The specificity of angiotensin II receptor binding in rat brain

125I-angiotensin II (125I-AII) binding was examined in the hypothalamic-thalamic-septal-midbrain (HTSM) region of HLA-Wistar rats in the presence of CNS-active agents. Angiotensin I, II, and III and saralasin competed for 125 I-AII binding, whereas structurally unrelated peptides such as arginine and lysine vasopressin, oxytocin, LHRH, TRH, bradykinin, and substance P did not. In contrast, ACTH and neurotensin exhibited a weak, dose-dependent competition for 125 I-AII binding. The relative potencies of AII, AI, neurotensin and ACTH were 100:1:0.1:0.05, respectively. Neurotensin and ACTH competition was not additive with AII suggesting interaction at shared binding sites. Most importantly, a wide variety of other CNS active agents such as methyldopa, naloxone, catecholamines, clondidine, and reserpine, failed to inhibit 125 I-AII binding, thus further defining the specificity of the CNS AII receptor.     

Population Dynamics of Spotted Owls in the Sierra Nevada,California

ABSTRACT The California spotted owl (Strix occidentalis occidentalis) is the only spotted owl subspecies not listed as threatened or endangered under the United States Endangered Species Act despite petitions to list it as threatened. We conducted a meta-analysis of population data for 4 populations in the southem Cascades and Sierra Nevada, California, USA, from 1990 to 2005 to assist a listing evaluation by the United States Fish and Wildlife Service. Our study areas (from N to S) were on the Lassen National Forest (LAS), Eldorado National Forest (ELD), Sierra National Forest (SIE), and Sequoia and Kings Canyon National Parks (SKC). These study areas represented a broad spectrum of habitat and management conditions in these mountain ranges. We estimated apparent survival probability, reproductive output, and rate of population change for spotted owls on individual study areas and for all study areas combined (meta-analysis) using model selection or model-averaging based on maximum-likelihood estimation. We followed a formal protocol to conduct this analysis that was similar to other spotted owl meta-analyses. Consistency of field and analytical methods among our studies reduced confounding methodological effects when evaluating results. We used 991 marked spotted owls in the analysis of apparent survival. Apparent survival probability was higher for adult than for subadult owls. There was little difference in apparent survival between male and female owls. Model-averaged mean estimates of apparent survival probability of adult owls varied from 0.811 ± 0.021 for females at LAS to 0.890 ± 0.016 for males at SKC. Apparent survival increased over time for owls of all age classes at LAS and SIE, for adults at ELD, and for second-year subadults and adults at SKC. The meta-analysis of apparent survival, which included only adult owls, confirmed an increasing trend in survival over time. Survival rates were higher for owls on SKC than on the other study areas. We analyzed data from 1,865 observations of reproductive outcomes for female spotted owls. The proportion of subadult females among all territorial females of known age ranged from 0.00 to 0.25 among study areas and years. The proportion of subadults among female spotted owls was negatively related to reproductive output (no. of young fledged/territorial F owl) for ELD and SIE. Eldorado study area and LAS showed an alternate-year trend in reproductive output, with higher output in even-numbered years. Mean annual reproductive output was 0.988 ± 0.154 for ELD, 0.624 ± 0.140 for LAS, 0.478 ± 0.106 for SIE, and 0.555 ± 0.110 for SKC. Eldorado Study Area exhibited a declining trend and the greatest variation in reproductive output over time, whereas SIE and SKC, which had the lowest reproductive output, had the lowest temporal variation. Meta-analysis confirmed that reproductive output varied among study areas. Reproductive output was highest for adults, followed by second-year subadults, and then by first-year subadults. We