bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of parental age and diet of Dacus oleae Gmelin (Dipt., Tephritidae) on the biological performance of the filial generation
Authors: TSIROPOULOS G. J.
Published: July 2010

Parental age and diet affected the longevity, fecundity and fertility of the filial generation of Dacus oleae. The longevity of female offspring was shortened when the parental diet was sucrose. Fecundity and fertility was affected by both, parental age and parental diet. Females from young parents, up to 25 days old, had a higher egg production, while fertility was reduced when parental age was 50 days. Flies coming from sucrose‐fed parents produced significantly fewer eggs, than flies coming from parents fed the full diet. To maximize egg production in the insectary, flies coming from parents of age not more than 25‐day old should be used, provided that a full adult diet will be continuously available.

Zur Wirkung des Elternalters und der Nahrung von Dacus oleae Gmelin (Dipt., Tephritidae) auf die biologische Leistung der Tochtergeneration

Das Alter der Elterntiere sowie die Nahrung beeinflussen die Lebensdauer, Fruchtbarkeit und Eiablage der Tochtergeneration von D. oleae. Die Lebensdauer der weiblichen Nachkommen wurde verkärzt, wenn die Nahrung der Eltern aus Sucrose bestand. Fruchtbarkeit und Eiablage wurden von beiden Faktoren, Alter und Nahrung der Eltern beeinßt. Die Eiproduktion von Weibchen, die von 25 Tage alten Eltern stammten, war höher als jene von 50 Tage alten Eltern. Nachkommen von Eltern mit Sucrose‐Ernährung erzeugten signifikant weniger Eier als solche, deren Eltern eine Volldiät bekamen. Um im Insektarium die Eiproduktion zu maximieren, sollten Fliegen verwendet werden, deren Eltern nicht älter als 25 Tage waren und mit einer Volldiät ernährt wurden.
Pheromone 491: Freilandfänge von Geometriden‐Männchen mit C19‐Polyenen in Kiefernbeständen
Authors: BOGENSCHÜTZ H. , KNAUF W. , TRÖGER E. J. , BESTMANN H. J. , VOSTROWSKY O.
Published: July 2010

Pheromones 49: Males of Geometridae catched by C19‐Polyenes in pine stands

The polyenic C19‐hydrocarbons (6Z,9Z)‐6,9‐nonadecadiene, (3Z,6Z,9Z)‐nonadecatriene and (3Z,6Z,9Z)‐1,3,6,9‐nonadecatetraene were tested for sex attractant activity for male geometrid moths in pinewoods in Baden, Germany, and trap catches observed for eight different species of Boarmiinae, Larentiinae and Sterrhinae subfamilies. Despite of the low catch rates species specific attractancy of the nonadecadiene was found above all with Boarmia repandata and could be enhanced by addition of the triene or tetraene to the bait.

Die Polyen‐C19‐Kohlenwasserstoffe (6Z,9Z)‐6,9‐Nonadecadien, (3Z,6Z,9Z)‐3,6,9‐Nonadecatrien und (3Z,6Z,9Z)‐1,3,6,9‐Nonadecatetraen wurden in badischen Kiefefernbeständen als Sexuallockstoffe für Geometridenmännchen getestet, und die Anflüge bzw. Fallenfänge von 8 verschiedenen Arten aus den Unterfamilien Boarmilian, Larentiinae und Sterrhinae beobachtet. Trotz der niedrigen Fangzahlen wurde vor allem für das Nonadecadien eine artspezifische Lockwirkung gegenüber Boarmia repandata‐Männchen gefunden, die durch Zugabe des C19‐triens oder ‐tetraens noch erhöht wurde.
Über die Rolle von Pheroprax bei der Besiedlung der Fichte durch den Buchdrucker (Ips typographus L.)1
Authors: BOMBOSCH S. , ENGLER I. , GOSSENAUER H. , HERRMANN B.
Published: July 2010

On the role of Pheroprax influencing the settlement of Ips typographus L. on spruce The attractants of Pheroprax are effective only with flying bark‐beetles. A concentration gradient of their impact is observed. When combined with a clearly visible elongated silhouette they guide the beetles in a direct approach to the source of the attractant. In absence of this silhouette an attraction and landing of the beetles is still registered but the latter is directed much less distinctly to the source of the attractant.

In consequence it is assumed that further compounds hitherto unknown are responsible for the recognition of the host, the distribution on the trunk, and the finding of the mates.

Die im Pheroprax enthaltenen Lockstoffe wirken nur auf fliegende Buchdrucker. Ihre Wirkung ist konzentrationsabhängig. In Verbindung mit einer deutlich sichtbaren, langgestreckten Silhouettc führen sie zu gerichteten Anflügen der Duftstoffquelle. Fehlt diese Silhouette, kommt es auch zu Anlockung und Landung, jedoch ist diese wesentlich weniger auf die Duftquelle ausgerichtet. Danach müssen für die Wirtsfindung, die Verteilung am Stamm sowie das Auffinden der Geschlechter weitere, bisher nicht bekannte Verbindungen verantwortlich sein.
Atmospheric humidity and aphid reproduction
Authors: LEATHER SIMON R.
Published: July 2010

Apterous exules of the bird cherry‐oat aphid, Rhopalosiphum padi (L.) were significantly more fecund on oat seedlings at lower relative humidities (35% RH) than at higher relative humidities (55% RH). This was considered to be due to the increased transpiration rates of the plants under low humidity conditions. The significance of these results in relation to the practice of spring irrigation of cereals in Finland is discussed.

