Application and modeling of GaN FET in 1MHz large signal bandwidth power supply for radio frequency power amplifier

In this paper, implementation and testing of non- commercial GaN HEMT in a simple buck converter for envelope amplifier in ET and EER transmission techn iques has been done. Comparing to the prototypes with commercially available EPC1014 and 1015 GaN HEMTs, experimentally demonstrated power supply provided better thermal management and increased the switching frequency up to 25MHz. 64QAM signal with 1MHz of large signal bandw idth and 10.5dB of Peak to Average Power Ratio was gener ated, using the switching frequency of 20MHz. The obtaine defficiency was 38% including the driving circuit an d the total losses breakdown showed that switching power losses in the HEMT are the dominant ones. In addition to this, some basic physical modeling has been done, in order to provide an insight on the correlation between the electrical characteristics of the GaN HEMT and physical design parameters. This is the first step in the optimization of the HEMT design for this particular application.

Wide bandwidth envelope trackers with reduced switching frequency for RF power amplifier

El requerimiento de proveer alta frecuencia de datos en los modernos sistema de comunicación inalámbricos resulta en complejas señales moduladas de radio-frequencia (RF) con un gran ancho de banda y alto ratio pico-promedio (PAPR). Para garantizar la linealidad del comportamiento, los amplificadores lineales de potencia comunes funcionan típicamente entre 4 y 10 dB de back-o_ desde la máxima potencia de salida, ocasionando una baja eficiencia del sistema. La eliminación y restauración de la evolvente (EER) y el seguimiento de la evolvente (ET) son dos prometedoras técnicas para resolver el problema de la eficiencia. Tanto en EER como en ET, es complicado diseñar un amplificador de potencia que sea eficiente para señales de RF de alto ancho de banda y alto PAPR. Una propuesta común para los amplificadores de potencia es incluir un convertidor de potencia de muy alta eficiencia operando a frecuencias más altas que el ancho de banda de la señal RF. En este caso, la potencia perdida del convertidor ocasionado por la alta frecuencia desaconseja su práctica cuando el ancho de banda es muy alto. La solución a este problema es el enfoque de esta disertación que presenta dos arquitecturas de amplificador evolvente: convertidor híbrido-serie con una técnica de evolvente lenta y un convertidor multinivel basado en un convertidor reductor multifase con control de tiempo mínimo. En la primera arquitectura, una topología híbrida está compuesta de una convertidor reductor conmutado y un regulador lineal en serie que trabajan juntos para ajustar la tensión de salida para seguir a la evolvente con precisión. Un algoritmo de generación de una evolvente lenta crea una forma de onda con una pendiente limitada que es menor que la pendiente máxima de la evolvente original. La salida del convertidor reductor sigue esa forma de onda en vez de la evolvente original usando una menor frecuencia de conmutación, porque la forma de onda no sólo tiene una pendiente reducida sino también un menor ancho de banda. De esta forma, el regulador lineal se usa para filtrar la forma de onda tiene una pérdida de potencia adicional. Dependiendo de cuánto se puede reducir la pendiente de la evolvente para producir la forma de onda, existe un trade-off entre la pérdida de potencia del convertidor reductor relacionada con la frecuencia de conmutación y el regulador lineal. El punto óptimo referido a la menor pérdida de potencia total del amplificador de evolvente es capaz de identificarse con la ayuda de modelo preciso de pérdidas que es una combinación de modelos comportamentales y analíticos de pérdidas. Además, se analiza el efecto en la respuesta del filtro de salida del convertidor reductor. Un filtro de dampeo paralelo extra es necesario para eliminar la oscilación resonante del filtro de salida porque el convertidor reductor opera en lazo abierto. La segunda arquitectura es un amplificador de evolvente de seguimiento de tensión multinivel. Al contrario que los convertidores que usan multi-fuentes, un convertidor reductor multifase se emplea para generar la tensión multinivel. En régimen permanente, el convertidor reductor opera en puntos del ciclo de trabajo con cancelación completa del rizado. El número de niveles de tensión es igual al número de fases de acuerdo a las características del entrelazamiento