Pulse-Shape Analysis of PDM-QPSK Modulation Formats for 100 and 200 Gb/s DWDM transmissions

Advanced optical modulation format polarization-division multiplexed quadrature phase shift keying (PDM-QPSK) has become a key ingredient in the design of 100 and 200-Gb/s dense wavelength-division multiplexed (DWDM) networks. The performance of this format varies according to the shape of the pulses employed by the optical carrier: non-return to zero (NRZ), return to zero (RZ) or carrier-suppressed return to zero (CSRZ). In this paper we analyze the tolerance of PDM-QPSK to linear and nonlinear optical impairments: amplified spontaneous emission (ASE) noise, crosstalk, distortion by optical filtering, chromatic dispersion (CD), polarization mode dispersion (PMD) and fiber Kerr nonlinearities. RZ formats with a low duty cycle value reduce pulse-to-pulse interaction obtaining a higher tolerance to CD, PMD and intrachannel nonlinearities.

Design of a Two-Phase buck converter with fourth-order output filter for envelope amplifiers of limited bandwidth

The use of techniques such as envelope tracking (ET) and envelope elimination and restoration (EER) can improve the efficiency of radio frequency power amplifiers (RFPA). In both cases, high-bandwidth DC/DC converters called envelope amplifiers (EA) are used to modulate the supply voltage of the RFPA. This paper addresses the analysis and design of a modified two-phase Buck converter optimized to operate as EA. The effects of multiphase operation on the tracking capabilities are analyzed. The use of a fourth-order output filter is proposed to increase the attenuation of the harmonics generated by the PWM operation, thus allowing a reduction of the ratio between the switching frequency and the converter bandwidth. The design of the output filter is addressed considering envelope tracking accuracy and distortion caused by the side bands arising from the nonlinear modulation process. Finally, the proposed analysis and design methods are supported by simulation results, as well as demonstrated by experiments obtained using two 100-W, 10-MHz, two-phase Buck EAs capable of accurately tracking a 1.5-MHz bandwidth OFDM signal.

Contribución a la caracterización espacial de canales con sistemas MIMO-OFDM en la banda de 2,45 Ghz