used 842 marked subadult and adult owls to estimate population rate of change. Modeling indicated that Λ _t (Λ _t is the finite rate of population change estimated using the reparameterized Jolly–Seber estimator [Pradel 1996]) was either stationary (LAS and SIE) or increasing after an initial decrease (ELD and SKC). Mean estimated Λ _t for the 4 study areas was 1.007 (95% CI = 0.952–1.066) for ELD; 0.973 (95% CI = 0.946–1.001) for LAS; 0.992 (95% CI = 0.966–1.018) for SIE; and 1.006 (95% CI = 0.947–1.068) for SKC. The best meta-analysis model of population trend indicated that Λ varied across time but was similar in trend among the study areas. Our estimates of realized population change (Δ _t ; Franklin et al. 2004), which we estimated as the product 1 λ₃, were based on estimates of Λ _t from individual study areas and did not require estimating annual population size for each study area. Trends represented the proportion of the population size in the first year that remained in each subsequent year. Similar to λ₄ on which they were based, these λ_k-1 showed evidence of decline over the study period for LAS and SIE. The best model indicated recruitment of male and female adult and subadults varied from 0.10 to 0.31 new territorial individuals at time t/number of territorial individuals at time t–1 and similarly among areas. We also conducted a population viability analysis (PVA) based on results of our meta-analysis. This PVA was of limited utility for ELD and SKC study areas because 95% confidence intervals on the probability of decline or increase spanned the interval [0, 1] within 5–10 years. When we restricted inferences to 7 years, estimated probability of a >10% decline for SIE was 0.41 (95% CI = 0.09–0.78); for LAS the probability was 0.64 (95% CI = 0.27–0.94). In contrast, estimated probability of a >10% increase in 7 years for SIE was 0.23 (95% CI = 0.01–0.55) and for LAS was 0.10 (95% CI = 0.00–0.34). For comparisons, we simulated a PVA for a hypothetical population with mean Λ = 1.0 and the same temporal variation as observed in our owl populations. Our PVA suggested that both the SIE and LAS populations had higher probabilities of declining in a 7-year period than increasing but that it would be difficult to determine if a population was in a slight gradual decline. Our analysis and the repository of information on our 4 study populations provide a data-rich template for managers to monitor impacts of future management actions on the owl. Specifically, our data can be used to evaluate the effect of management strategies on spotted owls that are being implemented by the United States Forest Service to reduce the risk of wildfire in the Sierra Nevada ecosystem. Our information also provides baseline information for evaluating the status of the owl for potential listing as a threatened species by the United States Fish and Wildlife Service. RESUMEN El búho californiano manchado (Strix occidentalis occidentalis) es la única subespecie de búhos manchados que no está listada como amenazada o en peligro de extinción en el Acta de E.E.U.U. para las Especies en Peligro de Extinción a pesar de las peticiones para que sea incluida en la lista como una especie amenazada. Nosotros realizamos un meta-análisis de los datos de la población de 4 poblaciones del sur de Cascades y de la Sierra Nevada, California desde 1990 hasta 2005 como ayuda a una evaluación de listado hecha por el U.S Fish and Wildlife Service. Nuestras áreas de estudio (de norte a sur) estuvieron localizadas en el Bosque Nacional Lassen (LAS), en el Bosque Nacional Eldorado (ELD), en el Bosque Nacional Sierra (SIE) y en los Parques Nacionales Sequoia y Kings Canyon (SKC). Estas áreas de estudio representaron un amplio espectro del hábitat y de las condiciones de manejo en estas cadenas de montañas. Nosotros calculamos la probabilidad de supervivencia aparente, el volumen de reproducción y el cambio en la tasa de población de los búhos manchados en áreas de estudio individuales y para todas las áreas de estudio combinadas (meta-análisis) utilizando selección de modelos o promediando modelos basados en la estimación de máxima probabilidad. Seguimos un protocolo formal para realizar este análisis que fuera similar a otros meta-análisis con búhos manchados. La consistencia del campo y los métodos analíiticos en nuestros estudios redujeron la confusión de efectos metodológicos al evaluar los resultados. Utilizamos 991 búhos manchados marcados en el análisis de supervivencia aparente. La probabilidad de supervivencia aparente fue más alta para búhos adultos que para subadultos. Hubo poca diferencia en la supervivencia aparente entre hembras y machos. Para los modelos promediados, los cálculos de la media de la probabilidad de supervivencia aparente para búhos adultos tuvo una variación de 0.811 ± 0.021 para hembras en LAS a 0.890 ± 0.016 para machos en SKA. La supervivencia aparente aumentó con el tiempo para los búhos de todos los grupos de edad en LAS y SIE, para adultos en ELD, y para subadultos del segundo año y para adultos en SKC. El meta-análisis de supervivencia aparente, que incluyó únicamente a búhos adultos, confirmó una tendencia al aumento en la supervivencia con el tiempo. Las tasas de supervivencia fueron más altas para los búhos en SKC que en las otras áreas de estudio. Analizamos información de 1.865 observaciones de resultados de reproducciones para búhos manchados hembra. La proporción de hembras subadultas entre todas las hembras territoriales de edad conocida fluctuó de 0.00 a 0.25 a través de las áreas de estudio y de los años. La proporción de subadultos entre los búhos manchados hembra estuvo relacionada negativamente con el volumen de reproducción (número de pichones emplumados por búho hembra territorial) para ELD y SIE. ELD y LAS mostraron una tendencia anual alternada en el volumen de reproducción, con un volumen mayor en los años pares. La media del volumen de reproducción anual fue 0.988 ± 0.154 para ELD, 0.624 ± 0.140 para LAS, 0.478 ± 0.106 para SIE y 0.555 ± 0.154 para SKC. ELD exhibió una tendencia a disminuir y la variación más alta en el volumen de reproducción a través del tiempo; mientras que SIE y SKC, que tuvieron el más bajo volumen de reproducción, tuvieron la menor variación temporal. El meta-análisis confirmó que el volumen de reproducción varió entre las áreas de estudio. El volumen de reproducción fue más alto para adultos, seguido por subadultos del segundo año, y luego por subadultos del primer año. Nosotros utilizamos 842 búhos marcados, adultos y subadultos, para calcular el índice de cambio de la población. La selección de modelos indicó que Λ _t era, o relativamente fija (LAS y SIE) o aumentaba después de una disminución inicial (ELD y SKC). La media calculada Λ _t para las cuatro áreas de estudio fue: 1.007 (95% CI = 0.952–1.066) para ELD; 0.973 (95% CI = 0.946–1.001) para LAS; 0.992 (95% CI = 0.966–1.018) para SIE; y 1.006 (95% CI = 0.947–1.068) para SKC. El mejor modelo de meta-análisis de la tendencia de población indicó que Λ variaba con el tiempo pero que era una tendencia similar entre las áreas de estudio. Nuestros cálculos sobre el cambio de población realizado (Δ _t ) se basaron en los cálculos de Λ _t de las áreas de estudio individuales y no requirieron calcular el tamaño de la población anual para cada área de estudio. Las tendencias representaron la proporción del tamaño de la población en el primer año que permaneció en cada año subsiguiente. De manera similar a λ_t, en la que se basaron, éstas Δ_t mostraron evidencia de disminución durante el período de estudio para LAS y SIE. El mejor modelo de reclutamiento indicado, el reclutamiento de búhos machos y hembras, adultos y subadultos, varió de 0.10 a 0.31 individuos territoriales nuevos al tiempo t por el número de individuos territoriales al tiempo t–1 y de manera similar entre las otras áreas. También realizamos un análisis de viabilidad de población (PVA) basado en los resultados de nuestro meta-análisis. Este análisis PVA fue de limitada utilidad para las áreas de estudio ELD y SKC porque el 95% de intervalos