Characterization of a novel plant poly(A) polymerase

We have purified and characterized poly(A) polymerases (PAPs) from Pisum sativum, Brassica juncea, and Zea mays. Through chromatography on DEAE-Sepharose and heparin-Sepharose, these PAPs copurified as a single enzyme along with RNPs that could provide RNA substrates for the enzyme. More extensive purification by chromatography on MonoQ resulted in the resolution of the PAPs into as many as three fractions. One of these (PAP-I) contained a 43-kDa polypeptide immunologically related to the yeast PAP, and two others (PAP-II and PAP-III) contained RNAs that could serve as substrates for polyadenylation. These fractions by themselves possessed little PAP activity, but mixtures containing combinations of these displayed substantial activity. Similar PAP factors (PAP-I and PAP-III) were identified after fractionation of extracts prepared from Brassica juncea and Zea mays. The factors from one plant were completely interchangeable with those from different plants. We conclude that the poly(A) polymerases present in vegetative plant tissues consist of more than one component. In this respect, they are substantially different from other reported plant, mammalian, and yeast PAPs.     

An evaluation of ‘rapid review’ as a method of quality control of cervical smears using the AxioHOME microscope

An evaluation of ‘rapid review’ as a method of quality control of cervical smears using the AxioHOME miscroscope One method of quality control which has recently been recommended by professional bodies in the UK is the ‘rapid review’ method. This involves the microscopic 30 s review of all negative cervical smears with the intention of flagging potential missed abnormalities. Although it has been suggested that rapid review is better than 10% random rescreening of negative smears, the efficiency and efficacy of this method of quality control have not been thoroughly evaluated. We have used the AxioHOME system, which can record the area of a slide covered and the screening time, to investigate slide coverage during rapid review quality control, as performed by 15 cytoscreeners and MLSOs reviewing a test set of 22 slides each. The test set comprised 18 negative slides, three positive slides, and one unsatisfactory slide. We have recorded two distinct methods of rapid review in use amongst cytotechnologists, the step method and the whole slide method. The data show that rapid review takes longer on average than the recommended 30 s, the mean screening times being 76 s and 82 s for the step and whole slide methods, respectively. Abnormal smears were missed on three of 15 occasions by the step method (sensitivity 80%, positive predictive value 85%), and on seven of 30 occasions by the whole slide method (sensitivity 76.6%, positive predictive value 45%). However, the 95% confidence intervals were wide (57.7–90.7% for the step method, and 51.9–95.7% for the whole slide method). Analysis of scanning tracks and screening rates shows significant flaws in the methodology of rapid review. Abnormal cells were not identified, although dyskaryotic cells were included in the scanning track on nine occasions, seven using the whole slide method and two using the step method. On one occasion (using the step method) abnormal cells were not identified because they were not included in the scanning track. Further research is in progress to determine optimal methods of rapid review, and whether the rapid review technique is as effective as automated screening systems for quality assurance in cytology. Evaluation de la technique de ‘Relecture Rapide’ comme méthode de contrôle de qualité des frottis cervico-utérins, à l'aide du microscope AxioHOME Une des méthodes de contrôle de qualité récemment recommandée par le corps professionnel du Royaume Uni est la méthode dite de ‘relecture rapide’. Cette méthode consiste en une deuxième lecture d'une durée de trente secondes de tous les frottis cervicaux négatifs et dont l'objectif est de détecter les anomalies ayant pu échapper au premier examen. Bien qu'il ait été suggéré que cette méthode de relecture rapide soit meilleure que la relecture de 10% des frottis négatifs tirés au sort, le rendement et l'efficacité de cette méthode de contrôle de qualité n'ont pas été évalués complètement. Nous avons utilisé le système AxioHOME capable d'enregistrer la plage de la lame qui a été explorée ainsi que le temps de lecture afin d'étudier la surface explorée au cours de cette relecture rapide telle qu'elle a pu être pratiquée par quinze cytotechniciens et MLSOs, chacun ayant relu une série test de vingt deux lames. Cette série test comprenait dix huit lames négatives, trois lames positives et un frottis non satisfaisant. Nous avons noté que les cytotechniciens utilisaient deux méthodes de relecture rapide différentes, la méthode ‘pas à pas’ et la méthode ‘globale’. Les données montrent que la relecture rapide prend, en moyenne, un temps supérieur aux trente secondes recommandées, la moyenne des temps de lecture étant de 76 secondes et de 82 secondes respectivement pour la méthode ‘pas à pas’ et la méthode ‘globale’. Les anomalies n'ont pas été détectées dans trois cas sur quinze par la méthode ‘pas à pas’ (sensibilité 80%, valeur prédictive positive 85%), et dans 7 cas sur 30 par la méthode globale (sensibilité 76,6%, valeur prédictive positive 45%). Toutefois, l'intervalle de confiance à 95% est important (57,7%-90,7% pour la méthode ‘pas à pas’ et 51,9%-95,7% pour la méthode globale). L'analyse des surfaces balayées et des taux de détection montre des points faibles significatifs de cette méthodologie de relecture rapide. Dans 9 cas, les anomalies n'ont pas été identifiées alors que des cellules dyskaryotiques étaient présentes dans les plages balayées au cours de la relecture (sept utilisant la méthode globale et deux utilisant la méthode ‘pas à pas’). Dans un cas (avec la méthode ‘pas à pas’) les cellules anormales n'ont pas été identifiées parce qu'elles étaient absentes des plages de relecture. Des études sont en cours afin de déterminer quelles sont les méthodes optimales de relecture rapide et si ces techniques de relecture rapide sont aussi efficaces que les systèmes de lecture automatisée pour l'assurance de qualité en cytologie cervico-utérine. ‘Rapid Review’ als Methode der Qualitätskontrolle gynäkologischer Abstriche, Überprüfung mit dem AxioHOME-Mikroskop Die empfohlene ‘Rapid Review’ Kontrolle aller negativen Abstriche in nur 30 Sekunden anstelle des Nachscreenens von 10% der Präparate ist in ihrer Zuverlässigkeit bislang nicht überprüft worden. Mit Hilfe des AxioHOME-Mikroskops ist es möglich sowohl die ausgewertete Fläche, als auch die erforderliche Zeit zu erfassen. 15 Auswerter prüften mit der Methode jeweils einen Testsatz von 22 Präparaten. Er enhielt 18 negative, 3 positive und 1 nichtauswertbaren Abstrich. Getestet wurden zwei verschiedene Vorgehensweisen: die schrittweise und die das ganze Präparat erfassende. Beide erfordern mehr Zeit als 30 Sekunden; der mittlere Zeitaufwand betrug für die Schrittmethode 76 und für die Ganzheitsmethode 82 Sekunden. Die Schrittmethode verfehlte 3/15 Anomalien (Sensitivität 80%, positiver prädiktiver Wert 85%), die Ganzheitsmethode 7/30 (Sensitivität 76,6%, positiver prädiktiver Wert 45%). Der 95% Konfi denzbereich reichte für die Schrittmethode von 57,5–90,7% und für die Ganzheitsmethode von 51,9–95,7%). Die Analyse deckt wesentliche Schwachstellen des Rapid Review-Verfahrens auf. In 9 Fällen lagen nicht erkannte Zellatypien in den kontroll ierten Bahnen (7 bei des Schrittmethode, 2 bei der Ganzheitsmethode). Einmal lagen sie bei der Schrittmethode ausserhalb der geprüften Bahnen. Weitere Studien werden prüfen ab das Rapid Review-Verfahren automatisierten Systemen vergleichbar ist.     