La tecnología de múltiples antenas ha evolucionado para dar soporte a los actuales y futuros sistemas de comunicaciones inalámbricas en su afán por proporcionar la calidad de señal y las altas tasas de transmisión que demandan los nuevos servicios de voz, datos y multimedia. Sin embargo, es fundamental comprender las características espaciales del canal radio, ya que son las características del propio canal lo que limita en gran medida las prestaciones de los sistemas de comunicación actuales. Por ello surge la necesidad de estudiar la estructura espacial del canal de propagación para poder diseñar, evaluar e implementar de forma más eficiente tecnologías multiantena en los actuales y futuros sistemas de comunicación inalámbrica. Las tecnologías multiantena denominadas antenas inteligentes y MIMO han generado un gran interés en el área de comunicaciones inalámbricas, por ejemplo los sistemas de telefonía celular o más recientemente en las redes WLAN (Wireless Local Area Network), principalmente por la mejora que proporcionan en la calidad de las señales y en la tasa de transmisión de datos, respectivamente. Las ventajas de estas tecnologías se fundamentan en el uso de la dimensión espacial para obtener ganancia por diversidad espacial, como ya sucediera con las tecnologías FDMA (Frequency Division Multiplexing Access), TDMA (Time Division Multiplexing Access) y CDMA (Code Division Multiplexing Access) para obtener diversidad en las dimensiones de frecuencia, tiempo y código, respectivamente. Esta Tesis se centra en estudiar las características espaciales del canal con sistemas de múltiples antenas mediante la estimación de los perfiles de ángulos de llegada (DoA, Direction-of- Arrival) considerando esquemas de diversidad en espacio, polarización y frecuencia. Como primer paso se realiza una revisión de los sistemas con antenas inteligentes y los sistemas MIMO, describiendo con detalle la base matemática que sustenta las prestaciones ofrecidas por estos sistemas. Posteriormente se aportan distintos estudios sobre la estimación de los perfiles de DoA de canales radio con sistemas multiantena evaluando distintos aspectos de antenas, algoritmos de estimación, esquemas de polarización, campo lejano y campo cercano de las fuentes. Así mismo, se presenta un prototipo de medida MIMO-OFDM-SPAA3D en la banda ISM (Industrial, Scientific and Medical) de 2,45 Ghz, el cual está preparado para caracterizar experimentalmente el rendimiento de los sistemas MIMO, y para caracterizar espacialmente canales de propagación, considerando los esquemas de diversidad espacial, por polarización y frecuencia. Los estudios aportados se describen a continuación. Los sistemas de antenas inteligentes dependen en gran medida de la posición de los usuarios. Estos sistemas están equipados con arrays de antenas, los cuales aportan la diversidad espacial necesaria para obtener una representación espacial fidedigna del canal radio a través de los perfiles de DoA (DoA, Direction-of-Arrival) y por tanto, la posición de las fuentes de señal. Sin embargo, los errores de fabricación de arrays así como ciertos parámetros de señal conlleva un efecto negativo en las prestaciones de estos sistemas. Por ello se plantea un modelo de señal parametrizado que permite estudiar la influencia que tienen estos factores sobre los errores de estimación de DoA, tanto en acimut como en elevación, utilizando los algoritmos de estimación de DOA más conocidos en la literatura. A partir de las curvas de error, se pueden obtener parámetros de diseño para sistemas de localización basados en arrays. En un segundo estudio se evalúan esquemas de diversidad por polarización con los sistemas multiantena para mejorar la estimación de los perfiles de DoA en canales que presentan pérdidas por despolarización. Para ello se desarrolla un modelo de señal en array con sensibilidad de polarización que toma en cuenta el campo electromagnético de ondas planas. Se realizan simulaciones MC del modelo para estudiar el efecto de la orientación de la polarización como el número de polarizaciones usadas en el transmisor como en el receptor sobre la precisión en la estimación de los perfiles de DoA observados en el receptor. Además, se presentan los perfiles DoA obtenidos en escenarios quasiestáticos de interior con un prototipo de medida MIMO 4x4 de banda estrecha en la banda de 2,45 GHz, los cuales muestran gran fidelidad con el escenario real. Para la obtención de los perfiles DoA se propone un método basado en arrays virtuales, validado con los datos de simulación y los datos experimentales. Con relación a la localización 3D de fuentes en campo cercano (zona de Fresnel), se presenta un tercer estudio para obtener con gran exactitud la estructura espacial del canal de propagación en entornos de interior controlados (en cámara anecóica) utilizando arrays virtuales. El estudio analiza la influencia del tamaño del array y el diagrama de radiación en la estimación de los parámetros de localización proponiendo, para ello, un modelo de señal basado en un vector de enfoque de onda esférico (SWSV). Al aumentar el número de antenas del array se consigue reducir el error RMS de estimación y mejorar sustancialmente la representación espacial del canal. La estimación de los parámetros de localización se lleva a cabo con un nuevo método de búsqueda multinivel adaptativo, propuesto con el fin de reducir drásticamente el tiempo de procesado que demandan otros algoritmos multivariable basados en subespacios, como el MUSIC, a costa de incrementar los requisitos de memoria. Las simulaciones del modelo arrojan resultados que son validados con resultados experimentales y comparados con el límite de Cramer Rao en términos del error cuadrático medio. La compensación del diagrama de radiación acerca sustancialmente la exactitud de estimación de la distancia al límite de Cramer Rao. Finalmente, es igual de importante la evaluación teórica como experimental de las prestaciones de los sistemas MIMO-OFDM. Por ello, se presenta el diseño e implementación de un prototipo de medida MIMO-OFDM-SPAA3D autocalibrado con sistema de posicionamiento de antena automático en la banda de 2,45 Ghz con capacidad para evaluar la capacidad de los sistemas MIMO. Además, tiene la capacidad de caracterizar espacialmente canales MIMO, incorporando para ello una etapa de autocalibración para medir la respuesta en frecuencia de los transmisores y receptores de RF, y así poder caracterizar la respuesta de fase del canal con mayor precisión. Este sistema incorpora un posicionador de antena automático 3D (SPAA3D) basado en un scanner con 3 brazos mecánicos sobre los que se desplaza un posicionador de antena de forma independiente, controlado desde un PC. Este posicionador permite obtener una gran cantidad de mediciones del canal en regiones locales, lo cual favorece la caracterización estadística de los parámetros del sistema MIMO. Con este prototipo se realizan varias campañas de medida para evaluar el canal MIMO en términos de capacidad comparando 2 esquemas de polarización y tomando en cuenta la diversidad en frecuencia aportada por la modulación OFDM en distintos escenarios. ABSTRACT Multiple-antennas