de confiabilidad en la probabilidad de disminución o aumento extendió el intervalo [0, 1] de 5–10 años. Cuando restringimos las inferencias a 7 años, la probabilidad estimada de a >10% de disminución para SIE fue 0.41 (95% CI = 0.09–0.78); para LAS la probabilidad fue 0.64 (95% CI = 0.27–0.94). En contraste, la probabilidad estimada de un >10% de aumento en 7 años para SIE fue 0.23 (95% CI = 0.01–0.55) y para LAS fue 0.10 (95% CI = 0.00–0.34). Para comparar, simulamos un PVA para una población hipotética con una media Λ = 1.0, y con la misma variación temporal observada en nuestras poblaciones de búhos. Nuestro PVA sugirió que ambas poblaciones SIE y LAS tenían, en un período de 7 años, mayores probabilidades de disminución que de aumento, pero que sería muy difícil determinar si alguna de las poblaciones estaba en una ligera disminución gradual. El depósito de información de nuestras 4 áreas de estudio provee una plantilla rica en información para que los administradores monitoreen los impactos de acciones futuras en el manejo de los búhos (por ejemplo, nuevas estrategias de manejo del Plan de Sierra Nevada Forest). También provee evidencia importante para evaluar el estatus del búho para su potencial inclusión en el listado de especies amenazadas. RÉSUMÉ Le hibou tacheté californien (Strix occidentalis occidentalis) est la seule sous-espèce de hibou tacheté ne figurant pas sur la liste des animaux menacés ou vulnérables sous la Loi des Espèces en Danger des Etats-Unis malgré des pétitions pour l'inscrire sur cette liste en tant que sous-espèce menacée. Nous avons effectué une méta-analyse des données de population pour 4 populations dans le sud des Cascades et dans la Sierra Nevada, en Californie de 1990 à 2005 pour aider une évaluation de leur statut établie par les Services des Eaux et Forêts des Etats-Unis. Nos aires d'étude (du nord au sud) étaient dans la forêt nationale Lassen (LAS), la forêt nationale Eldorado (ELD), la forêt nationale Sierra (SIE), et les parcs nationaux Sequoia et Kings Canyon (SKC). Ces aires d'étude représentaient un large échantillon des conditions de l'habitat et de la gestion dans ces chaînes de montagnes. Nous avons estimé la probabilité de survie apparente, le succès de reproduction, et le taux de changement de la population pour les hiboux tachetés dans chaque aire d'étude individuelle et dans toutes les aires réunies (méta–analyse) en utilisant la sélection de modèles ou le calcul de la moyenne des modèles basé sur une estimation du maximum de vraisemblance. Pour effectuer cette analyse nous avons suivi un protocole rigoureux similaire à d'autres méta-analyses de hiboux tachetés. La cohérence des observations de terrain et des méthodes analytiques entre ces études a réduit les effets méthodologiques confondants lors des évaluations des résultats. Nous avons utilisé 991 hiboux tachetés marqués dans l'analyse de survie apparente. La probabilité de survie apparente a été plus élevée pour les hiboux adultes que pour les sous-adultes. Il y a eu peu de différence pour ce qui est de la survie apparente entre les hiboux mâles et femelles. La moyenne des estimations de la probabilité de survie apparente des hiboux adultes basée sur la moyenne des modèles a varié entre 0,811 ± 0,021 pour les hiboux femelles à LAS et 0,890 ± 0,016 pour les hiboux mâles à SKC. La survie apparente a augmenté avec le temps pour les hiboux de toutes les classes d'âge à LAS et SIE, pour les adultes à ELD, et pour les sous-adultes de deux ans et les adultes à SKC. La méta-analyse de survie apparente, qui comprenait seulement des hiboux adultes, a confirmé une tendance croissante de survie avec le temps. Les taux de survie étaient plus élevés pour les hiboux de SKC que pour ceux des autres aires d'étude. Nous avons analysé les données obtenues à partir de 1 865 observations de succès de reproduction de hiboux tachetées femelles. La proportion des hiboux femelles sous-adultes parmi toutes les femelles territoriales d'àge connu a varié de 0,00 à 0,25 selon les aires et les années d'étude. La