Twin-screw extrusion of ‘Pesta’-encapsulated biocontrol agents

World Journal of Microbiology and Biotechnology -     

Hyperglycemic Damage to Mitochondrial Membranes During Cerebral Ischemia: Amelioration by Platelet-Activating Factor Antagonist BN 50739

Abstract: The Pulsinelli-Brierley four-vessel occlusion model was used to study the consequences of hyperglycemic ischemia and reperfusion. Rats were subjected to either 30 min of normo- or hyperglycemic ischemia or 30 min of normo- or hyperglycemic ischemia followed by 60 min of reperfusion. In some animals, 2 mg/kg BN 50739, a platelet-activating factor receptor antagonist, was administered intraarterially either before or after the ischemic insult. The changes in mitochondrial membrane free fatty acid levels, phosphatidylcholine fatty acyl composition, and thiobarbituric acid-reactive material (TBAR) content plus the mitochondrial respiratory control ratio (RCR) were monitored. When the platelet-activating factor antagonist was present during normoglycemia, (a) the mitochondrial free fatty acid release both during and after ischemia was slowed, (b) reacylation of phosphatidylcholine following ischemia was promoted, and (c) TBAR accumulation during and following ischemia was decreased. The detrimental effects of hyperglycemia were muted when BN 50739 was present during ischemia. The RCR was preserved and phosphatidylcholine hydrolysis during ischemia was decreased. TBAR levels were consistently higher in hyperglycemic brain mitochondria both during and after ischemia. The RCR correlated directly with mitochondrial phosphatidylcholine polyunsaturated fatty acid content during ischemia and reperfusion. BN 50739 protection of mitochondrial membranes in brain may be influenced by tissue pH.     

甘蔗细平象的研究

甘蔗细平象Trockorhopalus humeralis Chevrolat是云南甘蔗上的一种毁灭性地下害虫,以幼虫和成虫在地下蔗头内为害,为害期8-10个月。据1989年调查,受害蔗每亩损失500-3000kg,严重的无收。此虫1年发生1代,以成虫在蔗头内越冬,有喜湿性,不能飞翔,主要通过沟河流水传播。在河川坝地,沙壤土中虫口较多;宿根年限越长的甘蔗受害越重。建议蔗稻轮作;缩短甘蔗宿根年限;早春翻挖有虫蔗蔸烧毁;结合新植蔗下种,宿根蔗松蔸培土施用甲基异柳磷或铁灭克等颗粒杀虫剂进行防治。     

Population suppression for control of the blowfly Lucilia sericata and sheep blowfly strike

Abstract.