del convertidor reductor. En la transición, un control de tiempo mínimo (MTC) para convertidores multifase es novedosamente propuesto y desarrollado para cambiar la tensión de salida del convertidor reductor entre diferentes niveles. A diferencia de controles convencionales de tiempo mínimo para convertidores multifase con inductancia equivalente, el propuesto MTC considera el rizado de corriente por cada fase basado en un desfase fijo que resulta en diferentes esquemas de control entre las fases. La ventaja de este control es que todas las corrientes vuelven a su fase en régimen permanente después de la transición para que la siguiente transición pueda empezar muy pronto, lo que es muy favorable para la aplicación de seguimiento de tensión multinivel. Además, el control es independiente de la carga y no es afectado por corrientes de fase desbalanceadas. Al igual que en la primera arquitectura, hay una etapa lineal con la misma función, conectada en serie con el convertidor reductor multifase. Dado que tanto el régimen permanente como el estado de transición del convertidor no están fuertemente relacionados con la frecuencia de conmutación, la frecuencia de conmutación puede ser reducida para el alto ancho de banda de la evolvente, la cual es la principal consideración de esta arquitectura. La optimización de la segunda arquitectura para más alto anchos de banda de la evolvente es presentada incluyendo el diseño del filtro de salida, la frecuencia de conmutación y el número de fases. El área de diseño del filtro está restringido por la transición rápida y el mínimo pulso del hardware. La rápida transición necesita un filtro pequeño pero la limitación del pulso mínimo del hardware lleva el diseño en el sentido contrario. La frecuencia de conmutación del convertidor afecta principalmente a la limitación del mínimo pulso y a las pérdidas de potencia. Con una menor frecuencia de conmutación, el ancho de pulso en la transición es más pequeño. El número de fases relativo a la aplicación específica puede ser optimizado en términos de la eficiencia global. Otro aspecto de la optimización es mejorar la estrategia de control. La transición permite seguir algunas partes de la evolvente que son más rápidas de lo que el hardware puede soportar al precio de complejidad. El nuevo método de sincronización de la transición incrementa la frecuencia de la transición, permitiendo que la tensión multinivel esté más cerca de la evolvente. Ambas estrategias permiten que el convertidor pueda seguir una evolvente con un ancho de banda más alto que la limitación de la etapa de potencia. El modelo de pérdidas del amplificador de evolvente se ha detallado y validado mediante medidas. El mecanismo de pérdidas de potencia del convertidor reductor tiene que incluir las transiciones en tiempo real, lo cual es diferente del clásico modelos de pérdidas de un convertidor reductor síncrono. Este modelo estima la eficiencia del sistema y juega un papel muy importante en el proceso de optimización. Finalmente, la segunda arquitectura del amplificador de evolvente se integra con el amplificador de clase F. La medida del sistema EER prueba el ahorro de energía con el amplificador de evolvente propuesto sin perjudicar la linealidad del sistema. ABSTRACT The requirement of delivering high data rates in modern wireless communication systems results in complex modulated RF signals with wide bandwidth and high peak-to-average ratio (PAPR). In order to guarantee the linearity performance, the conventional linear power amplifiers typically work at 4 to 10 dB back-off from the maximum output power, leading to low system efficiency. The envelope elimination and restoration (EER) and envelope tracking (ET) are two promising techniques to overcome the efficiency problem. In both EER and ET, it is challenging to design efficient envelope amplifier for wide bandwidth and high PAPR RF signals. An usual approach for envelope amplifier includes a high-efficiency switching power converter operating at a frequency higher than the RF signal's bandwidth. In this case, the power loss of converter caused by high switching operation becomes unbearable for system efficiency when signal bandwidth is very wide. The solution of this problem is the focus of this dissertation that presents two architectures of envelope amplifier: a hybrid series converter with slow-envelope technique and a multilevel converter based on a multiphase buck converter with the minimum time control. In the first architecture, a