proportion des sousadultes parmi les hiboux tachetés femelles a été négativement corrélée avec le succès de reproduction (nombre de jeunes hiboux par femelle territoriale) pour ELD et SIE. La forêt nationale Eldorado et la forêt nationale Lassen ont montré une tendance à alterner selon un cycle biennal pour ce qui est du succès de reproduction, avec un taux plus élevé pendant les années paires. La moyenne du succès de reproduction annuel était de 0,988 ± 0,154 pour ELD, de 0,624 ± 0,140 pour LAS, de 0,478 ± 0,106 pour SIE, et de 0,555 ± 0,110 pour SKD. La forêt nationale Eldorado a montré une tendance décroissante ainsi que la plus grande variation dans le succès de reproduction avec le temps, alors que SIE et SKC, qui ont eu le succès de reproduction le plus bas, ont connu la variation temporelle la plus basse. La méta-analyse a confirmé que le succès de reproduction variait selon les aires d'étude. Le succès de reproduction a été le plus élevé pour les adultes, puis pour les sous-adultes de deux ans, et ensuite pour les sous-adultes d'un an. Nous avons utilisé 842 hiboux marqués, adultes et sous-adultes, pour estimer le taux de changement de la population. La modélisation a indiqué que Λ _t était soit stationnaire (LAS et SIE), soit croissant après une baisse initiale (ELD et SKC). La moyenne estimée Λ _t pour les 4 aires d'étude était: 1,007 (95% IC = 0,952–1,066) pour ELD; 0,973 (95% IC = 0,946–1,001) pour LAS; 0,992 (95% IC = 0,966–1,018) pour SIE; et 1,006 (95% IC = 0,947–1,068) pour SKC. Le meilleur modèle de méta-analyse pour la tendance de la population a indiqué que Λ variait selon le temps mais suivait la même tendance selon les aires d'étude. Nos estimations du changement de population réalisé (Δ _t ) étaient fondées sur les estimations de Λ _t des aires d'étude individuelles et n'ont pas nécessité d'estimation de la taille annuelle de la population pour chaque aire d'étude. Les tendances représentaient la proportion de la taille de la population pendant la première année qui s'est maintenue chaque année subséquente. De même que λ_t sur lesquels ils étaient fondés, ces δ_t ont apporté des preuves de déclin pendant la période d'étude pour LAS et SIE. Le meilleur modèle a indiqué que le recrutement des hiboux adultes et sous-adultes mâles et femelles variait de 0,10 à 0,31 nouveaux individus territoriaux à un temps t pour un nombre d'individus territoriaux à un temps t-1 et qu'il en était de même dans chaque aire. Nous avons également procédé à une analyse de viabilité de la population (AVP) fondée sur les résultats de notre méta-analyse. Cette AVP a été d'une utilité limitée pour les aires d'étude ELD et SKC parce que les intervalles de confiance de 95% sur la probabilité du déclin ou de la croissance couvraient l'intervalle [0, 1] sur une période de 5 à 10 ans. Lorsque nous avons réduit les inférences à 7 ans, la probabilité estimée d'un déclin >10% pour SIE était de 0,41 (95% IC = 0,09–0,78); pour LAS la probabilité était de 0,64 (95% IC = 0,27–0,94). Al'opposé, la probabilité estimée d'une croissance >10% en 7 ans pour SIE était de 0,23 (95% IC = 0,01–0,55) et pour LAS elle était de 0,10 (95% IC = 0,00–0,34). Afin de comparer, nous avons simulé une AVP pour une population hypothétique ayant une moyenne Λ = 1,0 et la même variation temporelle que celle observée dans nos populations de hiboux. Notre AVP a suggéré que les populations de SIE et de LAS avaient de plus grandes probabilités de déclin que de croissance sur une période de 7 ans, mais qu'il serait difficile de déterminer si une population présentait un léger déclin graduel. La collecte des informations pour nos 4 aires d'étude foumit aux personnes chargées de la gestion un modèle riche de données permettant de suivre l'impact sur les hiboux des actions de gestion à l'avenir (par exemple, les nouvelles stratégies de gestion du Plan pour la Forêt de Sierra Nevada). Cette collecte foumit également des preuves importantes afin d'évaluer le statut du hibou pour une classification potentielle sur la liste des espèces menacées.     