technologies have been evolved to be the support of the actual and future wireless communication systems in its way to provide the high quality and high data rates required by new data, voice and data services. However, it is important to understand the behavior of the spatial characteristics of the radio channel, since the channel by itself limits the performance of the actual wireless communications systems. This drawback raises the need to understand the spatial structure of the propagation channel in order to design, assess, and develop more efficient multiantenna technologies for the actual and future wireless communications systems. Multiantenna technologies such as ‘Smart Antennas’ and MIMO systems have generated great interest in the field of wireless communications, i.e. cellular communications systems and more recently WLAN (Wireless Local Area Networks), mainly because the higher quality and the high data rate they are able to provide. Their technological benefits are based on the exploitation of the spatial diversity provided by the use of multiple antennas as happened in the past with some multiaccess technologies such as FDMA (Frequency Division Multiplexing Access), TDMA (Time Division Multiplexing Access), and CDMA (Code Division Multiplexing Access), which give diversity in the domains of frequency, time and code, respectively. This Thesis is mainly focus to study the spatial channel characteristics using schemes of multiple antennas considering several diversity schemes such as space, polarization, and frequency. The spatial characteristics will be study in terms of the direction-of-arrival profiles viewed at the receiver side of the radio link. The first step is to do a review of the smart antennas and MIMO systems technologies highlighting their advantages and drawbacks from a mathematical point of view. In the second step, a set of studies concerning the spatial characterization of the radio channel through the DoA profiles are addressed. The performance of several DoA estimation methods is assessed considering several aspects regarding antenna array structure, polarization diversity, and far-field and near-field conditions. Most of the results of these studies come from simulations of data models and measurements with real multiantena prototypes. In the same way, having understand the importance of validate the theoretical data models with experimental results, a 2,4 GHz MIMO-OFDM-SPAA2D prototype is presented. This prototype is intended for evaluating MIMO-OFDM capacity in indoor and outdoor scenarios, characterize the spatial structure of radio channels, assess several diversity schemes such as polarization, space, and frequency diversity, among others aspects. The studies reported are briefly described below. As is stated in Chapter two, the determination of user position is a fundamental task to be resolved for the smart antenna systems. As these systems are equipped with antenna arrays, they can provide the enough spatial diversity to accurately draw the spatial characterization of the radio channel through the DoA profiles, and therefore the source location. However, certain real implementation factors related to antenna errors, signals, and receivers will certainly reduce the performance of such direction finding systems. In that sense, a parameterized narrowband signal model is proposed to evaluate the influence of these factors in the location parameter estimation through extensive MC simulations. The results obtained from several DoA algorithms may be useful to extract some parameter design for directing finding systems based on arrays. The second study goes through the importance that polarization schemes can have for estimating far-field DoA profiles in radio channels, particularly for scenarios that may introduce polarization losses. For this purpose, a narrowband signal model with polarization sensibility is developed to conduct an analysis of several polarization schemes at transmitter (TX) and receiver (RX) through extensive MC simulations. In addition, spatial characterization of quasistatic indoor scenarios is also carried out using a 2.45 GHz MIMO prototype equipped with single and dual-polarized antennas. A good agreement between the measured DoA profiles with the propagation scenario is achieved. The theoretical and experimental evaluation of polarization schemes is performed using virtual arrays. In that case, a DoA estimation method is proposed based on adding an phase reference to properly track the DoA, which shows good results. In the third study, the special case of near-field source localization with virtual arrays is addressed. Most of DoA estimation algorithms are focused in far-field source localization where the radiated wavefronts are assume to be planar waves at the receive array. However, when source are located close to the array, the assumption of plane waves is no longer valid as the wavefronts exhibit a spherical behavior along the array. Thus, a faster and effective method of azimuth, elevation angles-of-arrival, and range estimation for near-field sources is proposed. The efficacy of the proposed method is evaluated with simulation and validated with measurements collected from a measurement campaign carried out in a controlled propagation environment, i.e. anechoic chamber. Moreover, the performance of the method is assessed in terms of the RMSE for several array sizes, several source positions, and taking into account the effect of radiation pattern. In general, better results are obtained with larger array and larger source distances. The effect of the antennas is included in the data model leading to more accurate results, particularly for range rather than for angle estimation. Moreover, a new multivariable searching method based on the MUSIC algorithm, called MUSA (multilevel MUSIC-based algorithm), is presented. This method is proposed to estimate the 3D location parameters in a faster way than other multivariable algorithms, such as MUSIC algorithm, at the cost of increasing the memory size. Finally, in the last chapter, a MIMO-OFDM-SPAA3D prototype is presented to experimentally evaluate different MIMO schemes regarding antennas, polarization, and frequency in different indoor and outdoor scenarios. The prototype has been developed on a Software-Defined Radio (SDR) platform. It allows taking measurements where future wireless systems will be developed. The novelty of this prototype is concerning the following 2 subsystems. The first one is the tridimensional (3D) antenna positioning system (SPAA3D) based on three linear scanners which is developed for making automatic testing possible reducing errors of the antenna array positioning. A set of software has been developed for research works such as MIMO channel characterization, MIMO capacity, OFDM synchronization, and so on. The second subsystem is the RF autocalibration module at the TX and RX. This subsystem allows to properly tracking the spatial structure of indoor and outdoor channels in terms of DoA profiles. Some results are draw regarding performance of MIMO-OFDM systems with different polarization schemes and different propagation environments.