hybrid topology is composed of a switched buck converter and a linear regulator in series that work together to adjust the output voltage to track the envelope with accuracy. A slow envelope generation algorithm yields a waveform with limited slew rate that is lower than the maximum slew rate of the original envelope. The buck converter's output follows this waveform instead of the original envelope using lower switching frequency, because the waveform has not only reduced slew rate but also reduced bandwidth. In this way, the linear regulator used to filter the waveform has additional power loss. Depending on how much reduction of the slew rate of envelope in order to obtain that waveform, there is a trade-off between the power loss of buck converter related to the switching frequency and the power loss of linear regulator. The optimal point referring to the lowest total power loss of this envelope amplifier is identified with the help of a precise power loss model that is a combination of behavioral and analytic loss model. In addition, the output filter's effect on the response is analyzed. An extra parallel damping filter is needed to eliminate the resonant oscillation of output filter L and C, because the buck converter operates in open loop. The second architecture is a multilevel voltage tracking envelope amplifier. Unlike the converters using multi-sources, a multiphase buck converter is employed to generate the multilevel voltage. In the steady state, the buck converter operates at complete ripple cancellation points of duty cycle. The number of the voltage levels is equal to the number of phases according the characteristics of interleaved buck converter. In the transition, a minimum time control (MTC) for multiphase converter is originally proposed and developed for changing the output voltage of buck converter between different levels. As opposed to conventional minimum time control for multiphase converter with equivalent inductance, the proposed MTC considers the current ripple of each phase based on the fixed phase shift resulting in different control schemes among the phases. The advantage of this control is that all the phase current return to the steady state after the transition so that the next transition can be triggered very soon, which is very favorable for the application of multilevel voltage tracking. Besides, the control is independent on the load condition and not affected by the unbalance of phase current. Like the first architecture, there is also a linear stage with the same function, connected in series with the multiphase buck converter. Since both steady state and transition state of the converter are not strongly related to the switching frequency, it can be reduced for wide bandwidth envelope which is the main consideration of this architecture. The optimization of the second architecture for wider bandwidth envelope is presented including the output filter design, switching frequency and the number of phases. The filter design area is restrained by fast transition and the minimum pulse of hardware. The fast transition needs small filter but the minimum pulse of hardware limitation pushes the filter in opposite way. The converter switching frequency mainly affects the minimum pulse limitation and the power loss. With lower switching frequency, the pulse width in the transition is smaller. The number of phases related to specific application can be optimized in terms of overall efficiency. Another aspect of optimization is improving control strategy. Transition shift allows tracking some parts of envelope that are faster than the hardware can support at the price of complexity. The new transition synchronization method increases the frequency of transition, allowing the multilevel voltage to be closer to the envelope. Both control strategies push the converter to track wider bandwidth envelope than the limitation of power stage. The power loss model of envelope amplifier is detailed and validated by measurements. The power loss mechanism of buck converter has to include the transitions in real time operation, which is different from classical power loss model of synchronous buck converter. This model estimates the system efficiency and play a very important role in optimization process. Finally, the second envelope amplifier architecture is integrated with a Class F amplifier. EER system measurement proves the power saving with the proposed envelope amplifier without disrupting the linearity performance.

