NADH-Glutamate Synthase in Alfalfa Root Nodules. Immunocytochemical Localization1

In root nodules of alfalfa (Medicago sativa L.), N2 is reduced to NH4+ in the bacteroid by the nitrogenase enzyme and then released into the plant cytosol. The NH4+ is then assimilated by the combined action of glutamine synthetase (EC 6.3.1.2) and NADH-dependent Glu synthase (NADH-GOGAT; EC 1.4.1.14) into glutamine and Glu. The alfalfa nodule NADH-GOGAT protein has a 101-amino acid presequence, but the subcellular location of the protein is unknown. Using immunocytochemical localization, we determined first that the NADH-GOGAT protein is found throughout the infected cell region of both 19- and 33-d-old nodules. Second, in alfalfa root nodules NADH-GOGAT is localized predominantly to the amyloplast of infected cells. This finding, together with earlier localization and fractionation studies, indicates that in alfalfa the infected cells are the main location for the initial assimilation of fixed N2.

Glycolytic Flux Is Adjusted to Nitrogenase Activity in Nodules of Detopped and Argon-Treated Alfalfa Plants1

To investigate the short-term (30–240 min) interactions among nitrogenase activity, NH4+ assimilation, and plant glycolysis, we measured the concentrations of selected C and N metabolites in alfalfa (Medicago sativa L.) root nodules after detopping and during continuous exposure of the nodulated roots to Ar:O2 (80:20, v/v). Both treatments caused an increase in the ratios of glucose-6-phosphate to fructose-1,6-bisphosphate, fructose-6-phosphate to fructose-1,6-bisphosphate, phosphoenolpyruvate (PEP) to pyruvate, and PEP to malate. This suggested that glycolytic flux was inhibited at the steps catalyzed by phosphofructokinase, pyruvate kinase, and PEP carboxylase. In the Ar:O2-treated plants the apparent inhibition of glycolytic flux was reversible, whereas in the detopped plants it was not. In both groups of plants the apparent inhibition of glycolytic flux was delayed relative to the decline in nitrogenase activity. The decline in nitrogenase activity was followed by a dramatic increase in the nodular glutamate to glutamine ratio. In the detopped plants this was coincident with the apparent inhibition of glycolytic flux, whereas in the Ar:O2-treated plants it preceded the apparent inhibition of glycolytic flux. We propose that the increase in the nodular glutamate to glutamine ratio, which occurs as a result of the decline in nitrogenase activity, may act as a signal to decrease plant glycolytic flux in legume root nodules.

Purification of the Trehalase GMTRE1 from Soybean Nodules and Cloning of Its cDNA. GMTRE1 Is Expressed at a Low Level in Multiple Tissues1

Trehalose (α-d-glucopyranosyl-1,1-α-d-glucopyranoside), a disaccharide widespread among microbes and lower invertebrates, is generally believed to be nonexistent in higher plants. However, the recent discovery of Arabidopsis genes whose products are involved in trehalose synthesis has renewed interest in the possibility of a function of trehalose in higher plants. We previously showed that trehalase, the enzyme that degrades trehalose, is present in nodules of soybean (Glycine max [L.] Merr.), and we characterized the enzyme as an apoplastic glycoprotein. Here we describe the purification of this trehalase to homogeneity and the cloning of a full-length cDNA encoding this enzyme, named GMTRE1 (G. max trehalase 1). The amino acid sequence derived from the open reading frame of GMTRE1 shows strong homology to known trehalases from bacteria, fungi, and animals. GMTRE1 is a single-copy gene and is expressed at a low but constant level in many tissues.