Random-modulation cw lidar system for space-borne carbon dioxide remote sensing based on a high-brightness semiconductor laser

In this paper, we report on the progresses of the BRITESPACE Consortium in order to achieve space-borne LIDAR measurements of atmospheric carbon dioxide concentration based on an all semiconductor laser source at 1.57 ?m. The complete design of the proposed RM-CW IPDA LIDAR has been presented and described in detail. Complete descriptions of the laser module and the FSU have been presented. Two bended MOPAs, emitting at the sounding frequency of the on- and off- IPDA channels, have been proposed as the transmitter optical sources with the required high brightness. Experimental results on the bended MOPAs have been presented showing a high spectral purity and promising expectations on the high output power requirements. Finally, the RM-CW approach has been modelled and an estimation of the expected SNR for the entire system is presented. Preliminary results indicate that a CO2 retrieval precision of 1.5 ppm could be achieved with an average output power of 2 W for each channel.

Tuning of modulation spectrum parameters for voice pathology detection

Acoustic parameters are frequently used to assess the presence of pathologies in human voice. Many of them have demonstrated to be useful but in some cases its results could be optimized by selecting appropriate working margins. In this study two indices, CIL and RALA, obtained from Modulation Spectra are described and tuned using different frame lengths and frequency ranges to maximize AUC in normal to pathological voice detection. After the tuning process, AUC reaches 0.96 and 0.95 values for CIL and RALA respectively representing an improvement of 16 % and 12 % at each case respect to the typical tuning based only on frame length selection.