Major histocompatibility complex (MHC) class I KbDb −/− deficient mice possess functional CD8+ T cells and natural killer cells

We obtained mice deficient for major histocompatibility complex (MHC) molecules encoded by the H-2K and H-2D genes. H-2 KbDb −/− mice express no detectable classical MHC class I-region associated (Ia) heavy chains, although β2-microglobulin and the nonclassical class Ib proteins examined are expressed normally. KbDb −/− mice have greatly reduced numbers of mature CD8+ T cells, indicating that selection of the vast majority (>90%) of CD8+ T cells cannot be compensated for by β2-microglobulin-associated molecules other than classical H-2K and D locus products. In accord with the greatly reduced number of CD8+ T cells, spleen cells from KbDb −/− mice do not generate cytotoxic responses in primary mixed-lymphocyte cultures against MHC-disparate (allogeneic) cells. However, in vivo priming of KbDb −/− mice with allogeneic cells resulted in strong CD8+ MHC class Ia-specific allogeneic responses. Thus, a minor population of functionally competent peripheral CD8+ T cells capable of strong cytotoxic activity arises in the complete absence of classical MHC class Ia molecules. KbDb −/− animals also have natural killer cells that retain their cytotoxic potential.

Major histocompatibility complex class I-restricted T cells are required for all but the end stages of diabetes development in nonobese diabetic mice and use a prevalent T cell receptor α chain gene rearrangement

Nonobese diabetic (NOD) mice develop insulin-dependent diabetes mellitus due to autoimmune T lymphocyte-mediated destruction of pancreatic β cells. Although both major histocompatibility complex class I-restricted CD8+ and class II-restricted CD4+ T cell subsets are required, the specific role each subset plays in the pathogenic process is still unclear. Here we show that class I-dependent T cells are required for all but the terminal stages of autoimmune diabetes development. To characterize the diabetogenic CD8+ T cells responsible, we isolated and propagated in vitro CD8+ T cells from the earliest insulitic lesions of NOD mice. They were cytotoxic to NOD islet cells, restricted to H-2Kd, and showed a diverse T cell receptor β chain repertoire. In contrast, their α chain repertoire was more restricted, with a recurrent amino acid sequence motif in the complementarity-determining region 3 loop and a prevalence of Vα17 family members frequently joined to the Jα42 gene segment. These results suggest that a number of the CD8+ T cells participating in the initial phase of autoimmune β cell destruction recognize a common structural component of Kd/peptide complexes on pancreatic β cells, possibly a single peptide.

Specific T cell recognition of kinetic isomers in the binding of peptide to class II major histocompatibility complex

Helper T cells are triggered by molecular complexes of antigenic peptides and class II proteins of the major histocompatibility complex . The formation of stable complexes between class II major histocompatibility complex proteins and antigenic peptides is often accompanied by the formation of a short-lived complex. In this report, we describe T cell recognition of two distinct complexes, one short-lived and the other long-lived, formed during the binding of an altered myelin basic protein peptide to I-Ak. One myelin basic protein-specific T cell clone is triggered by only the short-lived complex, and another is triggered by only the stable complex. Thus, a single peptide bound to a particular class II molecule can activate different T cells depending on the conditions of the binding reaction.

A new class of obesity genes encodes leukocyte adhesion receptors

Obesity is a complex disease, and multiple genes contribute to the trait. The description of five genes (ob, db, tub, Ay, and fat) responsible for distinct syndromes of spontaneous monogenic obesity in mice has advanced our knowledge of the genetics of obesity. However, many other genes involved in the expression of this disease remain to be determined. We report here the identification of an additional class of genes involved in the regulation of adipose tissue mass. These genes encode receptors mediating leukocyte adhesion. Mice deficient in intercellular adhesion molecule-1 became spontaneously obese in old age on normal mouse chow or at a young age when provided with a diet rich in fat. Mice deficient in the counterreceptor for intercellular adhesion molecule-1, the leukocyte integrin αMβ2 (Mac-1), showed a similar obesity phenotype. Since all mice consumed approximately the same amount of food as controls, the leukocyte function appears to be in regulating lipid metabolism and/or energy expenditure. Our results indicate that (i) leukocytes play a role in preventing excess body fat deposition and (ii) defects in leukocyte adhesion receptors can result in obesity.

Identification of a new class of protein kinases represented by eukaryotic elongation factor-2 kinase

The several hundred members of the eukaryotic protein kinase superfamily characterized to date share a similar catalytic domain structure, consisting of 12 conserved subdomains. Here we report the existence and wide occurrence in eukaryotes of a protein kinase with a completely different structure. We cloned and sequenced the human, mouse, rat, and Caenorhabditis elegans eukaryotic elongation factor-2 kinase (eEF-2 kinase) and found that with the exception of the ATP-binding site, they do not contain any sequence motifs characteristic of the eukaryotic protein kinase superfamily. Comparison of different eEF-2 kinase sequences reveals a highly conserved region of ≈200 amino acids which was found to be homologous to the catalytic domain of the recently described myosin heavy chain kinase A (MHCK A) from Dictyostelium. This suggests that eEF-2 kinase and MHCK A are members of a new class of protein kinases with a novel catalytic domain structure.