Luftfeuchtigkeit und Reproduktion bei Blattläusen

Ungeflügelte Exulantien der Vogelkirschen‐Hafer‐Blattlaus Rhopalosiphum padi (L.) waren sig‐nifikant fruchtbarer an Hafer‐Sämlingen bei niedriger (35 %) als höherer rel. Luftfeuchte (55 %). Es wird angenommen, daß dies auf der gesteigerten Transpirationsrate der Pflanze bei niedriger Luftfeuchte beruhte. Die Bedeutung dieser Ergebnisse in Bezug auf die Praxis von Frühjahrs‐Berieselungen bei Getreide in Finnland wird erörtert.
Zur Steuerung der Populationsdichte von phytophagen Insekten durch ihre Wirtspflanze
Authors: LUNDERSTÄDT J. , AHLERS SABINE
Published: July 2010

As to control of the population density of phytophagous insects by their food‐plant. 3. Control of the physiological performance Larvae of Gilpinia hercyniae were reared on spruce branches from the 13th day after hatching at 20°C, 18‐h‐photoperiod. The branches were harvested in May, July, and October respectively and were used as food source for the larvae at 25°C, 18‐h‐photoperiod until the beginning of the measurements.

From the 13th day onward the biological data were registered every second day until the imagines were developed, while the corresponding biochemical data were determined from the 13th day until pupation. On the basis of the rates of intake of 16 proteinbound amino acids and 9 non‐amino compounds, the control function for the regulation of the physiological performance of the insects in response to the changes of the external conditions was formulated which is valid for all three experiments. It is concluded from the sequence of the extrema of the rates of intake of quinic acid, shicimic acid, piceine, proteinbound valine and phenylalanine that they carry the information of the control process.

Larven von Gilpinia hercyniae wurden im Mai, Juli und Oktober ab 13. Tag nach Schlupf bei 20°C, 18‐h‐Tag, an Fichtenäste angesetzt, die bei 25°C, 18‐h‐Tag, von frischgeschlüpften Larven bis zum 13. Tag vorbefressen worden waren. In zweitägigem Abstand wurden vom 13. Lebenstag an die biologischen Daten und vom 13. Lebenstag bis zur Verpuppung die zugehörigen biochemischen Daten erfaßt. Über die Aufnahmerate von 16 Proteinaminosäuren und 9 Nichtaminoverbindungen wird die Einsteuerung des Regelintervalls in Form einer für alle 3 untersuchten Fälle gültigen Steuergleichung angegeben, gemäß der die physiologische Leistung der Insekten auf sich ändernde Außenbedingungen eingestellt wird. Aus der zeitlichen Abfolge der Extrema der Aufnahmeraten von Chinasäure, Catechin, Picein sowie proteingebundenem Valin und Phenylalanin wird geschlossen, daß sie Informationsträger des Steuerungsprozesses sind.
Proof‐of‐concept trial on the efficacy of sodium tungstate in human obesity
Authors: Hanzu F. , Gomis R. , Coves M. J. , Viaplana J. , Palomo M. , Andreu A. , Szpunar J. , Vidal J.
Published: July 2010

Aim: Considering the poor long‐term success of current dietary and pharmacological interventions, we aimed to evaluate the potential effect of sodium tungstate in the treatment of grade I and II obesity (ClinicalTrials.gov identifier: NCT00555074).

Methods: Prospective, randomized, placebo‐controlled, double‐blind, proof‐of‐concept study was carried out. Following a 2‐week lead‐in period, 30 obese (body mass index, BMI 30.0–39.9 kg/m2), non‐diabetic subjects were randomized to receive either sodium tungstate (100 mg bid) or placebo for 6 weeks. The primary study endpoint was the absolute change in body weight relative to the time of randomization.

Results: Treatment with sodium tungstate [−0.135 ± 0.268 kg (95% CI −0.686 to +0.416 kg)] was not associated with a significant weight loss compared to placebo [−0.063 ± 0.277 kg (95% CI −0.632 to +0.507 kg)] (p = 0.854). Likewise, treatment with sodium tungstate was not associated with significant changes in fat mass (DEXA), resting energy expenditure (indirect calorimetry) or caloric consumption (3‐day food records).

Conclusion: Our data do not support sodium tungstate as a pharmacological agent in the treatment of human obesity.
Ways of seeing environmental change: Participatory research engagement in Yunnan, China, with ethnic minority Hani participants
Authors: McKinnon John
Published: July 2010

In this article, I reflect on a participatory learning and action (PLA) and participatory geographic information system (PGIS) project undertaken in two adjacent Hani villages in southern Yunnan, China. After a lengthy process to gain research authorisations, Hani villagers worked with a group of visiting researchers (Hani, Han Chinese and New Zealanders) to articulate local visions of land‐use change and environmental challenges. PLA exercises produced a diverse range of hand‐made and PGIS products over a 10‐week period. As villagers became more accustomed to the ‘outsider team’, methods – both designed and spontaneous – diversified. Afterwards, based on the results and acting on their own initiative, local leaders backed by popular support moved quickly to reassert traditional and ritually prescribed Hani conservation measures. Nevertheless, these endogenously driven solutions were not necessarily universally beneficial. In describing the complexities encountered in implementing a participatory framework and the ambiguities of the outcomes engendered, I argue that privileging local coherence and celebrating such participatory approaches should not be done at the expense of ignoring the intricacies of on‐going contradictory behaviours in a rapidly changing arena. Yet, in China, where authorities often remain suspicious of those seeking to undertake long‐term fieldwork, especially among ethnic minorities, PLA offers a potential route forward.
PRECONVULSIVE CHANGES IN BRAIN GLUCOSE METABOLISM FOLLOWING DRUGS INHIBITING GLUTAMATE DECARBOXYLASE
Authors: Horton R. W. , Meldrum B. S.
Published: July 2010

