term of the fiber dispersion, the LO phase noise, the symbol rate, and the modulation format are analyzed in detail. The BER behaviors based on this analytical model achieve a good agreement with previously reported BER floors influenced by EEPN. Further simulations have also been carried out in the differential CPE considering EEPN. The results indicate that this analytical model can give an accurate prediction for the DQPSK system, and a leading-order approximation for the D8PSK and the D16PSK systems.

Experimental full duplex simultaneous transmission of LTE over a DWDM directly modulated RoF system

In this paper, we experimentally demonstrate the seamless integration of full duplex system frequency division duplex (FDD) long-term evolution (LTE) technology with radio over fiber (RoF) for eNodeB (eNB) coverage extension. LTE is composed of quadrature phase-shift keying (QPSK), 16-quadrature amplitude modulation (16-QAM) and 64-QAM, modulated onto orthogonal frequency division multiplexing (OFDM) and single-carrier-frequency division multiplexing for downlink (DL) and uplink (UL) transmissions, respectively. The RoF system is composed of dedicated directly modulated lasers for DL and UL with dense wavelength division multiplexing (DWDM) for instantaneous connections and for Rayleigh backscattering and nonlinear interference mitigation. DL and UL signals have varying carrier frequencies and are categorized as broad frequency spacing (BFS), intermediate frequency spacing (IFS), and narrow frequency spacing (NFS). The adjacent channel leakage ratio (ACLR) for DL and UL with 64-QAM are similar for all frequency spacings while cross talk is observed for NFS. For the best case scenario for DL and UL transmissions we achieve error vector magnitude (EVM) values of ~2.30%, ~2.33%, and ~2.39% for QPSK, 16-QAM, and 64-QAM, respectively, while for the worst case scenario with a NFS EVM is increased by 0.40% for all schemes. © 2009-2012 OSA.

Peak -to -average power ratio reduced parallel interference cancellation Multicarrier-Code Division Multiple Access system with anti -interference property

Orthogonal Frequency-Division Multiplexing (OFDM) has been proved to be a promising technology that enables the transmission of higher data rate. Multicarrier Code-Division Multiple Access (MC-CDMA) is a transmission technique which combines the advantages of both OFDM and Code-Division Multiplexing Access (CDMA), so as to allow high transmission rates over severe time-dispersive multi-path channels without the need of a complex receiver implementation. Also MC-CDMA exploits frequency diversity via the different subcarriers, and therefore allows the high code rates systems to achieve good Bit Error Rate (BER) performances. Furthermore, the spreading in the frequency domain makes the time synchronization requirement much lower than traditional direct sequence CDMA schemes. There are still some problems when we use MC-CDMA. One is the high Peak-to-Average Power Ratio (PAPR) of the transmit signal. High PAPR leads to nonlinear distortion of the amplifier and results in inter-carrier self-interference plus out-of-band radiation. On the other hand, suppressing the Multiple Access Interference (MAI) is another crucial problem in the MC-CDMA system. Imperfect cross-correlation characteristics of the spreading codes and the multipath fading destroy the orthogonality among the users, and then cause MAI, which produces serious BER degradation in the system. Moreover, in uplink system the received signals at a base station are always asynchronous. This also destroys the orthogonality among the users, and hence, generates MAI which degrades the system performance. Besides those two problems, the interference should always be considered seriously for any communication system. In this dissertation, we design a novel MC-CDMA system, which has low PAPR and mitigated MAI. The new Semi-blind channel estimation and multi-user data detection based on Parallel Interference Cancellation (PIC) have been applied in the system. The Low Density Parity Codes (LDPC) has also been introduced into the system to improve the performance. Different interference models are analyzed in multi-carrier communication systems and then the effective interference suppression for MC-CDMA systems is employed in this dissertation. The experimental results indicate that our system not only significantly reduces the PAPR and MAI but also effectively suppresses the outside interference with low complexity. Finally, we present a practical cognitive application of the proposed system over the software defined radio platform.

Peak-to-Average Power Ratio Reduced Parallel Interference Cancellation Multicarrier-Code Division Multiple Access System with Anti-Interference Property

Orthogonal Frequency-Division Multiplexing (OFDM) has been proved to be a promising technology that enables the transmission of higher data rate. Multicarrier Code-Division Multiple Access (MC-CDMA) is a transmission technique which combines the advantages of both OFDM and Code-Division Multiplexing Access (CDMA), so as to allow high transmission rates over severe time-dispersive multi-path channels without the need of a complex receiver implementation. Also MC-CDMA exploits frequency diversity via the different subcarriers, and therefore allows the high code rates systems to achieve good Bit Error Rate (BER) performances. Furthermore, the spreading in the frequency domain makes the time synchronization requirement much lower than traditional direct sequence CDMA schemes. There are still some problems when we use MC-CDMA. One is the high Peak-to-Average Power Ratio (PAPR) of the transmit signal. High PAPR leads to nonlinear distortion of the amplifier and results in inter-carrier self-interference plus out-of-band radiation. On the other hand, suppressing the Multiple Access Interference (MAI) is another crucial problem in the MC-CDMA system. Imperfect cross-correlation characteristics of the spreading codes and the multipath fading destroy the orthogonality among the users, and then cause MAI, which produces serious BER degradation in the system. Moreover, in uplink system the received signals at a base station are always asynchronous. This also destroys the orthogonality among the users, and hence, generates MAI which degrades the system performance. Besides those two problems, the interference should always be considered seriously for any communication system. In this dissertation, we design a novel MC-CDMA system, which has low PAPR and mitigated MAI. The new Semi-blind channel estimation and multi-user data detection based on Parallel Interference Cancellation (PIC) have been applied in the system. The Low Density Parity Codes (LDPC) has also been introduced into the system to improve the performance. Different interference models are analyzed in multi-carrier communication systems and then the effective interference suppression for MC-CDMA systems is employed in this dissertation. The experimental results indicate that our system not only significantly reduces the PAPR and MAI but also effectively suppresses the outside interference with low complexity. Finally, we present a practical cognitive application of the proposed system over the software defined radio platform.