Factor-specific modulation of CREB-binding protein acetyltransferase activity

CREB-binding proteins (CBP) and p300 are essential transcriptional coactivators for a large number of regulated DNA-binding transcription factors, including CREB, nuclear receptors, and STATs. CBP and p300 function in part by mediating the assembly of multiprotein complexes that contain additional cofactors such as p300/CBP interacting protein (p/CIP), a member of the p160/SRC family of coactivators, and the p300/CBP associated factor p/CAF. In addition to serving as molecular scaffolds, CBP and p300 each possess intrinsic acetyltransferase activities that are required for their function as coactivators. Here we report that the adenovirus E1A protein inhibits the acetyltransferase activity of CBP on binding to the C/H3 domain, whereas binding of CREB, or a CREB/E1A fusion protein to the KIX domain, fails to inhibit CBP acetyltransferase activity. Surprisingly, p/CIP can either inhibit or stimulate CBP acetyltransferase activity depending on the specific substrate evaluated and the functional domains present in the p/CIP protein. While the CBP interaction domain of p/CIP inhibits acetylation of histones H3, H4, or high mobility group by CBP, it enhances acetylation of other substrates, such as Pit-1. These observations suggest that the acetyltransferase activities of CBP/p300 and p/CAF can be differentially modulated by factors binding to distinct regions of CBP/p300. Because these interactions are likely to result in differential effects on the coactivator functions of CBP/p300 for different classes of transcription factors, regulation of CBP/p300 acetyltransferase activity may represent a mechanism for integration of diverse signaling pathways.

Modulation of nicotinic acetylcholine receptors by strychnine

Strychnine, a potent and selective antagonist at glycine receptors, was found to inhibit muscle (α1β1γδ, α1β1γ, and α1β1δ) and neuronal (α2β2 and α2β4) nicotinic acetylcholine receptors (AcChoRs) expressed in Xenopus oocytes. Strychnine alone (up to 500 μM) did not elicit membrane currents in oocytes expressing AcChoRs, but, when applied before, concomitantly, or during superfusion of acetylcholine (AcCho), it rapidly and reversibly inhibited the current elicited by AcCho (AcCho-current). Although in the three cases the AcCho-current was reduced to the same level, its recovery was slower when the oocytes were preincubated with strychnine. The amount of AcCho-current inhibition depended on the receptor subtype, and the order of blocking potency by strychnine was α1β1γδ > α2β4 > α2β2. With the three forms of drug application, the Hill coefficient was close to one, suggesting a single site for the receptor interaction with strychnine, and this interaction appears to be noncompetitive. The inhibitory effects on muscle AcChoRs were voltage-independent, and the apparent dissociation constant for AcCho was not appreciably changed by strychnine. In contrast, the inhibitory effects on neuronal AcChoRs were voltage-dependent, with an electrical distance of ≈0.35. We conclude that strychnine regulates reversibly and noncompetitively the embryonic type of muscle AcChoR and some forms of neuronal AcChoRs. In the former case, strychnine presumably inhibits allosterically the receptor by binding at an external domain whereas, in the latter case, it blocks the open receptor-channel complex.

Modulation of an early step in the secretory machinery in hippocampal nerve terminals

In hippocampal neurons, neurotransmitter release can be regulated by protein kinase A (PKA) through a direct action on the secretory machinery. To identify the site of PKA modulation, we have taken advantage of the ability of the neurotoxin Botulinum A to cleave the synaptic protein SNAP-25. Cleavage of this protein decreases the Ca2+ responsiveness of the secretory machinery by partially uncoupling Ca2+-sensing from fusion per se. This is expressed as a shift toward higher Ca2+ levels of the Ca2+ to neurotransmitter release relationship and as a perturbation of synaptic delay under conditions where secretion induced by the Ca2+-independent secretagogue ruthenium red is unimpaired. We find that SNAP-25 cleavage also perturbs PKA-dependent modulation of secretion; facilitation of ruthenium red-evoked neurotransmitter release by the adenylyl cyclase activator forskolin is blocked completely after Botulinum toxin A action. Together with our observation that forskolin modifies the Ca2+ to neurotransmitter release relationship, our results suggest that SNAP-25 acts as a functional linker between Ca2+ detection and fusion and that PKA modulates an early step in the secretory machinery related to calcium sensing to facilitate synaptic transmission.