—Brain glucose and glycogen concentrations have been studied in mice treated with allylglycine, 4‐deoxypyridoxine and isoniazid, and the effects compared with the preconvulsive increase in brain glucose and glycogen concentration that follows d, l‐methionine sulphoximine treatment. Allylglycine (180 mg/kg), 4‐deoxypyridoxine (250 mg/kg), isoniazid (150 mg/kg) and d,l‐methionine sulphoximine (300 mg/kg) when given to mice at room temperature, cause a fall in rectal temperature which can be prevented by maintaining the mice in an incubator at 33‐34°C. An increase in brain glucose concentration is seen after allylglycine (+ 133%), d,l‐methionine sulphoximine (+ 113%) and 4‐deoxypyridoxine (+ 70%) treatment when mice are kept at room temperature and killed before convulsions occur. This is associated with a rise in blood glucose concentration after allylglycine, but not after the other drugs. Preventing the fall in rectal temperature reduces, but does not abolish, the rise in brain glucose concentration seen after allylglycine, d,l‐methionine sulphoximine and 4‐deoxypyridoxine. Brain glycogen concentration increases at room temperature after D,L‐methionine sulphoximine and 4‐deoxypyridoxine, but in mice with maintained body temperature only 4‐deoxypyridoxine produces an increase in brain glycogen. Isoniazid does not increase brain glucose or glycogen at room temperature, but reduces their concentration in mice kept in the incubator. All four drugs are known to act on amino acid metabolism; d,l‐methionine sulphoximine potently inhibits glutamine synthetase whereas 4‐deoxypyridoxine, allylglycine and isoniazid inhibit glutamate decarboxylase. The connection, if any, between a block in the further metabolism of glutamate and an increase in brain glucose and glycogen is unknown.
A RADIOIMMUNOASSAY FOR MEASURING 14‐3‐2 PROTEIN IN CELL EXTRACTS
Authors: Revoltella R. , Bertolini L. , Diamond L. , Vigneti E. , Grasso A.
Published: July 2010

—A rapid and sensitive radioimmunoassay for the neuron‐specific protein, 14‐3‐2, has been developed. The assay was used to determine the amounts of 14‐3‐2 protein in soluble extracts prepared from clones of the C 1300 murine neuroblastoma and from C 1300 × L cell hybrid clones. The values obtained with the radioimmune procedure correlated with those obtained in microcomplement fixation and microprecipitin assays.
Aging Decreases the Sensitivity of Rat Cortical Synaptosomes to Calcium Ionophore‐Induced Acetylcholine Release
Authors: Meyer E. M. , Crews F. T. , Otero D. H. , Larsen K.
Published: July 2010

Abstract The capacity of calcium ions to trigger acetylcholine release was studied in cerebral cortical synaptosomes from adult (6‐month‐old) and senescent (24‐month‐old) rats, using a calcium ionophore, A23187, that bypasses voltage‐sensitive calcium channels. The potency but not the efficacy of the A23187 was reduced with respect to releasing acetylcholine (ACh) in the aged animals. There was no age‐related difference in the synthesis of ACh or potency of the ionophore with respect to increasing 45calcium uptake. These results suggest that aging reduces the sensitivity of cerebral cortical nerve terminals to calcium‐triggered ACh‐release.
Dopamine Metabolism in Hypoxanthine‐Guanine Phosphoribosyltransferase‐Deficient Variants of PC12 Cells
Authors: Bitler Catherine M. , Howard Bruce D.
Published: July 2010

Lesch‐Nyhan syndrome results from a deficiency of hypoxanthine‐guanine phosphoribosyltrans‐ferase (HPRT). It is manifest by behavioral abnormalities, including self‐mutilation, and evidence of abnormal 3,4‐dihydroxyphenylethylamine (dopamine) metabolism. To assess whether an HPRT deficiency in a dopaminergic cell can adversely affect dopamine metabolism in that cell, dopamine metabolism was examined in HPRT‐defi‐cient variants of PC12 pheochromocytoma cells and in cells that had regained HPRT activity by virtue of transformation with a recombinant retrovirus containing the human gene for HPRT. There was no correlation between HPRT activity and endogenous dopamine levels, dopamine uptake, dopamine release, or monoamine oxidase activity. Transformation with the HPRT retrovirus did not adversely affect dopamine metabolism.
Conversion of Leumorphin (Dynorphin B‐29) to Dynorphin B and Dynorphin B‐14 by Thiol Protease Activity
Authors: Devi Lakshmi , Goldstein Avram
Published: July 2010

Dynorphin B (rimorphin) is formed from leumorphin (dynorphin B‐29) by the action of a thiol protease from rat brain membranes, in a single step. This represents a “single‐arginine cleavage” between threonine‐13 and arginine‐14 of the substrate. We have observed that in addition to dynorphin B, dynorphin B‐14 is formed from dynorphin B‐29. Among the various protease inhibitors tested, none except p‐chloromercuriben‐zensulfonic acid inhibited the formation of the two products. Both temperature and pH had similar effects on the formation of dynorphin B‐14 and dynorphin B. The inhibitory potencies of adrenocorticotropic hormone, peptide E, and dynorphin A were virtually identical for the formation of the two products. These results suggest that the same enzyme may be responsible for the formation of dynorphin B‐14 and dynorphin B.
Extracellular Overflow of Neuroactive Amino Acids During Severe Insulin‐Induced Hypoglycemia: In Vivo Dialysis of the Rat Hippocampus
Authors: Sandberg Mats , Butcher Steven P. , Hagberg Henrik
Published: July 2010

Hypoglycemia‐evoked changes in levels of extracellular excitatory and inhibitory amino acids were studied using the microdialysis technique. A newly designed dialysis probe was inserted stereotaxically into the rat hippocampus. Animals were then subjected to insulin‐induced hypoglycemia; then blood glucose levels were restored by glucose injections after a 30‐min period of isoelectric electroencephalography. Dialysates were collected before, during, and after the isoelectric period. Amino acids in the dialysates were analyzed by liquid chromatography and fluorescence detection following automatic precolumn derivatization with o‐phthaldialdehyde. During the isoelectric phase, the concentration of aspartate increased 15‐fold, whereas glutamate, γ‐aminobutyric acid, taurine, and phosphoethanolamine levels were elevated three‐ to sixfold. Smaller increases were observed for nonneuroactive amino acids such as asparagine, alanine, and phenylalanine. In contrast to all other amino acids, the glutamine content was reduced to <30% of preisoelectric values. The concentrations of the neuroactive amino acids were restored to normal in the postisoelectric phase. These data demonstrate that there is an extracellular overflow of neuroactive amino acids, especially aspartate, during severe hypoglycemia.
Effect of CDP‐Choline on Hypocapnic Neurons in Culture
Authors: Mykita S. , Golly F. , Dreyfus H. , Freysz L. , Massarelli R.
Published: July 2010