Uptake and metabolism of 6-benzylaminopurine in shoot cultures of

Shoots of Gerbera jamesonii Bolus, cultured in vitro were induced to multiply by the addition of 2.22 μ M 6-benzylaminopurine (BAP) to the medium. Shoots growing in the presence of [¹⁴C]-BAP were harvested every 4 days and sub-divided into petioles plus apices, laminae and basal callus. The 3- and 9-glucosides, 9-riboside and a novel compound, the 9-ribosylglucoside were characterised as metabolites of BAP by mass spectrometry. The 9-riboside was the principal metabolite formed initially in petioles plus apicies and callus but further metabolism varied depending on tissue type. After 24 days the major metabolite in the callus was the 9-ribosylglucoside, with only trace amounts of the other metabolites present. In contrast, both 3-glucoside and 9-ribosylglucoside accumulated in petioles plus apices whereas the 3-glucoside accumulated in laminae.     

Whole-exome sequencing identifies homozygous AFG3L2 mutations in a spastic ataxia-neuropathy syndrome linked to mitochondrial m-AAA proteases

We report an early onset spastic ataxia-neuropathy syndrome in two brothers of a consanguineous family characterized clinically by lower extremity spasticity, peripheral neuropathy, ptosis, oculomotor apraxia, dystonia, cerebellar atrophy, and progressive myoclonic epilepsy. Whole-exome sequencing identified a homozygous missense mutation (c.1847G>A; p.Y616C) in AFG3L2, encoding a subunit of an m-AAA protease. m-AAA proteases reside in the mitochondrial inner membrane and are responsible for removal of damaged or misfolded proteins and proteolytic activation of essential mitochondrial proteins. AFG3L2 forms either a homo-oligomeric isoenzyme or a hetero-oligomeric complex with paraplegin, a homologous protein mutated in hereditary spastic paraplegia type 7 (SPG7). Heterozygous loss-of-function mutations in AFG3L2 cause autosomal-dominant spinocerebellar ataxia type 28 (SCA28), a disorder whose phenotype is strikingly different from that of our patients. As defined in yeast complementation assays, the AFG3L2(Y616C) gene product is a hypomorphic variant that exhibited oligomerization defects in yeast as well as in patient fibroblasts. Specifically, the formation of AFG3L2(Y616C) complexes was impaired, both with itself and to a greater extent with paraplegin. This produced an early-onset clinical syndrome that combines the severe phenotypes of SPG7 and SCA28, in additional to other "mitochondrial" features such as oculomotor apraxia, extrapyramidal dysfunction, and myoclonic epilepsy. These findings expand the phenotype associated with AFG3L2 mutations and suggest that AFG3L2-related disease should be considered in the differential diagnosis of spastic ataxias.     

Natural and induced polyploidy in Acacia dealbata Link. and Acacia mangium Willd

Seeds were obtained from seven natural populations of Acacia dealbata, three natural populations of A. mangium and a seed orchard of A. mangium, representing the natural range of the two species. Polyploids were discovered in two of the seven populations of A. dealbata. The 2C DNA amount for diploid A. dealbata (2n = 2x = 26) was 1.74 pg, and for diploid A. mangium (2n = 2x = 26) was 1.30 pg. A naturally occurring tetraploid of A. dealbata (2n = 4x = 52) had a 2C DNA amount of 3.41 pg and a naturally occurring triploid genotype had a 2C DNA amount of 2.53 pg. The use of colchicine and oryzalin was investigated as a means of producing higher frequencies of tetraploids of both A. mangium and A. dealbata for incorporation into breeding programmes. Colchicine treatment gave tetraploid frequencies up to 29% for A. dealbata seedlings, and up to 18% for A. mangium seedlings. In contrast, no tetraploid A. mangium was detected following oryzalin treatment, and the low frequencies of tetraploids observed in A. dealbata could be attributed to their natural occurrence.     

Differential vulnerability of lambs to coyote predation

Coyote-lamb interactions in 65-ha pastures were observed during a study of Komondor dogs guarding sheep. Vulnerability of lambs to coyote predation was related to total time lambs were exposed to coyotes in the enclosures and total time present in the flock. Lambs were most vulnerable during the first two weeks of exposure (P < 0.001, X² = 49.3), and were somewhat less vulnerable in the next 4 weeks of exposure. If the lambs survived 6 weeks of exposure, their chances of succumbing to predation were small. Lambs were most vulnerable if they were added to an established flock. Orphan lambs were more vulnerable than ewe-reared lambs. Even though the coyotes in this study were persistent in attacking lambs, their success was limited for at least 3 reasons. First, the lambs sometimes refused to run from the coyotes. In addition, they occasionally actively defended themselves by foot-stomping, butting and approaching the coyote. Finally, no lambs were killed while in close proximity to a guardian dog. Based on this study, predation might best be reduced by minimizing introductions of new lambs, and especially orphan lambs, to established flocks, and by using a guardian dog, if possible.     

