Incorporation of Glycerol and Ethanolamine into Glycerophospholipid in Rat Brain Areas During Bicuculline-Induced Convulsive Seizures

The effect of bicuculline-induced convulsive seizures on lipid metabolism has been studied in four brain areas (cerebellum, cerebral cortex, hippocampus, and brainstem) using [2-3H]glycerol and [1,2-14C]ethanolamine as radioactive lipid precursors administered simultaneously with bicuculline. Twelve minutes after the administration, the uptake of radioactivity depended both on brain area and treatment, being generally higher in convulsing rats. The uptake of glycerol was influenced to a larger extent than that of ethanolamine and increased during convulsions, but its incorporation into lipids did not. In contrast, the amount of ethanolamine incorporated into lipids increased during bicuculline-induced seizures. The difference in behavior of glycerol and of ethanolamine is also indicated by the decrease of the 3H/14C ratio of phosphatidyl-ethanolamine in various brain areas during convulsions. It is, therefore, evident that the metabolism of the two precursors is affected differently by seizures.     

Cardiac glycosides and oviposition by Danaus plexippus on Asclepias fruticosa in south-east Queensland (Australia), with notes on the effect of plant nitrogen content

Abstract. 1. Previous studies which attempt to relate oviposition by female monarchs, Danaus plexippus (L.), to the cardiac glycosides of their milkweed host plants are reviewed and found to be defective.
2. Here we followed ovipositing females in the field and recorded their oviposition behaviour on individual Asclepias fruticosa (L.) plants. The cardenolide glycoside and per cent nitrogen of each of these plants and also of a control set of plants was recorded.
3. Milkweeds accepted by females, on which egg laying took place, had significantly lower mean cardenolide levels compared with rejected plants and a control sets of plants.
4. Most oviposition ( c. 70%) took place on plants with intermediate cardenolide (CG) levels (=200–300 μg/0.1 g DW).
5. The mean CG level in A.fruticosa was 345 μg/0.1 g DW (SD = 138, n =122, range 119–719).
6. There was no relationship between monarch oviposition behaviour and plant per cent nitrogen.     

Bag-of-words representation for biomedical time series classification

Automatic analysis of biomedical time series such as electroencephalogram (EEG) and electrocardiographic (ECG) signals has attracted great interest in the community of biomedical engineering due to its important applications in medicine. In this work, a simple yet effective bag-of-words representation that is originally developed for text document analysis is extended for biomedical time series representation. In particular, similar to the bag-of-words model used in text document domain, the proposed method treats a time series as a text document and extracts local segments from the time series as words. The biomedical time series is then represented as a histogram of codewords, each entry of which is the count of a codeword appeared in the time series. Although the temporal order of the local segments is ignored, the bag-of-words representation is able to capture high-level structural information because both local and global structural information are well utilized. The performance of the bag-of-words model is validated on three datasets extracted from real EEG and ECG signals. The experimental results demonstrate that the proposed method is not only insensitive to parameters of the bag-of-words model such as local segment length and codebook size, but also robust to noise.     

Purification of a nitrate reductase kinase from Spinacea oleracea leaves, and its identification as a calmodulin-domain protein kinase

Spinach (Spinacea oleracea L.) nitrate reductase (NR) is inactivated by phosphorylation on serine-543, followed by binding of the phosphorylated enzyme to 14-3-3 proteins. We purified one of several chromatographically distinct NR_serine-543 kinases from spinach leaf extracts, and established by Edman sequencing of 80 amino acid residues that it is a calcium-dependent (calmodulin-domain) protein kinase (CDPK), with peptide sequences very similar to Arabidopsis CDPK6 (accession no. U20623; also known as CPK3). The spinach CDPK was recognized by antibodies raised against Arabidopsis CDPK. Nitrate reductase was phosphorylated at serine-543 by bacterially expressed His-tagged CDPK6, and the phosphorylated NR was inhibited by 14-3-3 proteins. However, the bacterially expressed CDPK6 had a specific activity approx. 200-fold lower than that of the purified spinach enzyme. The physiological control of NR by CDPK is discussed, and the regulatory properties of the purified CDPK are considered with reference to current models for reversible intramolecular binding of the calmodulin-like domain to the autoinhibitory junction of CDPKs. Received: 12 February 1998 / Accepted: 28 May 1998     

Lipophosphoglycan Antigen Shedding By Leishmania Donovani

ABSTRACT. The biochemical characterizations of lipophosphoglycans from various Leishmania species reported by other workers may or may not contain several types of lipophosphoglycan molecules. This is the first report in which a specific lipophosphoglycan has been defined by both its antigenie and electrophoretic properties. Furthermore, a purification procedure for this specific lipophosphoglycan is described and some biochemical characterizations are presented. Phospholipase C and the so-called phosphatidylinositol-specific phospholipase C of Bacillus cereus convert the amphipathic form of the lipophosphoglycan antigen to the hydrophilic form. Under equivalent incubation conditions, other phospholipases tested were not effective in conversion of the amphipathic to the hydrophilic form. Since the amphipathic form is present in conditioned media, antigen shedding cannot be explained by phospholipase C digestion of the amphipathic form, which would result in the release of only the hydrophilic form into the medium. Both the pellet and the supernatant fractions of conditioned media contained both forms of the antigen and did not differ in the relative amounts of the two. This observation rules out membrane blebbing as the major mechanism for the release of the amphipathic form.     