Neuronal cultures from chick embryo cerebral hemispheres were protected against a hypocapnic injury by adding to their growth medium 10‐‐6M CDP‐choline before or after the injury. The protection obtained with CDP‐choline was analyzed by a morphometric analysis and showed that pretreatment of neuronal cultures with CDP‐choline maintained the number of cell aggregates and of primary neuronal processes at control values after hypocapnic shock. Various experiments showed that the intact molecule was responsible for the protective action, since pretreatment with different concentrations of various nucleosides and nucleotides (up to 10‐‐5M), choline, and phosphorylcholine was without protective effect. The addition of CDP‐choline after the hypocapnic injury resulted in a protection of the cultures as shown by morphological observation. Incubation of neurons with radioactive choline showed that hypocapnia increased the incorporation of the label into phospholipids whereas the presence of CDP‐choline reduced it. The de novo synthesis of choline was affected by neither hypocapnia nor CDP‐choline treatment. The results indicate that CDP‐choline may have the capacity to protect neurons under conditions of basic pH and that cellular proliferation may be stimulated by the compound.
Development and Characterization of Pantothenic Acid Transport in Brain
Authors: Spector Reynold
Published: July 2010

Abstract: In vitro, the transport of [3H]pantothenic acid into and from rabbit brain slices was studied. In newborn rabbits and throughout development, forebrain and cerebellar slices were able to accumulate and phosphorylate PHJpantothenic acid comparably to slices from adults. The accumulation and phosphorylation of [3H]pantothenic acid by adult forebrain slices were not decreased by substitution of LiCl for NaCl in the artificial CSF or by addition of short‐chain fuels (e.g., 5 mM pyruvate or acetoacetate) to the medium. However, probenecid and ouabain (both 1 mM) and medium‐chain fatty acids (e.g., 0.1 mM octanoate, nonanoate, and decanoate) profoundly inhibited [3H]pantothenic acid accumulation by forebrain slices but not intracellular phosphorylation and conversion to [3H]CoA. These in vitro results suggest that brain slices accumulate pantothenic acid by a saturable system (probably facilitated diffusion) that is sensitive to inhibition by probenecid and medium‐chain fatty acids.
Inhibition of Monoamine Oxidase by 3,4‐Dihydroxyphenylserine
Authors: Naoi Makoto , Nagatsu Toshiharu
Published: July 2010

Abstract:The effects of diastereomers of 3,4‐dihydroxyphenylserine (DOPS) on the enzyme activity of monoamine oxidase (MAO) in human placenta and liver mitochondria were examined. Both l‐ and d‐threo‐DOPS were found to inhibit MAO‐A in human placental mitochondria in competition with the substrate, and theKivalues for l‐ and d‐threo‐DOPS obtained were 68.3 and 125 μM, respectively. The inhibitory effect of l‐threo‐DOPS on both MAO‐A and ‐B activity was confirmed in human liver mitochondria, and MAO‐A was found to be more sensitive to the inhibitor. Other isomers of DOPS, l‐ and d‐erythro‐DOPS, were found to inhibit MAO activity, but the inhibition was noncompetitive with the substrate. The inhibitory effects of DOPS isomers were not affected by the presence of NSD‐1055, an inhibitor of aromaticl‐Amino acid decarboxylase, suggesting that the inhibition is the direct effect of DOPS, and not of norepinephrine produced by the decarboxylase.
Analysis of the Primary Sequence of Human Myelin Basic Protein Peptides 1–44 and 90–170 by Fast Atom Bombardment Mass Spectrometry
Authors: Scoble Hubert A. , Whitaker John N. , Biemann K.
Published: July 2010

Abstract: The originally described sequence of human myelin basic protein peptide 45–89 has recently been shown to contain two errors which have now been resolved. In the present study fast atom bombardment mass spectrometry was utilized to analyze the primary sequence of the other portions, peptides 1–44 and 90–170 of human myelin basic protein. The results obtained confirm the accuracy of the primary sequence published for both of these terminal peptides.
Membrane Potential Measured During Potassium‐Evoked Release of Noradrenaline from Rat Brain Neurons: Effects of Normorphine
Authors: Bug William , Williams John T. , North R. A.
Published: July 2010

Abstract:A single slice of rat pons that contained the locus ceruleus (LC) or two slices of cerebellum were loaded with [3H]noradrenaline; superfusion with high (35 or 60 mM) potassium solutions evoked a release of3H. In the presence of normorphine, the release of3H evoked by 35 mMpotassium and 60 mMpotassium was reduced. In some of those experiments in which the release of3H from the LC slice was measured, an intracellular microelectrode was used to measure membrane potential. This showed that solutions of increased potassium concentration depolarized the neurons to a potential at which inward calcium currents flowed (calcium action potentials occurred). Normorphine hyperpolarized the neurons; during this hyperpolarization the depolarization caused by 35 mMpotassium did not reach the threshold for significant calcium entry. The results suggest that the inhibition by normorphine of transmitter release evoked by solutions of raised potassium concentration could result in part from the membrane hyperpolarization caused by the normorphine.
A Neurotrophic Factor Derived from Goldfish Brain: Characterization and Purification
Authors: Mizrachi Yaffa , Rubinstein Menachem , Kimhi Yosef , Schwartz Michal
Published: July 2010