Solution of the pulse width modulation problem using orthogonal polynomials and Korteweg-de Vries equations

The mathematical underpinning of the pulse width modulation (PWM) technique lies in the attempt to represent “accurately” harmonic waveforms using only square forms of a fixed height. The accuracy can be measured using many norms, but the quality of the approximation of the analog signal (a harmonic form) by a digital one (simple pulses of a fixed high voltage level) requires the elimination of high order harmonics in the error term. The most important practical problem is in “accurate” reproduction of sine-wave using the same number of pulses as the number of high harmonics eliminated. We describe in this paper a complete solution of the PWM problem using Padé approximations, orthogonal polynomials, and solitons. The main result of the paper is the characterization of discrete pulses answering the general PWM problem in terms of the manifold of all rational solutions to Korteweg-de Vries equations.

Translational coupling by modulation of feedback repression in the IF3 operon of Escherichia coli

A pseudoknot formed by a long-range interaction in the mRNA of the initiation factor 3 (IF3) operon is involved in the translational repression of the gene encoding ribosomal protein L35 by another ribosomal protein, L20. The nucleotides forming the 5′ strand of the key stem of the pseudoknot are located within the gene for IF3, whereas those forming the 3′ strand are located 280 nt downstream, immediately upstream of the Shine–Dalgarno sequence of the gene for L35. Here we show that premature termination of IF3 translation at a nonsense codon introduced upstream of the pseudoknot results in a substantial enhancement of L20-mediated repression of L35 expression. Conversely, an increase of IF3 translation decreases repression. These results, in addition to an analysis of the effect of mutations in sequences forming the pseudoknot, indicate that IF3 translation decreases L20-mediated repression of L35 expression. We propose that ribosomes translating IF3 disrupt the pseudoknot and thereby attenuate repression. The result is a novel type of translational coupling, where unfolding of the pseudoknot by ribosomes translating IF3 does not increase expression of L35 directly, but alleviates its repression by L20.

Modulation of cognition-specific cortical activity by gonadal steroids: A positron-emission tomography study in women

There is considerable evidence from animal studies that gonadal steroid hormones modulate neuronal activity and affect behavior. To study this in humans directly, we used H215O positron-emission tomography to measure regional cerebral blood flow (rCBF) in young women during three pharmacologically controlled hormonal conditions spanning 4–5 months: ovarian suppression induced by the gonadotropin-releasing hormone agonist leuprolide acetate (Lupron), Lupron plus estradiol replacement, and Lupron plus progesterone replacement. Estradiol and progesterone were administered in a double-blind cross-over design. On each occasion positron-emission tomography scans were performed during (i) the Wisconsin Card Sorting Test, a neuropsychological test that physiologically activates prefrontal cortex (PFC) and an associated cortical network including inferior parietal lobule and posterior inferolateral temporal gyrus, and (ii) a no-delay matching-to-sample sensorimotor control task. During treatment with Lupron alone (i.e., with virtual absence of gonadal steroid hormones), there was marked attenuation of the typical Wisconsin Card Sorting Test activation pattern even though task performance did not change. Most strikingly, there was no rCBF increase in PFC. When either progesterone or estrogen was added to the Lupron regimen, there was normalization of the rCBF activation pattern with augmentation of the parietal and temporal foci and return of the dorsolateral PFC activation. These data directly demonstrate that the hormonal milieu modulates cognition-related neural activity in humans.

Endocrine modulation of the neurotoxicity of gp120: Implications for AIDS-related dementia complex

HIV infection often involves the development of AIDS-related dementia complex, a variety of neurologic, neuropsychologic, and neuropathologic impairments. A possible contributor to AIDS-related dementia complex is the HIV envelope glycoprotein gp120, which damages neurons via a complex glutamate receptor- and calcium-dependent cascade. We demonstrate an endocrine modulation of the deleterious effects of gp120 in primary hippocampal and cortical cultures. Specifically, we observe that gp120-induced calcium mobilization and neurotoxicity are exacerbated by glucocorticoids, the adrenal steroids secreted during stress. Importantly, this deleterious synergy can occur between gp120 and synthetic glucocorticoids (such as prednisone or dexamethasone) that are used clinically in high concentrations to treat severe cases of the Pneumocystis carinii pneumonia typical of HIV infection. Conversely, we also observe that estradiol protects neurons from the deleterious actions of gp120, reducing toxicity and calcium mobilization.