Song selectivity and sensorimotor signals in vocal learning and production

Bird song, like human speech, is a learned vocal behavior that requires auditory feedback. Both as juveniles, while they learn to sing, and as adults, songbirds use auditory feedback to compare their own vocalizations with an internal model of a target song. Here we describe experiments that explore a role for the songbird anterior forebrain pathway (AFP), a basal ganglia-forebrain circuit, in evaluating song feedback and modifying vocal output. First, neural recordings in anesthetized, juvenile birds show that single AFP neurons are specialized to process the song stimuli that are compared during sensorimotor learning. AFP neurons are tuned to both the bird's own song and the tutor song, even when these stimuli are manipulated to be very different from each other. Second, behavioral experiments in adult birds demonstrate that lesions to the AFP block the deterioration of song that normally follows deafening. This observation suggests that deafening results in an instructive signal, indicating a mismatch between feedback and the internal song model, and that the AFP is involved in generating or transmitting this instructive signal. Finally, neural recordings from behaving birds reveal robust singing-related activity in the AFP. This activity is likely to originate from premotor areas and could be modulated by auditory feedback of the bird's own voice. One possibility is that this activity represents an efference copy, predicting the sensory consequences of motor commands. Overall, these studies illustrate that sensory and motor processes are highly interrelated in this circuit devoted to vocal learning, as is true for brain areas involved in speech.

On cortical coding of vocal communication sounds in primates

Understanding how the brain processes vocal communication sounds is one of the most challenging problems in neuroscience. Our understanding of how the cortex accomplishes this unique task should greatly facilitate our understanding of cortical mechanisms in general. Perception of species-specific communication sounds is an important aspect of the auditory behavior of many animal species and is crucial for their social interactions, reproductive success, and survival. The principles of neural representations of these behaviorally important sounds in the cerebral cortex have direct implications for the neural mechanisms underlying human speech perception. Our progress in this area has been relatively slow, compared with our understanding of other auditory functions such as echolocation and sound localization. This article discusses previous and current studies in this field, with emphasis on nonhuman primates, and proposes a conceptual platform to further our exploration of this frontier. It is argued that the prerequisite condition for understanding cortical mechanisms underlying communication sound perception and production is an appropriate animal model. Three issues are central to this work: (i) neural encoding of statistical structure of communication sounds, (ii) the role of behavioral relevance in shaping cortical representations, and (iii) sensory–motor interactions between vocal production and perception systems.

Uridine 5′-Diphosphate-Glucose Dehydrogenase from Soybean Nodules

A highly purified preparation of uridine 5′-diphosphate (UDP)-glucose (Glc) dehydrogenase (DH; EC 1.1.1.22) has been characterized from soybean (Glycine max L.) nodules. The enzyme had native and subunit molecular masses of approximately 272 and 50 kD, respectively. UDP-Glc DH displayed typical hyperbolic substrate kinetics and had Km values for UDP-Glc and NAD+ of 0.05 and 0.12 mm, respectively. Thymidine 5′-diphosphate-Glc and UDP-galactose could replace UDP-Glc as the sugar nucleotide substrate to some extent, but the enzyme had no activity with NADP+. Soybean nodule UDP-Glc DH was labile in the absence of NAD+ and was inhibited by a heat-stable, low-molecular-mass solute in crude extracts of soybean nodules. UDP-Glc DH was also isolated from developing soybean seeds and shoots of 5-d-old wheat and canola seedlings and was shown to have similar affinities for UDP-Glc and NAD+ as those of the soybean nodule enzyme. UDP-Glc DH from all of these sources was most active in young, rapidly growing tissues.

Symbiotic induction of a MADS-box gene during development of alfalfa root nodules.

In response to infection by Rhizobium, highly differentiated organs called nodules form on legume roots. Within these organs, the symbiotic association between the host plant and bacteria is established. A putative plant transcription factor, NMH7, has been identified in alfalfa root nodules. nmh7 contains a MADS-box DNA-binding region and shows homology to flower homeotic genes. This gene is a member of a multigene family in alfalfa and was identified on the basis of nucleic acid homology to plant regulatory protein genes (MADS-box-containing genes) from Antirrhinum and Arabidopsis. RNA analysis and in situ hybridization showed that expression of this class of regulatory genes is limited to the infected cells of alfalfa root nodules and is likely to be involved in the signal transduction pathway initiated by the bacterial symbiont, Rhizobium meliloti. The expression of nmh7 in a root-derived organ is unusual for this class of regulatory genes.