Previous studies carried out in our laboratory have demonstrated that goldfish brain contains substances that promote neurite extension from regenerating retinae in culture. Fractionation of the brain extract by molecular sieving chromatography revealed the presence of several molecular species, including two peaks that have neurotrophic activity, representing low‐molecular‐weight substances. One peak was eluted (P‐a) with an apparent molecular weight of about 13 kDa and was designated substratum neurite extension factor (SNEF) because it retained its neurotrophic activity when adsorbed onto the substratum. This recovered Sephadex fraction (P‐a) when applied in vivo intraocularly caused an earlier capacity of the corresponding retinae to sprout in vitro. Thus, at 3 and 5 days after injury the neuritic growth indices from the factor‐treated retinae were of 0.9 ± 0.2 and 2.8 ± 0.5, respectively, as compared with indices of 0.3 ± 0.1 and 0.9 ± 0.2, respectively, in retinae of injured but nontreated nerves. The factor was further purified by two steps of HPLC (ion exchange followed by reversed phase). The results showed that it is an acidic glycoprotein with an apparent molecular weight of 10 kDa.
Depolarization‐Dependent Protein Phosphorylation in Rat Cortical Synaptosomes: Characterization of Active Protein Kinases by Phosphopeptide Analysis of Substrates
Authors: Dunkley Peter R. , Baker Christine M. , Robinson Philip J.
Published: July 2010

Depolarization of synaptosomes is known to cause a calcium‐dependent increase in the phosphorylation of a number of proteins. It was the aim of this study to determine which protein kinases are activated on depolarization by analyzing the incorporation of 32P1 into synaptosomal phosphoproteins and phosphopeptides. The following well‐characterized phosphoproteins were chosen for study: phosphoprotein “87K,” synapsin Ia and Ib, phosphoproteins IIIa and IIIb, the catalytic subunits of calmodulin kinase II, and the B‐50 protein. Each was initially identified as a phosphoprotein in lysed synaptosomes after incubation with [γ‐32P]ATP. Mobility on two‐dimensional polyacrylamide gels and phosphorylation by specific protein kinases were the primary criteria used for identification. A technique was developed that allowed simultaneous analysis of the phosphopeptides derived from all of these proteins. Phosphopeptides were characterized in lysed synaptosomes after activating cyclic AMP‐, calmodulin‐, and phospholipid‐stimulated protein kinases in the presence of [γ‐32P]ATP. Phosphoproteins labelled in intact synaptosomes after incubation with 32Pi were then compared with those seen after ATP‐labelling of lysed synaptosomes. As expected from previous work, phosphoprotein “87K,” and synapsin Ia and Ib were‐labelled, but for the first time, phosphoproteins IIIa, IIIb, and the B‐50 protein were identified as being labelled in intact synaptosomes; the calmodulin kinase II subunits were hardly phosphorylated. From a comparison of the phosphopeptide profiles it was found that cyclic AMP‐, calmodulin‐, and phospholipidstimulated protein kinases are all active in intact synaptosomes and their activity is dependent on extrasynaptosomal calcium. The activation of cyclic AMP‐stimulated protein kinases in intact synaptosomes was confirmed by the addition of dibutryl cyclic AMP and theophylline which specifically increased the labelling of phosphopeptides in synapsin Ia and Ib and in phosphoproteins IIIa and IIIb. On depolarization of intact synaptosomes, a number of phosphopeptides showed increased labelling and the pattern suggested that cyclic AMP‐, calmodulin‐, and phospholipid‐stimulated protein kinases were all activated. No new peptides were phosphorylated, suggesting that depolarization simply increased the activity of already active protein kinases and that there was no depolarization‐specific increase in protein phosphorylation.
Mononucleotide Metabolism in the Rat Brain After Transient Ischemia
Authors: Onodera Hiroshi , Iijima Kunihiro , Kogure Kyuya
Published: July 2010

Nucleotide metabolism was studied in rats during and following the induction of 10 min of forebrain ischemia (four‐vessel occlusion model). Purine and pyrimidine nucleotides, nucleosides, and bases in forebrain extracts were quantitated by HPLC with an ultraviolet detector. Ischemia resulted in a severe reduction in the concentration of nucleoside triphosphates (ATP, GTP, UTP, and CTP) and an increase in the concentration of AMP, IMP, adenosine, inosine, hypoxanthine, and guanosine. During the recovery period, both the phosphocreatine level and adenylate energy charge were rapidly and completely restored to the normal range. ATP was only 78% of the control value at 180 min after ischemic reperfusion. Levels of nucleosides and bases were elevated during ischemia but decreased to values close to those of control animals following recirculation. Both the decrease in the adenine nucleotide pool and the incomplete ATP recovery were caused by insufficient reutilization of hypoxanthine via the purine salvage system. The content of cyclic AMP, which transiently accumulated during the early recirculation period, returned to the control level, paralleling the decrease of adenosine concentration, which suggested that adenylate cyclase activity during reperfusion is modulated by adenosine A2 receptors. The recovery of CTP was slow but greater than that of ATP, GTP, and UTP. The GTP/GDP ratio was higher than that of the control animals following recirculation.
Transport of Prostaglandins and Other Eicosanoids by the Choroid Plexus: Its Characterization and Physiological Significance
Authors: DiBenedetto F. E. , Bito L. Z.
Published: July 2010

Choroid plexi from the lateral ventricles of rabbits, cats, and dogfish (Mustelus canis) were used to characterize the prostaglandin (PG) uptake process and to establish its kinetic parameters and substrate specificity. The apparentKtfor PGF2αtransport by the rabbit choroid plexus was 20 μM; theJmaxwas 27 nmol g−1mm−1. TheKiof inhibition of PGF2αtransport by PGE2was 20 μM; theJmaxof PGF2αtransport was unaltered by PGE2. A concentration ofp‐aminohippuric acid of up to 1 mMdid not appreciably affect theK1or theJmaxof PGF2αtransport. The rate of PGF2αaccumulation by rabbit choroid plexus was reduced by incubation at 4°C, under anaerobic conditions, in the absence of sodium or in the presence of ouabain, probenecid, or bromcresol green. The choroid plexi of all three species also accumulated thromboxane B2, PGI2, and 6‐keto‐PGF1α, suggesting that most, if not all, eicosanoids are substrates for this transport system. It is concluded that the choroid plexus transport system satisfies all the criteria of an active, energy‐dependent transport system and that this system functions effectively at concentrations of eicosanoids present in the ventricular system under normal or pathological conditions. Hence, this transport system must make an important contribution to the pharmacokinetics of eicosanoids within the brain.
Subdivision of Mouse Brain [3H]Imipramine Binding Based on Ion Dependence and Serotonin Sensitivity
Authors: Severson James A. , Woodward John J. , Wilcox Richard E.
Published: July 2010

