Large-scale physical mapping within the region 22q12.3-13.1 in meningioma

The lack of physical mapping data strongly restricts the analysis of the meningioma chromosomal region that was assigned to the bands 22q12.3-qter. Recently, we reported a new marker D22S16 for chromosome 22 that was assigned to the region 22q13-qter by in situ hybridization. Utilizing somatic cell hybrids we now sublocalized the marker D22S16 within the band region 22q12-13.1, thus placing it in the vicinity of the gene for the platelet derived growth factor (PDGFB). A physical map was established for the regions surrounding the PDGFB gene and the D22S16 marker. By means of pulsed-field gel electrophoresis (PFGE) D22S16 and PDGFB were found to be physically linked within 900 kb. We also identified two CpG clusters bordering the PDGFB gene. For the enzyme NotI, a variation of the PDGFB restriction pattern was found between different individuals. PFGE analysis of the two loci (PDFGB and D22S16) failed to identify major rearrangements in meningioma.     

Acquired thermotolerance and heat shock in the extremely thermophilic archaebacterium Sulfolobus sp. strain B12.

The extreme thermophile Sulfolobus sp. strain B12 exhibits an acquired thermotolerance response. Thus, survival of cells from a 70 degrees C culture at the lethal temperature of 92 degrees C was enhanced by as much as 6 orders of magnitude over a 2-h period if the culture was preheated to 88 degrees C for 60 min or longer before being exposed to the lethal temperature. In eubacteria and eucaryotes, acquired thermotolerance correlates with the induced synthesis of a dozen or so proteins known as heat shock proteins. In this Sulfolobus species, it correlates with the preferential synthesis of primarily one major protein (55 kilodaltons) and, to a much lesser extent, two minor proteins (28 and 35 kilodaltons). Since the synthesis of all other proteins was radically reduced and these proteins were apparently not degraded or exported, their relative abundance within the cell increased during the time the cells were becoming thermotolerant. They could not yet be related to known heat shock proteins. In immunoassays, they were not cross-reactive with antibodies against heat shock proteins from Escherichia coli (DnaK and GroE), which are highly conserved between eubacteria and eucaryotes. However, it appears that if acquired thermotolerance depends on the synthesis of protective proteins, then in this extremely thermophilic archaebacterium it depends primarily on one protein.     

Alpha-globin gene markers identify genetic differences between Australian aborigines and Melanesians.

Australian aborigines exhibit a number of alpha-globin cluster rearrangements involving both alpha- and zeta-globin genes. alpha+-Thalassemia (-alpha/) in this population is heterogeneous and includes the 3.7 types I, II, and III gene deletions. The alpha alpha alpha/ and zeta zeta zeta/ rearrangements are each found in association with two haplotypes, indicating origins from at least two separate DNA crossover events. Differences in alpha-globin cluster rearrangements and in haplotypes between Australian aborigines, Papua New Guinea highlanders and island Melanesians, are consistent with multiple colonizing events into Australia.     

Messenger RNA is translated when associated with the cytoskeletal framework in normal and VSV-infected HeLa cells

When the cytoskeletal framework is prepared from suspension-grown HeLa by extraction with nonionic detergent, all the polyribosomes are associated with the framework while 80% of tRNA and the major portion of monoribosomes as well as 75% of the cell proteins are found in the soluble fraction. The mRNA of polyribosomes is bound to the cytoskeleton and these molecules remain attached even after polyribosomes are disassembled in vivo prior to extraction. Although all actively translating message molecules are attached to the framework, about one quarter of the poly(A)+ mRNA is free of the framework. The binding of message to the skeleton may be obligatory for translation. Upon infection with VSV, all the viral polyribosomes but not all the viral messages of the infected cell are associated with the cytoskeletal framework. Pulse-chase labeling shows that VSV messages initially associate with the framework and then later detach and cease translation. The mRNA for the viral glycoprotein (G), known to translate only on ribosomes bound to endoplasmic reticulum, is also retained by the detergent-extracted structure. It appears that the protein substructure of the endoplasmic reticulum which binds polyribosomes is a component of the cytoskeletal framework.     

Effects of fenvalerate and azinphosmethyl on two-spotted spider mite and phytoseiid mites

The pyrethroid fenvalerate showed significantly faster activity against adult two-spotted spider mite Tetranychus urticae Koch c.f. azinphosmethyl using broad bean leaf discs sprayed in a Potter tower. LC₅₀s for fenvalerate were similar at 24 and 48 hr (0.056 and 0.051g AI/1) while LC₅₀s for azinphosmethyl were significantly different at 24 and 48 hr (0.72 and 0.38g AI/1, respectively). Mortality was partitioned to run-off and direct mortality. Fenvalerate showed an increasing contribution to mortality by run-off with increasing concentration. Increasing concentrations of azinphosmethyl had no effect on the proportion of T. urticae running off the discs. Fenvalerate inhibited egg production c.f. azinphosmethyl (60% and 20% inhibition respectively c.f. control after 24hr). The effect was not permanent. Carbaryl showed no acaricidal or inhibitory effects at 1g AI/1. T. urticae detected fenvalerate residues as reflected by choice of oviposition sites on untreated halves of leaf discs c.f. treated halves. Azinphosmethyl had no effect on oviposition preference. Phytoseiid mites were highly sensitive to fenvalerate residues. Predators moved off the test arena into sticky barriers after feeding on fenvalerate-treated eggs or walking on glass slides treated at 0.00005g AI/1.