Knockout of Nr2e3 prevents rod photoreceptor differentiation and leads to selective L-/M-cone photoreceptor degeneration in zebrafish

Mutations in the photoreceptor cell-specific nuclear receptor gene Nr2e3 increased the number of S-cone photoreceptors in human and murine retinas and led to retinal degeneration that involved photoreceptor and non-photoreceptor cells. The mechanisms underlying these complex phenotypes remain unclear. In the hope of understanding the precise role of Nr2e3 in photoreceptor cell fate determination and differentiation, we generated a line of Nr2e3 knockout zebrafish using CRISPR technology. In these Nr2e3-null animals, rod precursors undergo terminal mitoses but fail to differentiate as rods. Rod-specific genes are not expressed and the outer segment (OS) fails to form. Formation and differentiation of cone photoreceptors is normal. Specifically, there is no increase in the number of UV-cone or S-cone photoreceptors. Laminated retinal structure is maintained. After normal development, L-/M-cones selectively degenerate, with progressive shortening of OS that starts at age 1 month. The amount of cone phototransduction proteins is concomitantly reduced, whereas UV- and S-cones have normal OS lengths even at age 10 months. In vitro studies show Nr2e3 synergizes with Crx and Nrl to enhance rhodopsin gene expression. Nr2e3 does not affect cone opsin expression. Our results extend the knowledge of Nr2e3's roles and have specific implications for the interpretation of the phenotypes observed in human and murine retinas. Furthermore, our model may offer new opportunities in finding treatments for enhanced S-cone syndrome (ESCS) and other retinal degenerative diseases.     

The homologous tryptophan critical for cytochrome c peroxidase function is not essential for ascorbate peroxidase activity

The crystal structures of ascorbate peroxidase (APX) and cytochrome c peroxidase (CCP) show that the active site structures are nearly identical. Both enzymes contain a His-Asp-Trp catalytic triad in the proximal pocket. The proximal Asp residue hydrogen bonds with both the His proximal heme ligand and the indole ring nitrogen of the proximal Trp. The Trp is stacked parallel to and in contact with the proximal His ligand. This Trp is known to be the site of free radical formation in CCP compound I and also is essential for activity. However, APX forms a porphyrin radical and not a Trp-centered radical, even though the His-Asp-Trp triad structure is the same in both peroxidases. We found that conversion of the proximal Trp to Phe has no effect on APX enzyme activity and that the mutant crystal structure shows that changes in the structure are confined to the site of mutation. This indicates that the paths of electron transfer in CCP and APX are distinctly different. The Trp-to-Phe mutant does alter the stability of the APX compound I porphyrin radical, by a factor of two. Electrostatic calculations and modeling studies show that a potassium cation located about 8?Å from the proximal Trp in APX, but absent in CCP, makes a significant contribution to the stability of a cation Trp radical. This underscores the importance of long-range electrostatic effects in enzyme catalyzed reactions.     

Précisions Temporelles sur les Processus de la Stomatogenèse de Tetrahymena paravorax analysés en Microscopie Photonique

Resume. Une analyse des séquences morphogénétiques du CiliéTetrahymena paravorax montre que: (A) La durée de la stomatogenèse de bipartition des formes microstomes en croissance exponentielle représente 45% du temps de génération (stade 1—20%; stade 2—3%; stade 3—3%; stade 4—5%; stade 5—5%; stade 6—9%). (B) La division cytoplasmique est inégale (les proters sont plus petits que les opisthes); la différence de taille initiale entre les 2 produits de fission est probablement compensée par une prolongation de la période de croissance chez le proter. (C) Le pourcentage maximum de réorganisation buccale microstome → macrostome pour les populations asynchrones atteint –? 70% au bout de 210 mn d'incubation dans la stomatine. (D) L'initiation de la stomatogenèse de remplacement oral est connectée avec la fin d'une période dont la durée minimale est approximativement celle du stade 0 du cycle normal d'interdivision des microstomes; cette initiation est retardée chez les microstomes exposés à la stomatine dès le début du cycle cellulaire. (E) Le primordium buccal de division peut se résorber en présence de stomatine et la stomatogenèse antérieure peut commencer avant que ne soit terminée cette résorption; la résorption n'est plus induite au-delà d'un point du stade 5 qui précède le début de la constriction du corps cellulaire. SYNOPSIS. An analysis of the morphogenetic sequences in the ciliate Tetrahymena paravorax has shown that: (A) The duration of predivision stomatogenesis in exponentially growing microstomes occupies 45% of the generation time (stage 1—20%; stage 2—3%; stage 3—3%; stage 4—5%; stage 5—5%; stage 6—9%). (B) Cytoplasmic division is unequal (the proters are smaller than opisthes); the initial size difference between the 2 fission products is presumably compensated by an increased growth period in the proter. (C) The maximum percentage of microstome-to-macrostome oral reorganization is –? 70% in asynchronous populations, 210 min after suspension in stomatin. (D) Initiation of oral replacement stomatogenesis is associated with the end of a period which has a minimum duration nearly equal to that of stage 0 characteristic of the normal inter-division cycle of the microstomes; this initiation is delayed if exposure of microstomes to stomatin is begun at the onset of the cell cycle. (E) The buccal primordium formed in division can be resorbed in presence of stomatin and anterior stomatogenesis can start before the resorption is completed: this resorption is not induced if the cells have progressed beyond a point which precedes the beginning of the cell furrowing (stage 5).