The specific binding of [3H]imipramine to mouse brain membranes in an assay containing 120 mMNaCl and 5 mMKCl was similar in regional distribution and pharmacological specificity to that reported previously in rat and human brain. However, the absence of ions decreased the density of the specific binding of [3H]imipramine and did not affect the equilibrium dissociation constant. Sodium was the only cation, and halides were the only anions tested that enhanced the specific binding of [3H]imipramine. Chloride did not increase the density of binding in the absence of sodium. The ion‐sensitive binding of [3H]imipramine was regionally dependent and was highly correlated with the uptake of 5‐hydroxytryptamine (5‐HT, serotonin) into synaptosomes from brain regions. 5‐HT did not inhibit the binding of [3H]imipramine in the absence of ions. Antidepressants inhibited binding in the absence and presence of ions, but in the presence of ions inhibition curves were shifted to the left and the apparent complexity of inhibition was increased. Quantitative analysis of the inhibition of [3H]imipramine binding by antidepressants conducted in the presence of ions was consistent with two binding sites. Lesion of the serotonergic input to the cerebral cortex by 5,7‐dihydroxytryptamine suggested that both the 5‐HT‐sensitive and ion‐sensitive binding of [3H]imipramine were associated with serotonergic nerve terminals. [3H]Imipramine binding displaced by desipramine, but insensitive to 5‐HT and ions, was not affected by the lesion. Thus, the binding of [2H]imipramine that is displaced by desipramine, the most common assay for [3H]imipramine binding, includes a component that is not associated with brain serotonergic nerve terminals and 5‐HT uptake, and, in addition, a separable component that is highly correlated with serotonergic function. These data have important implications for studies of serotonergic neurons and for the interpretation of imipramine binding data.
GABABReceptor‐Mediated Enhancement of Vasoactive Intestinal Peptide‐Stimulated Cyclic AMP Production in Slices of Rat Cerebral Cortex
Authors: Watling Keith J. , Bristow David R.
Published: July 2010

Basal and vasoactive intestinal peptide (VIP)‐stimulated accumulations of cyclic AMP were measured in slices of rat cerebral cortex. Neither γ‐aminobutyric acid (GABA) nor the selective GABABreceptor agonist (–)‐baclofen stimulated basal cyclic AMP accumulation, whereas VIP caused a large dose‐dependent increase in cyclic AMP levels. However, in the presence of 100 μM(–)‐baclofen, the effects of VIP on cyclic AMP accumulation were significantly enhanced, with the responses to 1 μMand 10 μMVIP being approximately doubled. The enhancing effects of (–)‐baclofen was dose related (1–1,000 μM), but an enhancing effect was not observed with 100 μM(+)‐baclofen. In the presence of the GABA uptake inhibitor nipecotic acid (1 mM), GABA caused a similar dose‐related enhancement of the VIP response. The ability of either GABA or (–)‐baclofen to augment VIP‐stimulated production of cyclic AMP was not mimicked by the GABAAagonists isoguvacine and 4,5,6,7‐ tetrahydroisoxazolo[5,4‐c]pyridin‐3‐ol (THIP) and was not antagonized by the GABAAantagonist bicuculline. The putative GABABantagonist 5‐aminovaleric acid (1 mM) significantly reduced the effect of (–)‐baclofen. The ability of (–)‐baclofen to enhance VIP‐stimulated accumulation of cyclic AMP was observed in slices of rat cerebral cortex, hippocampus, and hypothalamus. These results indicate that GABA and (–)‐baclofen can enhance VIP‐stimulated accumulation of cyclic AMP in rat brain slices via an interaction with specific GABABreceptors.
Stimulatory Effects of Exogenous Thyroid Hormone and Growth Hormone on sn‐Glycerol‐3‐Phosphate Dehydrogenase Activity in the Genetic‐Hypothyroid Mutant Mouse Cerebellum: Restricted to the Second 20 Days of Postnatal Life
Authors: Sugisaki Tetsuro , Noguchi Tetsuya
Published: July 2010

Abstract: We recently reported that by postnatal day 40 the activity of sn‐glycerol‐3‐phosphate dehydrogenase (GPDH) was significantly depressed in the cerebellum of genetic‐hypothyroid mutant mice. This mutant mouse‐GPDH combination was used in the present study to define the critical time period during which thyroid hormone (T4) and growth hormone (GH) are essential for maturation of Bergmann glial cells. Our findings are that (a) induction of GPDH activity in the Bergmann glial cell is dependent on T4, (b) T4 is most effective when administered during the second 20 days of postnatal life, (c) the effect of GH on GPDH activity is complementary to or synergistic with that of T4, and (d) Bergmann glial cells and radial glial fibers of the mutant mice contain immunoreactive GPDH following various hormonal treatments. These results suggest that T4 is indispensable for the maturation of Bergmann glial cells.
Neomycin Is an Agonist at a Polyamine Site on the N‐Methyl‐D‐Aspartate Receptor
Authors: Pullan L. M. , Stumpo R. J. , Powel R. J. , Paschetto K. A. , Britt M.
Published: July 2010