Role of the major histocompatibility complex class II Ea gene in lupus susceptibility in mice

The gene(s) encoded within major histocompatibility complex (MHC) act as one of the major genetic elements contributing to the susceptibility of murine systemic lupus erythematosus (SLE). We have recently demonstrated that lupus susceptibility is more closely linked to the I-E− H-2b haplotype than to the I-E+ H-2d haplotype in lupus-prone BXSB and (NZB × BXSB)F1 hybrid mice. To investigate whether the reduced susceptibility to SLE in H-2d mice is related to the expression of the MHC class II Ea gene (absent in H-2b mice), we determined the possible role of the Ea gene as a lupus protective gene in mice. Our results showed that (i) the development of SLE was almost completely prevented in BXSB (H-2b) mice expressing two copies of the Ead transgene at the homozygous level as well as in BXSB H-2k (I-E+) congenic mice as for H-2d BXSB mice, and (ii) the expression of two functional Ea (transgenic and endogenous) genes in either H-2d/b (NZB × BXSB)F1 or H-2k/b (MRL × BXSB)F1 mice provided protection from SLE at levels comparable to those conferred by the H-2d/d or H-2k/k haplotype. In addition, the level of the Ea gene-mediated protection appeared to be dependent on the genetic susceptibility to SLE in individual lupus-prone mice. Our results indicate that the reduced susceptibility associated with the I-E+ H-2d and H-2k haplotypes (versus the I-E− H-2b haplotype) is largely, if not all, contributed by the apparent autoimmune suppressive effect of the Ea gene, independently of the expression of the I-A or other MHC-linked genes.

Role of the Box C/D Motif in Localization of Small Nucleolar RNAs to Coiled Bodies and Nucleoli

Small nucleolar RNAs (snoRNAs) are a large family of eukaryotic RNAs that function within the nucleolus in the biogenesis of ribosomes. One major class of snoRNAs is the box C/D snoRNAs named for their conserved box C and box D sequence elements. We have investigated the involvement of cis-acting sequences and intranuclear structures in the localization of box C/D snoRNAs to the nucleolus by assaying the intranuclear distribution of fluorescently labeled U3, U8, and U14 snoRNAs injected into Xenopus oocyte nuclei. Analysis of an extensive panel of U3 RNA variants showed that the box C/D motif, comprised of box C′, box D, and the 3′ terminal stem of U3, is necessary and sufficient for the nucleolar localization of U3 snoRNA. Disruption of the elements of the box C/D motif of U8 and U14 snoRNAs also prevented nucleolar localization, indicating that all box C/D snoRNAs use a common nucleolar-targeting mechanism. Finally, we found that wild-type box C/D snoRNAs transiently associate with coiled bodies before they localize to nucleoli and that variant RNAs that lack an intact box C/D motif are detained within coiled bodies. These results suggest that coiled bodies play a role in the biogenesis and/or intranuclear transport of box C/D snoRNAs.

Class VI Unconventional Myosin is Required for Spermatogenesis in Drosophila

We have identified partial loss of function mutations in class VI unconventional myosin, 95F myosin, which results in male sterility. During spermatogenesis the germ line precursor cells undergo mitosis and meiosis to form a bundle of 64 spermatids. The spermatids remain interconnected by cytoplasmic bridges until individualization. The process of individualization involves the formation of a complex of cytoskeletal proteins and membrane, the individualization complex (IC), around the spermatid nuclei. This complex traverses the length of each spermatid resolving the shared membrane into a single membrane enclosing each spermatid. We have determined that 95F myosin is a component of the IC whose function is essential for individualization. In wild-type testes, 95F myosin localizes to the leading edge of the IC. Two independent mutations in 95F myosin reduce the amount of 95F myosin in only a subset of tissues, including the testes. This reduction of 95F myosin causes male sterility as a result of defects in spermatid individualization. Germ line transformation with the 95F myosin heavy chain cDNA rescues the male sterility phenotype. IC movement is aberrant in these 95F myosin mutants, indicating a critical role for 95F myosin in IC movement. This report is the first identification of a component of the IC other than actin. We propose that 95F myosin is a motor that participates in membrane reorganization during individualization.