Relações entre comportamento vocal, imunidade e níveis plasmáticos de corticosterona e testosterona em sapos (Rhinella granulosa)

A exibição do comportamento vocal em muitas espécies de anfíbios anuros é relacionada aos níveis de hormônios esteroides gonadais e interrenais. Esses hormônios poderiam mediar a relação entre intensidade de sinais e imunidade, pois estão envolvidos no desenvolvimento das características sexuais secundárias, comportamento de corte e mobilização de reservas energéticas durante a atividade reprodutiva, enquanto apresentam também efeitos imunomoduladores. Nesse sentido, o objetivo deste trabalho foi explorar as relações entre comportamento reprodutivo, imunidade e níveis plasmáticos de testosterona e corticosterona em machos do sapo do semiárido brasileiro, Rhinella granulosa, em atividade reprodutiva e após manipulação hormonal. A precipitação foi o principal determinante ambiental para o aumento dos níveis de testosterona e corticosterona circulantes em machos de R. granulosa, estimulando o comportamento de vocalização. As relações fisiológicas encontradas indicam que os altos níveis plasmáticos de testosterona nos primeiros dias após a chuva devem promover o início do turno vocal, porém a corticosterona deve modular o esforço de vocalização. De forma geral, a exacerbação do comportamento vocal de R. granulosa tem efeitos negativos sobre a imunocompetência, porém alguns indivíduos que apresentam maiores concentrações plasmáticas de corticosterona apresentam concomitantemente alto esforço vocal e alta imunidade. De acordo, a aplicação transdérmica de corticosterona promoveu elevação aguda dos níveis plasmáticos deste glicocorticoide, bem como um aumento da função imune. Assim, apesar de a atenção principal ser comumente colocada no papel da testosterona na mediação de sinais honestos, nossos resultados corroboram a importância da corticosterona na mediação da expressão do comportamento de corte e imunocompetência em machos R. granulosa

Aplicação de uma rede neural artificial para a avaliação da rugosidade e soprosidade vocal

A avaliação perceptivo-auditiva tem papel fundamental no estudo e na avaliação da voz, no entanto, por ser subjetiva está sujeita a imprecisões e variações. Por outro lado, a análise acústica permite a reprodutibilidade de resultados, porém precisa ser aprimorada, pois não analisa com precisão vozes com disfonias mais intensas e com ondas caóticas. Assim, elaborar medidas que proporcionem conhecimentos confiáveis em relação à função vocal resulta de uma necessidade antiga dentro desta linha de pesquisa e atuação clínica. Neste contexto, o uso da inteligência artificial, como as redes neurais artificiais, indica ser uma abordagem promissora. Objetivo: Validar um sistema automático utilizando redes neurais artificiais para a avaliação de vozes rugosas e soprosas. Materiais e métodos: Foram selecionadas 150 vozes, desde neutras até com presença em grau intenso de rugosidade e/ou soprosidade, do banco de dados da Clínica de Fonoaudiologia da Faculdade de Odontologia de Bauru (FOB/USP). Dessas vozes, 23 foram excluídas por não responderem aos critérios de inclusão na amostra, assim utilizaram-se 123 vozes. Procedimentos: avaliação perceptivo-auditiva pela escala visual analógica de 100 mm e pela escala numérica de quatro pontos; extração de características do sinal de voz por meio da Transformada Wavelet Packet e dos parâmetros acústicos: jitter, shimmer, amplitude da derivada e amplitude do pitch; e validação do classificador por meio da parametrização, treino, teste e avaliação das redes neurais artificiais. Resultados: Na avaliação perceptivo-auditiva encontrou-se, por meio do teste Coeficiente de Correlação Intraclasse (CCI), concordâncias inter e intrajuiz excelentes, com p = 0,85 na concordância interjuízes e p variando de 0,87 a 0,93 nas concordâncias intrajuiz. Em relação ao desempenho da rede neural artificial, na discriminação da soprosidade e da rugosidade e dos seus respectivos graus, encontrou-se o melhor desempenho para a soprosidade no subconjunto composto pelo jitter, amplitude do pitch e frequência fundamental, no qual obteve-se taxa de acerto de 74%, concordância excelente com a avaliação perceptivo-auditiva da escala visual analógica (0,80 no CCI) e erro médio de 9 mm. Para a rugosidade, o melhor subconjunto foi composto pela Transformada Wavelet Packet com 1 nível de decomposição, jitter, shimmer, amplitude do pitch e frequência fundamental, no qual obteve-se 73% de acerto, concordância excelente (0,84 no CCI), e erro médio de 10 mm. Conclusão: O uso da inteligência artificial baseado em redes neurais artificiais na identificação, e graduação da rugosidade e da soprosidade, apresentou confiabilidade excelente (CCI > 0,80), com resultados semelhantes a concordância interjuízes. Dessa forma, a rede neural artificial revela-se como uma metodologia promissora de avaliação vocal, tendo sua maior vantagem a objetividade na avaliação.