Abstract:Neomycin appears as a full agonist and spermidine as a partial agonist at the site where polyamines enhance 1‐[1‐(2‐thienyl)cyclohexyl][3H]piperidine ([3H]TCP) binding on theN‐methyl‐D‐aspartate (NMDA) receptor. Other aminoglycosides also enhance [3H]TCP binding with efficacies roughly proportional to the number of primary amine groups. The polyamine antagonists ifenprodil and arcaine inhibit enhancement of [3H]TCP binding by spermidine or neomycin. The inhibition of [3H]TCP binding by arcaine is apparently competitively reduced by neomycin and spermidine, supporting a common site. Diethylenetriamine (previously described as a polyamine antagonist) may be a partial agonist. Enhancement by neomycin or spermidine is not additive to that of Mg2+, consistent with competition of Mg2+and spermidine or neomycin at the site where these compounds enhance [3H]TCP binding. Polyamines also enhance the binding of the competitive antagonist 2 ‐ (2 ‐ carboxypiperazin ‐ 4 ‐ yl)[3H]propyl ‐ 1 ‐ phosphonic acid ([3H]CPP). Neomycin, which does not enhance [3H]CPP binding, inhibits the enhancement by spermidine. That this site is distinct from the site where spermidine and neomycin increase [3H]TCP binding is supported by different pharmacology. Arcaine and diethylenetriamine do not inhibit spermidine enhancement of [3H]CPP binding. Mg2+also does not compete with the spermidine enhancement of [3H]CPP binding. Ifenprodil inhibits the spermidine enhancement of [3H]CPP binding. The data suggest two or more polyamine sites, with arcaine selective for the site that enhances [3H]TCP binding. Neomycin is an agonist at one polyamine site and an antagonist to the second.
Extracellular Striatal Concentrations of Endogenous 3,4‐Dihydroxyphenylalanine in the Absence of a Decarboxylase Inhibitor: A Dynamic Index of Dopamine Synthesis In Vivo
Authors: Carboni Ezio , Tanda Gianluigi , Di Chiara Gaetano
Published: July 2010

Abstract:Basal levels of endogenous 3,4‐dihydroxyphenylalanine (DOPA) were detected by HPLC coupled with coulometric detection in dialysates from freely moving rats implanted 48–72 h earlier with transversal dialysis fibers in the dorsal caudate. Because decarboxylase inhibitor is absent in the Ringer's solution, this method allows monitoring of basal output of dopamine (DA) and 3,4‐dihydroxyphenylacetic acid, as well as DOPA. Extracellular DOPA concentrations were reduced by the tyrosine hydroxylase inhibitor α‐methylparatyrosine (200 mg/kg, i.p.) and by the dopaminergic agonist apomorphine (0.25 mg/kg, s.c.). The dopaminergic antagonist haloperidol (0.2 mg/kg, s.c.) stimulated DOPA output by about 60% over basal values. γ‐Butyrolactone, at doses of 700 mg/kg, i.p., which are known to block dopaminergic neuronal firing and which reduce DA release, stimulated DOPA output maximally by 130% over basal values. Tetrodotoxin, which blocks DA release by blocking voltage‐dependent Na+channels, increased DOPA output maximally by 100% over basal values. The results indicate that basal DOPA can be detected and monitored in the extracellular fluid of the caudate of freely moving rats by transcerebral dialysis and can be taken as a dynamic index of DA synthesis in pharmacological conditions.
Alteration of Neuroblastoma Ganglioside Metabolism by Retinoic Acid
Authors: Li Ruixiang , Ladisch Stephan
Published: July 2010

Abstract: Cellular differentiation is often associated with striking changes in ganglioside metabolism. Because retinoic acid causes cellular differentiation in vitro, we have characterized its effect on ganglioside synthesis and shedding by LAN‐5 human neuroblastoma cells. Three major observations were made: (a) 20 μM retinoic acid caused a marked (twofold) increase in cellular ganglioside content, with a slight relative enhancement in GD1a and GT1b synthesis, (b) ganglioside shedding increased in parallel with increased cellular ganglioside content, and also, unexpectedly, (c) retinoic acid caused a quantitatively similar increase in content of cell membrane phospholipids, which are also shed. We conclude that enhanced ganglioside synthesis and shedding by retinoic acid are part of a previously undescribed generalized stimulatory effect on membrane lipid metabolism.
A Novel Transcript Overlapping the Myelin Basic Protein Gene
Authors: Grima Brigitte , Zelenika Diana , Pessac Bernard
Published: July 2010

Abstract: Myelin basic protein (MBP) is a major constituent of myelin synthesized by oligodendrocytes and Schwann cells. We have investigated the expression of mouse MBP RNAs outside the nervous system. Nuclease protection experiments indicate that RNAs containing exon 1 and not the six downstream exons of the MBP gene are transcribed in various hemopoietic tissues. We have isolated a hemopoietic MBP‐related (HMBPR) cDNA clone from a mouse bone marrow cDNA library screened with an MBP cDNA probe. This clone contains exons 1a and 1b and a part of intron 1 of the MBP gene. An additional 5’region, encoded by at least three unidentified exons, lies upstream of exon 1a. The HMBPR clone corresponds to a 5‐kb RNA expressed in bone marrow, spleen, thymus, and macrophagic cells. This transcript is expressed at a similar level in brain, although at a lower level than the classical 2‐kb mRNA. These data indicate that a new transcript, overlapping the MBP transcription unit and controlled by a distinct promoter, is expressed in hemopoietic tissues. This RNA might encode a 21‐kDa protein sharing a common domain with MBP.
Lithium Enhances Accumulation of [3H]Inositol Radioactivity and Mass of Second Messenger Inositol 1,4,5‐Trisphosphate in Monkey Cerebral Cortex Slices
Authors: Dixon John F. , Lee Chang Ho , Los Georgyi V. , Hokin Lowell E.
Published: July 2010

