A micro-method for the assay of polyadenylate-containing ribonucleic acid by gel electrophoresis.

A micro-method for the determination of the electrophoretic profile of the various poly(A)-containing RNA species in a RNA sample was developed. The method is simple to carry out and allows for great reproducibility. It combines the resolving power of electrophoresis in agarose with the specificity of binding of poly(A) to poly(U)-containing glass-fibre filters. It consists of the following steps. (1) The molecules in an RNA sample are first separated according to their molecular weight by electrophoresis in agarose, at low ionic strength. 2. The molecules thus seperated are then submitted to a second electrophoresis in 'binding buffer' in a direction perpendicular to the first one. In the course of this electrophoresis the poly(A)-containing RNA species are seperated from other RNA species as they bind to a poly(U)-containing glass-fibre filter which is placed across the electrophoresis path. The method was used to determine the electrophoretic profile of Rhynchosciara salivary-gland messenger RNA. It was found that the population of messenger RNA in the gland is dominated by forms moving as 18 and 158 S peaks, but there is also polydisperse RNA with slower mobility.     

Neurotrophic Effects of Extracellular Guanosine

Central nervous system (CNS) astrocytes release guanosine extracellularly, that exerts trophic effects. In CNS, extracellular guanosine (GUO) stimulates mitosis, synthesis of trophic factors, and cell differentiation, including neuritogenesis, is neuroprotective, and reduces apoptosis due to several stimuli. Specific receptor-like binding sites for eGUO in the nervous system may mediate its effects through both MAP kinase and PI3-kinase signalling pathways. Extracellular guanine (eGUA) also exerts several effects; the trophic effects of eGUO are likely regulated by conversion of eGUO to eGUA by a membrane located purine nucleoside phosphorylase (ecto-PNP) and by conversion of eGUA to xanthine by guanine deaminase.     

Midtrimester pregnancy interruption and the placental progesterone levels

Placental progesterone contents were studied in 10 patients therapeutically aborted at midtrimester by intraamniotic infusions of hypertonic saline, and from 14 patients (12–20 weeks) aborted by intraamniotic instillation of prostaglandin F_2∝. The mean S.E. of the progesterone was 1.99± 0.07 ug/g. placental tissue in the first group, while with prostaglandin abortion the placental progesterone was 1.45± 0.09 ug/g. tissue, which is significantly lower than the results in the first group (P=0.001). The possible mechanism of action of prostaglandin as an effective abortifacient is discussed.     

Resistance of the AF-28 sorghum variety to Contarinia sorghicola

The variety AF-28 of Sorghum bicolor Pers and the hybrid R-1090 were compared in relation to the sorghum midge, Contarinia sorghicola (Coquillet), (Diptera: Cecidomyidae) in two counties of the State of São Paulo. They were compared in seven successive monthly trials beginning in September 1972. Averages of 252 and 137 adult flies emerged per head of the hybrid R-1090 in Campinas and Jaboticabal respectively, while 2.9 and 0.8 emerged from AF-28. The damage was graded with a visual scale from 0 to 4. Average damage grades of 3.5 and 3.3 were attributed to the hybrid R-1090 in Campinas and Jaboticabal respectively, while the variety AF-28 had averages of 0.5 and 0.3, respectively.

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Résumé La variété AF-28 et l'hybride R-1090 de Sorghum bicolor Pers. ont été comparés à différentes époques de l'année et en deux localités différentes de l'Etat de Sâo Paulo, Brésil.On n'a pas constaté d'interaction variété x époque ou variété x lieu.AF-28 a prouvé sa résistance à toutes les époques, aussi bien dans un lieu que dans l'autre; l'hybride par contre s'est montré grandement susceptible.Des moyennes de 252,2 et 136,7 mouches par panicule ont été observées pour l'hybride R-1090 à Campinas et à Jaboticabal contre 2,9 et 0,8 seulement par panicule de AF-28. Le préjudice a été estimé visuellement selon une échelle de 0 à 4. Des préjudices d'une valeur moyenne de 3,5 et 3,3 ont été attribués à l'hybride R-1090 à Campinas et à Jaboticabal respectivement, alors que simultanément la variété AF-28 ne subissait que des préjudices évalués à 0,5 et 0,3 respectivement.Quatre espèces de microhyménoptères parasites de Contarinia sorghicola ont été récoltées au cours de ce travail: Eupelmus popa et trois espèces différentes du genre Tetrastichus.

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Part of a thesis presented to the School of Sciences of Bauru for fulfilment of the requirement for the degree of Doctor of Science. This research was financed by the Project 72/1471 of the agreement UNIÃO-ESTADO-FAPESP and by CNPq Project 12910/73.     

Chromatin modifiers and recombination factors promote a telomere fold-back structure,that is lost during replicative senescence

Telomeres have the ability to adopt a lariat conformation and hence, engage in long and short distance intra-chromosome interactions. Budding yeast telomeres were proposed to fold back into subtelomeric regions, but a robust assay to quantitatively characterize this structure has been lacking. Therefore, it is not well understood how the interactions between telomeres and non-telomeric regions are established and regulated. We employ a telomere chromosome conformation capture (Telo-3C) approach to directly analyze telomere folding and its maintenance in S. cerevisiae. We identify the histone modifiers Sir2, Sin3 and Set2 as critical regulators for telomere folding, which suggests that a distinct telomeric chromatin environment is a major requirement for the folding of yeast telomeres. We demonstrate that telomeres are not folded when cells enter replicative senescence, which occurs independently of short telomere length. Indeed, Sir2, Sin3 and Set2 protein levels are decreased during senescence and their absence may thereby prevent telomere folding. Additionally, we show that the homologous recombination machinery, including the Rad51 and Rad52 proteins, as well as the checkpoint component Rad53 are essential for establishing the telomere fold-back structure. This study outlines a method to interrogate telomere-subtelomere interactions at a single unmodified yeast telomere. Using this method, we provide insights into how the spatial arrangement of the chromosome end structure is established and demonstrate that telomere folding is compromised throughout replicative senescence.     

Impact of brood parasitism and predation on nest survival of the fan-tailed gerygone in New Caledonia

Predation and brood parasitism are common reasons for nesting failure in passerine species and the additive impact by invasive species is a major conservation concern, particularly on tropical islands. Recognising the relative contribution of the different components of nesting failure rates is important to understand co-evolutionary interactions within brood parasite–host systems. In the remote archipelago of New Caledonia, the fan-tailed gerygone Gerygone flavolateralis is the exclusive host of the brood-parasitic shining bronze-cuckoo Chalcites lucidus. Additionally, invasive rodents also possibly have an impact on breeding success. To estimate the impact of potential nest predators, we 1) video monitored nests to identify predators, 2) estimated the probability of predation based on nest visibility and predator abundance and 3) tested the possibility that the location of experimental nests and lack of odour cues decrease the predation by rodents. In addition, we estimated nest survival rates using data collected in different habitats over the course of eight breeding seasons. Nesting success of fan-tailed gerygones was relatively low and predation was the main cause of nesting failure. We recorded mainly predation by native birds, including the shining bronze-cuckoo, whereas predation by rats was rare. In open habitats predation by cuckoos was much lower than predation by other avian predators. Neither predator activity around nests nor nest visibility influenced the probability of predation. Experimental nests in more accessible locations and containing an odorous bait were more exposed to rodent predation. Apparently, the fan-tailed gerygone has either never been specifically vulnerable to predation by rats or has developed anti-predator adaptations.