Angiotensin II receptors are reduced in the CNS of the young Brattleboro rat

In the rat, angiotensin II (AII), following specific interaction with sensitive central nervous system (CNS) receptors promotes release of vasopressin (ADH). We have examined the integrity of this chain of events by comparing the concentration, Bmax, and dissociation constant, Kd, in the CNS of Brattleboro rats (BB), a strain incapable of synthesizing ADH, with Long Evans (LE) control rats that can synthesize ADH. AII binding properties in the hypothalamic-thalamic-septal-midbrain (HTSM) area from young and old BB and LE, as well as systolic blood pressure, were determined. There was a reduction in the HTSM-AII receptor concentration of young BB when compared with young LE rats. Young BB had lower pressure than age and sex matched LE controls. Neither Bmax nor pressure was significantly different between older BB and LE. A decline in HTSM-AII receptor concentration with age observed with LE is consistent with observations in SHR and WKY rats. Parallel Scatchard plots obtained indicated the presence of a single class of CNS AII receptors. These data suggest that ADH synthesis and AII receptor concentration are partially interdependent and that the CNS AII-ADH system is redundant in the maintenance of blood pressure.     

A kinetic and structural characterization of adenosine-5'-triphosphate: ribonucleic acid adenylyltransferase from Pseudomonas putida.

A catalytic and structural study of ATP:RNA adenylyltransferase (EC 2.7.7.19) from the particulate fraction of Pseudomonas putida was made. During the large-scale purification of this enzyme, designated adenylyltransferase B, a previously undetected ATP-incorporating activity, designated adenylyltransferase A, was observed. Adenylyltransferases A and B were indistinguishable catalytically; however, they differed in their chromatographic and sedimentation properties. Adenylyltransferases A and B were resolved by phosphocellulose, by poly (U)-Sepharose and by Bio-Gel P-100 chromatographies. Adenylytransferase A was determined to have a sedimentation coefficient (S020,w) of 9.3 S and B of 4.3 S. The molecular weight of adenylyltransferase A was estimated to be 185000 and that of adenylyltransferase B to be 50000-60000. Apparently, adenylyltransferase A was generated from adenylyltransferase B during the purification. The AMP incorporation catalyzed by adenylyltransferases A and B was inhibited by two derivatives of the antibiotic rifamycin, AF/013 (50% at 5 mug/ml) and AF/DNFI (50% at 10 mug/ml). The 5'-triphosphate derivative (3'-dATP) of the drug cordycepin (3'-deoxyadenosine/ was a competitive inhibitor with ATP for both adenylyltransferases. The Ki for 3'-deoxyadenosine 5'-triphosphate was 6 - 10(-4)--10 - 10(-4) M, while the Km for ATP was 1 - 10(-4)--2 - 10(-4) M. Several other anaolgs of ATP, 2'-deoxyadenosine 5' triphosphate, 2'-O-methyl ATP, or the fluorescent 3-beta-D-ribofuranosylimidazo [2,1-i] purien 5'-triphosphate did not affect the activity of adenylyltransferase A or B. Poly(U) and poly(dT) were competitive inhibitors of the ribosomal RNA-primed polymerization reaction. The Ki for poly(U) or poly(dT), in terms of nucleotide phosphate, was 4 - 10-6)--10 - 10(-6) M for adenylyltransferases A and B, compared to 2 - 10(-4)--4 - 10(-4) M for the Km of ribosomal RNA. The inhibition was a result of the competition between the non-priming poly(U), or poly(dT), and ribosomal RNA for the primer binding site on the enzyme.