Abstract: We previously reported that lithium, in the presence of acetylcholine, increased accumulations of inositol 1,4,5‐trisphosphate and inositol 1,3,4,5‐tetrakisphosphate in brain cortex slices from the guinea pig, rabbit, rat, and mouse. In the mouse and rat, the Li+‐induced increases required supplementation of the medium with inositol. This probably relates to the following facts: (a) Brain cortices of the mouse and rat contain in vivo concentrations of inositol half of that of the guinea pig. (b) Incubated rat brain cortex slices are depleted of inositol by 80%. (c) The slices require 10 mM inositol supplementation to restore in vivo concentrations. We now show that in monkey brain cortex slices, therapeutic concentrations of Li+ increase accumulation of inositol 1,4,5‐trisphosphate. The inositol 1,3,4,5‐tetrakisphosphate level is not increased. Neither inositol nor an agonist is required. The same effects are seen whether inositol 1,4,5‐trisphosphate is quantified by the [3H]inositol prelabeling technique or by mass assay, although mass includes a pool of inositol 1,4,5‐trisphosphate that is metabolically inactive. Thus, in a therapeutically relevant model for humans, Li+ increases inositol 1,4,5‐trisphosphate levels in brain cortex slices, as was previously seen in lower mammals at non‐rate‐limiting concentrations of inositol.
Characterization of the Alterations in Purine Nucleotide Metabolism in Hypoxanthine‐Guardne Phosphoribosyltransferase‐Deficient Rat Neuroma Cell Line
Authors: Zoref‐Shani Esther , Bromberg Yael , Brosh Sara , Sidi Yechezkel , Sperling Oded
Published: July 2010

Abstract: A rat neuroma cell line (B103 4C), deficient of hypoxanthine‐guanine phosphoribosyltransferase (HGPRT), was utilized as a model tissue in search for the biochemical basis of the Lesch‐Nyhan syndrome (LNS). The HGPRT‐deficient neurons exhibited the following properties: an almost complete absence of uptake of guanine and of hypoxanthine into intact cell nucleotides (0.92% and 0.69% of normal, respectively); a significant increase in the availability of 5′‐phosphoribosyl‐1‐pyrophosphate; a three‐ to fourfold acceleration of the rate of de novo nucleotide synthesis; a normal excretion of xanthine, but 15‐fold increase in the excretion of hypoxanthine into the culture media; a normal cellular purine nucleotide content, including the absence of 5‐amino‐4‐imidazole carboxamide nucleotides (Z‐nucleotides), but enhanced turnover of adenine nucleotides (loss of 86% of the radioactivity of the prelabeled pool in 24 h, in comparison to 73% in the normal line), and an elevated UTP content. The results suggest that, under physiological conditions, guanine salvage does not occur in the normal neurons, but that hypoxanthine salvage is of great importance in the homeostasis of the adenine nucleotide pool. The finding of the normal profile of purine nucleotides in the HGPRT‐deficient neurons indicates that the lack of hypoxanthine salvage is adequately compensated by the enhanced de novo nucleotide synthesis. These results did not furnish evidence in support of the possibility that GTP or ATP depletion, or Z‐nucleotide accumulation, occurs in HGPRT‐deficient neurons and that these are etiological factors causing the neurological abnormalities in LNS. On the other hand, the results point to the possibility that elevated hypoxanthine concentration in the brain may have an etiological role in the pathogenesis of LNS.
Inhibition of Endogenous Phosphatase in a Postsynaptic Density Fraction Allows Extensive Phosphorylation of the Major Postsynaptic Density Protein
Authors: Dosemeci Ayse , Reese Thomas S.
Published: July 2010

Abstract: The major postsynaptic density protein, proposed to be a calcium/calmodulin‐dependent protein kinase, becomes phosphorylated when a postsynaptic density preparation from rat cerebral cortex is incubated in medium containing calcium and calmodulin. Upon longer incubation, however, the level of phosphorylation declines, suggesting the presence of a phosphatase activity. When Microcystin‐LR, a phosphatase inhibitor, is included in the phosphorylation medium, the decline in phosphorylation is prevented and a higher maximal level of phosphorylation can be achieved. Under these conditions, the maximal phosphorylation of major postsynaptic density protein is accompanied by a nearly complete shift in its electrophoretic mobility from 50 kDa to 54 kDa, similar to that described for the a subunit of the soluble calcium/calmodulin‐dependent protein kinase II. Of the four major groups of serine/threonine protein phosphatases, the enzyme responsible for the dephosphorylation of major postsynaptic density protein is neither type 2C, which is insensitive to Microcystin‐LR, nor type 2B, which is calcium‐dependent. As Microcystin‐LR is much more potent than okadaic acid in inhibiting the dephosphorylation of major postsynaptic density protein, it is likely that the postsynaptic density‐associated phosphatase is a type 1. The above results indicate that the relatively low level of phosphorylation of the major postsynaptic density protein observed in preparations containing postsynaptic densities is not due to a difference between the cytoplasmic and postsynaptic density‐associated calcium/calmodulin‐dependent kinases as previously proposed, but to a phosphatase activity, presumably belonging to the type 1 group.
In Vivo Microdialysis Study of the Extracellular 3,4‐Dihydroxyphenylacetic Acid in the Rat Locus Ceruleus: Topographical and Pharmacological Aspects
Authors: Ortemann Catherine , Robert Frédéric , Renaud Bernard , Lambas‐Senas Laura
Published: July 2010

Abstract: In vivo microdialysis coupled with HPLC and electrochemical detection was used to monitor extracellular levels of 3, 4‐dihydroxyphenylacetic acid (DOPAC) of the locus ceruleus (LC) in halothane‐anesthetized rats. The identity of DOPAC was confirmed by experiments showing that the chromatographic peak was totally suppressed after inhibition of monoamine oxidase by pargyline. Histological examinations allowed to relate the quantity of DOPAC measured in the dialysates with the localization of the probe implantation site. We found that the DOPAC concentration was inversely proportional to the distance between the probe and the lateral border of the LC. Regardless of the caudorostral level of the nucleus, concentrations were maximal when the axis of the probe was 100 μM from the lateral border of the LC and decreased by 53% when this distance reached 300 μM. Activation of LC noradrenergic neurons by systemic administration of the α2‐antagonist piperoxane increased by 100% DOPAC concentrations in LC dialysates. These results suggest that the DOPAC measured by microdialysis could be considered an indicator of the functional state of LC noradrenergic neurons.