• 1 The control of ovine myiasis by suppression of populations of the blowfly Lucifia sericata was investigated experimentally on three farms in the south-west of England in 1992 and 1993.
• 2 In blind trials, sheep on one farm (control) were given two doses of placebo, on a second two doses of the larvicide cyromazine (Vetrazin®, CibaGeigy), and on a third cyromazine and a subsequent dose of placebo.
• 3 The first treatment was given shortly before the predicted spring emergence of L.sericata and the second shortly before the predicted emergence of the second generation. Previous simulation analysis had identified strategic early-season treatment as the optimum for blowfly population suppression.
• 4 On both treatment farms significantly smaller L.sericata populations were recorded throughout 1992 and the incidence of strike was significantly lower than on the control farm. The results show that appropriate early-season timing of sheep treatment can suppress populations of L.sericata and could be used by farmers to reduce the incidence of blowfly strike.
• 5 The results suggest, however, that the effectiveness of population suppression and strike incidence may have been influenced by immigration into the control areas and by adverse weather, the latter changing the susceptibility of sheep to strike and resulting in rising strike incidence even when L.sericata population densities were low. In practice, therefore, blowfly population suppression should be employed as a component of an integrated strike management programme.

     

Interspecific host discrimination and competition in Apoanagyrus (Epidinocarsis) lopezi and A.(E.)diversicornis, parasitoids of the cassava mealybug Phenacoccus manihoti

Abstract.

• 1 Choice experiments on interspecific host discrimination in A.lopezi and A. diversicornis were carried out on discs of cassava leaf containing four hosts (P.manihoti) that had been parasitized by the other species and four unparasitized hosts.
• 2 A.lopezi accepted both host types equally for oviposition, whereas A.diversicornis accepted fewer hosts that had been parasitized by A.lopezi than unparasitized ones. A.diversicornis is therefore capable of interspecific host discrimination, but such a capability was not demonstrated for A.lopezi.
• 3 Survival probability in singly parasitized hosts was 0.85 for both parasitoid species. When the time interval between ovipositions was 2 h or less, survival in multiparasitized hosts was 0.68 for A.lopezi and 0.17 for A.diversicornis, irrespective of priority. Increasing A.lopezi priority to 24±2h did not increase A.lopezi survival. A.diversicornis survival, however, increased to 0.43 when A.diversicornis was given 24 ± 2 h priority. A.diversicornis eggs took 19 h longer than A.lopezi eggs to hatch. This could explain the difference in competitive abilities in multiparasitized hosts.
• 4 The observed difference in host selection behaviour between A.lopezi and A.diversicornis is in accordance with the different benefits of multiparasitism: A.lopezi gains more than A.diversicornis because of its superior within-host competitive abilities.
• 5 Neither species avoided multiparasitism completely. The low survival probability of A.diversicornis in multiparasitized hosts may partly explain its failure to establish when introduced into Africa as part of a biological control programme of P.manihoti.

     

Recent advances in plant cell cultures in bioreactors

Two key issues in the application of plant-cell-culture technology to the production of valuable secondary metabolites are reviewed: the selection of cell lines with suitable genetic, biochemical and physiological characteristics; and the optimization of bioreactor environments. Although great progress has been made in recent years in the design, selection and optimization of bioreactor hardware, optimization of environmental factors such as medium components, light irradiation and O₂ supply needs detailed investigations for each case. With a better understanding of plant cell metabolism and physiology, further developments in cultivation processes, such as process integration and on-line monitoring and control, can be expected in the near future.J.-J. Zhong and J.-T. Yu are with the Research Institute of Biochemical Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, China T. Yoshida is with the International Center of Cooperative Research in Biotechnology (ICBiotech), Faculty of Engineering, Osaka University, Suita, Osaka 565, Japan.     

Schistosoma mansoni: differences in acetylcholine, dopamine, and serotonin control of circular and longitudinal parasite muscles

The physiological and pharmacological properties of circular and longitudinal somatic musculature in adult male Schistosoma mansoni were compared using cut muscle sections. Carbachol reduced tone in both circular and longitudinal muscle, but was without effect on circular muscle bathed in high Mg2+, indicating that cholinergic receptors were not associated with circular muscle membrane. 5-Hydroxytryptamine (5-HT) induced rhythmic contractile activity in both sets of muscle. It decreased muscle tone in circular muscle but increased the tone of longitudinal muscle. Metergoline blocked 5-HT effects on both sets of muscle. 5-HT continued to be effective on both sets of muscle bathed in high-Mg2+ medium, indicating that serotonergic receptors were present on both circular and longitudinal muscle membranes. Dopamine decreased both circular and longitudinal muscle tone. Its effects on circular muscle were still present after exposure to high Mg2+, but its effects on the longitudinal muscle were significantly reduced, leading to the conclusion that dopaminergic sites were probably associated with circular muscle membrane but not that of longitudinal muscle. Also, spiroperidol blocked stimulus responsiveness of the circular muscle but not that of the longitudinal muscle. From these studies it appears that there are significant physiological and pharmacological differences between circular and longitudinal muscles in the adult male schistosome.