Immune response in human melanoma after transfer of an allogeneic class I major histocompatibility complex gene with DNA–liposome complexes

Analysis of the antitumor immune response after gene transfer of a foreign major histocompatibility complex class I protein, HLA-B7, was performed. Ten HLA-B7-negative patients with stage IV melanoma were treated in an effort to stimulate local tumor immunity. Plasmid DNA was detected within treated tumor nodules, and RNA encoding recombinant HLA-B7 or HLA-B7 protein was demonstrated in 9 of 10 patients. T cell migration into treated lesions was observed and tumor-infiltrating lymphocyte reactivity was enhanced in six of seven and two of two patients analyzed, respectively. In contrast, the frequency of cytotoxic T lymphocyte against autologous tumor in circulating peripheral blood lymphocytes was not altered significantly, suggesting that peripheral blood lymphocyte reactivity is not indicative of local tumor responsiveness. Local inhibition of tumor growth was detected after gene transfer in two patients, one of whom showed a partial remission. This patient subsequently received treatment with tumor-infiltrating lymphocytes derived from gene-modified tumor, with a complete regression of residual disease. Thus, gene transfer with DNA–liposome complexes encoding an allogeneic major histocompatibility complex protein stimulated local antitumor immune responses that facilitated the generation of effector cells for immunotherapy of cancer.

The Export of Major Histocompatibility Complex Class I Molecules from the Endoplasmic Reticulum of Rat Brown Adipose Cells Is Acutely Stimulated by Insulin

Major histocompatibility complex class I (MHC-I) molecules have been implicated in several nonimmunological functions including the regulation and intracellular trafficking of the insulin-responsive glucose transporter GLUT4. We have used confocal microscopy to compare the effects of insulin on the intracellular trafficking of MHC-I and GLUT4 in freshly isolated rat brown adipose cells. We also used a recombinant vaccinia virus (rVV) to express influenza virus hemagglutinin (HA) as a generic integral membrane glycoprotein to distinguish global versus specific enhancement of protein export from the endoplasmic reticulum (ER) in response to insulin. In the absence of insulin, MHC-I molecules largely colocalize with the ER-resident protein calnexin and remain distinct from intracellular pools of GLUT4. Surprisingly, insulin induces the rapid export of MHC-I molecules from the ER with a concomitant approximately three-fold increase in their level on the cell surface. This ER export is blocked by brefeldin A and wortmannin but is unaffected by cytochalasin D, indicating that insulin stimulates the rapid transport of MHC-I molecules from the ER to the plasma membrane via the Golgi complex in a phosphatidyl-inositol 3-kinase–dependent and actin-independent manner. We further show that the effect of insulin on MHC-I molecules is selective, because insulin does not affect the intracellular distribution or cell-surface localization of rVV-expressed HA. These results demonstrate that in rat brown adipose cells MHC-I molecule export from the ER is stimulated by insulin and provide the first evidence that the trafficking of MHC-I molecules is acutely regulated by a hormone.

Distinct Cyclin D Genes Show Mitotic Accumulation or Constant Levels of Transcripts in Tobacco Bright Yellow-2 Cells1