Heavy metal contents in Fe-Mn nodules and crusts from R/V Vityaz polygons and stations in the East Indian Ocean

Regional variations in abundance, morphology, and chemical composition of Fe-Mn nodules have a zonal character. Due to circumcontinental zonality of terrigenous sedimentation the main mass of the nodules occurs in the pelagic part of the ocean, in areas of minimal sedimentation rates. In spatial variations in morphology and chemical composition of the nodules the latitudinal zonality is very clear and associated with latitudinal changes in facial conditions of sedimentation. Elevated contents of Mn, Ni, and Cu and of Mn/Fe ratio occur in nodules from the radiolarian belt. Changes of chemical composition of the nodules with depth (vertical zonality of mineralization) are confirmed. Local variations in abundance, morphology and chemical composition of the nodules are caused by ruggedness of relief and depth variations, variations in sedimentation rate, age of ore formation, intensity of diagenetic redistribution of metals.

(Table 1, page 1157) DOWNWIND expedition bottom photo stations showing abundant manganese nodules

Manganese nodules were investigated during the Downwind Expedition, a part of the International Geophysical Year programme of the Scripps Institution of Oceanography of the University of California. Attempts were made to collect bottom photographs, cores and dredge hauls in the same areas, to measure the distribution at the surface and in depth, and to obtain large samples for physical and chemical analysis.

(Table 1, page 24), Geochemical cmposition of nodules and crust from the Indain Ocean recovered from R/V Marion Dufresne expeditions

Mineralogical analysis of manganese nodules and crusts collected from Indian ocean aboard Marion Dufresne points to a depth and regional control upon the manganese oxide association: vernadite - birnessite and vernadite - todorokite. Moreover, progressive changes in the vernadite/birnessite ratio as a function of time is clearly seen. Magnetite and titano-magnetite in quantities similar to those of framboidal pyrite in manganese nodules are outlined for the first time. Study of the distribution of metals (Mn, Fe, Ni, Cu, Co) shows a strong latitudinal and regional dependence that may be connected to high productivity zones and to bottom water properties. The problem of mineralogical control on the chemical composition is approached. Finally, it results that any interpretations taking into account all these data haveway to give to the variability of sea-water properties (pH, oxygenation, motions) the prominent control upon manganese nodules composition.

(Table 2) Chemical composition of upper and lower surfaces of Fe-Mn nodules from the Eastern Subequatorial Pacific

Materials of polygon exploration during Cruise 41 of R/V Dmitry Mendeleev showed that diagenetic and sedimentary-diagenetic nodules close in morphology, texture, and composition vary greatly in size and productivity. Local variations in productivity of this nodule type in pelagic areas of the Pacific Ocean are closely connected with thickness of underlying clayey-radiolarian oozes.

(Table 8.4, Page 220-223) Chemical composition of different growth structures of manganese nodules, SONNE cruise SO78

In the nodule field of the Peru Basin, situated south of the zone of high bioproductivity, a relatively high flux of biogenic matter explains a distinct redox boundary at about 10 cm depth separating very soft oxic surface sediments from stiffer suboxic sediments. Maximum abundance (50 kg/m**2) of diagenetic nodules is found near the calcite compensation depth (CCD), currently at 4250 m. There, the accretion rate of nodules is much higher (100 mm/Ma) than on ridges (5 mm/Ma). Highest accretion rates are found at the bottom of large nodules that repeatedly sink to a level immediately above the redox boundary. There, distinct diagenetic growth conditions prevail and layers of dense laminated Mn oxide of very pure todorokite are formed. The layering of nodules is mainly the result of organisms moving nodules within the oxic surface sediment from diagenetic to hydrogenetic environments. The frequency of such movements is much higher than that of climatic changes. Two types of nodule burial occur in the Peru Basin. Large nodules are less easily moved by organisms and become buried. Consequently, buried nodules generally are larger than surface nodules. This type of burial predominates in basins. At ridges where smaller nodules prevail, burial is mainly controlled by statistical selection where some nodules are not moved up by organisms.