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Résumé Les insecticides pyrethroïdes ont été utilisés pour lutter contre les pullulations d'araignées rouges. Cette note examine les réponses de Tetranychus urticae Koch et des prédateurs phytoseiidés résistants aux organo-phosphorés, Amblyseius fallacis Garman et Typhlodromus occidentalis Nesbitt, au fenvalerate (pyrethroïde) et à l'azinphosmethyl (organophosphate). Quelques essais avec du carbaryl sont indiqués.Une femelle adulte de T. urticae est placée sur une rondelle de feuille de Phaseolus vulgaris L. pulvérisée dans une tour de Potter.Les résultats sur la mortalité en fonction de la dose obtenus montrent une activité plus rapide du fenvalerate que de l'azinphosmethyl. Les DL₅₀ du fenvalerate (0,056 et 0,051g AI/I) sont les mêmes à 24 et 48 h, tandis que l'azinphosmethyl montre une activité plus lente (DL₅₀ de 0,72 et 0,38g AI/I à 24 et 48 h). La mortalité se partage entre la sortie de la rondelle et la mortalité in situ.Le fenvalerate provoque une plus forte répulsion que l'azinphosmethyl. Contrairement à l'azinphosmethyl le fenvalerate inhibe la production d'oeufs 60% et 20% d'inhibition à DL50 au bout de 24 h par rapport au témoin. Cette inhibition n'est pas permanente. Le carbaryl n'a pas d'effets inhibiteur ou acaricide à 1g AI/l.Les femelles adultes de T. urticae décèlent les résidus de fenvalerate sur les rondelles et pondent leurs oeufs sur les moitiés non traitées ou traitées à l'azinphosmethyl.Les Phytoseiides sont très sensibles aux résidus de fenvalerate. Après consommation d'oeufs traités, A. fallacis est incapable d'éviter des bandes gluantes. T. occidentalis décèle des traitements à 0,00005g AI/l en quittant les lames traitées par les bandes gluantes.

     

Messenger RNA in HeLa cells: kinetics of formation and decay

The polyadenylic acid-containing messenger RNA fraction of HeLa cells was measured by its affinity for oligedeoxythymidylate cellulose. Both the kinetics of initial labeling and the decay after a brief pulse of incorporation were examined.The kinetics of decay are complex, but can be approximated by assuming two populations; a short-lived species with a half-life of seven hours and a long-lived component with a half-life of 24 hours. It is estimated that the short-lived material comprises 33% of total cellular mRNA, while the relatively stable species amounts to 67% of the steady-state mRNA content.The two mRNA components with different decay times were observed simultaneously in the same cell population by measuring decay of 24-hour old mRNA labeled with ¹⁴C and RNA briefly labeled with ³H. The old mRNA had only a 24-hour decay component, while the new mRNA was biphasic. The decay of old and new mRNA was also observed after RNA synthesis was inhibited with actinomycin. Again, old mRNA decayed more slowly than recently labeled material. However, both decay times are significantly shorter in the presence of actinomycin and correspond to half-lives of approximately 4 and 12 hours.There is a small but significant difference in sedimentation distribution of new and old mRNA, the old mRNA sedimenting more slowly than new material, suggesting that the more stable species has a lower average molecular weight.The steady-state content of mRNA in HeLa cells amounts to 5.5% of the ribosomal RNA, or more than twice the amount of messenger RNA estimated to be on hemoglobin-synthesizing polyribosomes.     

Conserved and divergent aspects of Robo receptor signaling and regulation between Drosophila Robo1 and C. elegans SAX-3

The evolutionarily conserved Roundabout (Robo) family of axon guidance receptors control midline crossing of axons in response to the midline repellant ligand Slit in bilaterian animals including insects, nematodes, and vertebrates. Despite this strong evolutionary conservation, it is unclear whether the signaling mechanism(s) downstream of Robo receptors are similarly conserved. To directly compare midline repulsive signaling in Robo family members from different species, here we use a transgenic approach to express the Robo family receptor SAX-3 from the nematode Caenorhabditis elegans in neurons of the fruit fly, Drosophila melanogaster. We examine SAX-3’s ability to repel Drosophila axons from the Slit-expressing midline in gain of function assays, and test SAX-3’s ability to substitute for Drosophila Robo1 during fly embryonic development in genetic rescue experiments. We show that C. elegans SAX-3 is properly translated and localized to neuronal axons when expressed in the Drosophila embryonic CNS, and that SAX-3 can signal midline repulsion in Drosophila embryonic neurons, although not as efficiently as Drosophila Robo1. Using a series of Robo1/SAX-3 chimeras, we show that the SAX-3 cytoplasmic domain can signal midline repulsion to the same extent as Robo1 when combined with the Robo1 ectodomain. We show that SAX-3 is not subject to endosomal sorting by the negative regulator Commissureless (Comm) in Drosophila neurons in vivo, and that peri-membrane and ectodomain sequences are both required for Comm sorting of Drosophila Robo1.     

Recording fox baiting effort across the landscape using geographic information systems: Facilitating more effective management

Predation on native vertebrates and livestock by the European Red Fox (Vulpes vulpes) remains a significant problem in many parts of Australia. Coordinated approaches to fox control are most effective in protecting these assets and involve placement of baits across the landscape by private and public land managers. However, participation in such programmes varies seasonally and the spatial coverage of baiting is often difficult to determine. Here, we describe a geographic information systems‐based system that assists land managers to collect and use spatial information, minimizing gaps in bait coverage and maximizing bait encounters by foxes to increase the effectiveness of pest control. The coordination of data collection and reporting between land managers should facilitate more effective adaptive management by allowing better strategic planning and increasing landholder involvement, which should, in turn, improve the programme's efficacy, provided other critical conditions and resources are met.