The commitment of eukaryotic cells to division normally occurs during the G1 phase of the cell cycle. In mammals D-type cyclins regulate the progression of cells through G1 and therefore are important for both proliferative and developmental controls. Plant CycDs (D-type cyclin homologs) have been identified, but their precise function during the plant cell cycle is unknown. We have isolated three tobacco (Nicotiana tabacum) CycD cyclin cDNAs: two belong to the CycD3 class (Nicta;CycD3;1 and Nicta;CycD3;2) and the third to the CycD2 class (Nicta;CycD2;1). To uncouple their cell-cycle regulation from developmental control, we have used the highly synchronizable tobacco cultivar Bright Yellow-2 in a cell-suspension culture to characterize changes in CycD transcript levels during the cell cycle. In cells re-entering the cell cycle from stationary phase, CycD3;2 was induced in G1 but subsequently remained at a constant level in synchronous cells. This expression pattern is consistent with a role for CycD3;2, similar to mammalian D-type cyclins. In contrast, CycD2;1 and CycD3;1 transcripts accumulated during mitosis in synchronous cells, a pattern of expression not normally associated with D-type cyclins. This could suggest a novel role for plant D-type cyclins during mitosis.

Oncolytic herpes simplex virus vector with enhanced MHC class I presentation and tumor cell killing

Oncolytic herpes simplex virus type 1 (HSV-1) vectors are promising therapeutic agents for cancer. Their efficacy depends on the extent of both intratumoral viral replication and induction of a host antitumor immune response. To enhance these properties while employing ample safeguards, two conditionally replicating HSV-1 vectors, termed G47Δ and R47Δ, have been constructed by deleting the α47 gene and the promoter region of US11 from γ34.5-deficient HSV-1 vectors, G207 and R3616, respectively. Because the α47 gene product is responsible for inhibiting the transporter associated with antigen presentation (TAP), its absence led to increased MHC class I expression in infected human cells. Moreover, some G47Δ-infected human melanoma cells exhibited enhanced stimulation of matched antitumor T cell activity. The deletion also places the late US11 gene under control of the immediate-early α47 promoter, which suppresses the reduced growth properties of γ34.5-deficient mutants. G47Δ and R47Δ showed enhanced viral growth in a variety of cell lines, leading to higher virus yields and enhanced cytopathic effect in tumor cells. G47Δ was significantly more efficacious in vivo than its parent G207 at inhibiting tumor growth in both immune-competent and immune-deficient animal models. Yet, when inoculated into the brains of HSV-1-sensitive A/J mice at 2 × 106 plaque forming units, G47Δ was as safe as G207. These results suggest that G47Δ may have enhanced antitumor activity in humans.

N-Acylethanolamines: Formation and Molecular Composition of a New Class of Plant Lipids1

Recently, the biosynthesis of an unusual membrane phospholipid, N-acylphosphatidylethanolamine (NAPE), was found to increase in elicitor-treated tobacco (Nicotiana tabacum L.) cells (K.D. Chapman, A. Conyers-Hackson, R.A. Moreau, S. Tripathy [1995] Physiol Plant 95: 120–126). Here we report that before induction of NAPE biosynthesis, N-acylethanolamine (NAE) is released from NAPE in cultured tobacco cells 10 min after treatment with the fungal elicitor xylanase. In radiolabeling experiments [14C]NAE (labeled on the ethanolamine carbons) increased approximately 6-fold in the culture medium, whereas [14C]NAPE associated with cells decreased approximately 5-fold. Two predominant NAE molecular species, N-lauroylethanolamine and N-myristoylethanolamine, were specifically identified by gas chromatography-mass spectrometry in lipids extracted from culture medium, and both increased in concentration after elicitor treatment. NAEs were found to accumulate extracellularly only. A microsomal phospholipase D activity was discovered that formed NAE from NAPE; its activity in vitro was stimulated about 20-fold by mastoparan, suggesting that NAPE hydrolysis is highly regulated, perhaps by G-proteins. Furthermore, an NAE amidohydrolase activity that catalyzed the hydrolysis of NAE in vitro was detected in homogenates of tobacco cells. Collectively, these results characterize structurally a new class of plant lipids and identify the enzymatic machinery involved in its formation and inactivation in elicitor-treated tobacco cells. Recent evidence indicating a signaling role for NAPE metabolism in mammalian cells (H.H.O. Schmid, P.C. Schmid, V. Natarajan [1996] Chem Phys Lipids 80: 133–142) raises the possibility that a similar mechanism may operate in plant cells.