Modulation of potassium channel function by methionine oxidation and reduction

Oxidation of amino acid residues in proteins can be caused by a variety of oxidizing agents normally produced by cells. The oxidation of methionine in proteins to methionine sulfoxide is implicated in aging as well as in pathological conditions, and it is a reversible reaction mediated by a ubiquitous enzyme, peptide methionine sulfoxide reductase. The reversibility of methionine oxidation suggests that it could act as a cellular regulatory mechanism although no such in vivo activity has been demonstrated. We show here that oxidation of a methionine residue in a voltage-dependent potassium channel modulates its inactivation. When this methionine residue is oxidized to methionine sulfoxide, the inactivation is disrupted, and it is reversed by coexpression with peptide methionine sulfoxide reductase. The results suggest that oxidation and reduction of methionine could play a dynamic role in the cellular signal transduction process in a variety of systems.

The cysteine-rich frizzled domain of Frzb-1 is required and sufficient for modulation of Wnt signaling

Convincing evidence has accumulated to identify the Frizzled proteins as receptors for the Wnt growth factors. In parallel, a number of secreted frizzled-like proteins with a conserved N-terminal frizzled motif have been identified. One of these proteins, Frzb-1, binds Wnt-1 and Xwnt-8 proteins and antagonizes Xwnt-8 signaling in Xenopus embryos. Here we report that Frzb-1 blocks Wnt-1 induced cytosolic accumulation of β-catenin, a key component of the Wnt signaling pathway, in human embryonic kidney cells. Structure/function analysis reveals that complete removal of the frizzled domain of Frzb-1 abolishes the Wnt-1/Frzb-1 protein interaction and the inhibition of Wnt-1 mediated axis duplication in Xenopus embryos. In contrast, removal of the C-terminal portion of the molecule preserves both Frzb-Wnt binding and functional inhibition of Wnt signaling. Partial deletions of the Frzb-1 cysteine-rich domain maintain Wnt-1 interaction, but functional inhibition is lost. Taken together, these findings support the conclusion that the frizzled domain is necessary and sufficient for both activities. Interestingly, Frzb-1 does not block Wnt-5A signaling in a Xenopus functional assay, even though Wnt-5A coimmunoprecipitates with Frzb-1, suggesting that coimmunoprecipitation does not necessarily imply inhibition of Wnt function.

Iron regulatory protein 1 is not required for the modulation of ferritin and transferrin receptor expression by iron in a murine pro-B lymphocyte cell line

Iron regulatory proteins (IRPs) are cytoplasmic RNA binding proteins that are central components of a sensory and regulatory network that modulates vertebrate iron homeostasis. IRPs regulate iron metabolism by binding to iron responsive element(s) (IREs) in the 5′ or 3′ untranslated region of ferritin or transferrin receptor (TfR) mRNAs. Two IRPs, IRP1 and IRP2, have been identified previously. IRP1 exhibits two mutually exclusive functions as an RNA binding protein or as the cytosolic isoform of aconitase. We demonstrate that the Ba/F3 family of murine pro-B lymphocytes represents the first example of a mammalian cell line that fails to express IRP1 protein or mRNA. First, all of the IRE binding activity in Ba/F3-gp55 cells is attributable to IRP2. Second, synthesis of IRP2, but not of IRP1, is detectable in Ba/F3-gp55 cells. Third, the Ba/F3 family of cells express IRP2 mRNA at a level similar to other murine cell lines, but IRP1 mRNA is not detectable. In the Ba/F3 family of cells, alterations in iron status modulated ferritin biosynthesis and TfR mRNA level over as much as a 20- and 14-fold range, respectively. We conclude that IRP1 is not essential for regulation of ferritin or TfR expression by iron and that IRP2 can act as the sole IRE-dependent mediator of cellular iron homeostasis.