Occupational (In)visibility: The emerging role of the Remote Education Tutor as an educational conduit.

Remote Education Tutors (RETs) are central to the delivery of distance schooling in Australia and are accountable for the face-to face supervision and educational support of students. They act as the government mandated adult supervisors of Australian primary and secondary school students enrolled in distance education, including geographically isolated learners. This paper draws on statistical data from a national survey (N = 575) that was designed to map the perceptions of Australian RETs. These data confirmed that RETs act as a conduit between the distance schooling teacher and student, and that their role requires complex capabilities to be performed within a structured framework. Time restrictions with competing demands present a constant challenge to the RETs' work satisfaction. Constraining this occupation is the reality that there is no formal qualification available for RETs. Without specific credentialling, it appears that the RETs' (in)visible role risks being overlooked as a substantive educational occupation.

Biomarkers of heavy metal contamination in the red fingered marsh crab, Parasesarma erythodactyla.

Variation in glutathione antioxidant biochemistry in response to metal contamination and accumulation under field conditions was examined in the brachyurid grapsid, Parasesarma erythodactyla. Significant relationships suggesting accumulation were found between sediment metals and metals in crab tissue for Pb, Cu, Cr, Zn, and Se in males and Cd, Pb, Cr Zn, As, and Se in females. Higher pH and lower organic content were associated with greater uptake of selected metals in males and females. Higher salinity was related to increased metal uptake for Cu and Zn in males and lower salinities to increased Se uptake for males and females. When examining metals, which were elevated in crabs, patterns of site discrimination were similar to sediment metal site discrimination for both males and females. In terms of biochemical responses, glutathione levels remained constant while glutathione peroxidase activity was elevated in individuals where metals were elevated. Only females with the highest levels of accumulated metals exhibited increases in lipid peroxidation products. Glutathione peroxidase activity may be a sensitive biomarker of metal exposure and biological effect and lipid peroxides as a secondary marker when accumulated metals are high.

The development of a microimmunoadsorbent assay and a microdialysis method for the evaluation of immunoadsorbent efficiency and antibody quantitation.

A microimmunoadsorbent assay which provides for the quantitative elution and recovery of active antibody from immunoaffinity columns has been developed. We have also designed a microdialysis chamber which permits the dialysis of sample volumes from 50 to 500 microliters and allows quantitative recovery of macromolecular compounds. These techniques permit the rapid evaluation of a large number of eluants and immunoadsorbents in a short period of time. The microimmunoadsorbent assay coupled with the microdialysis method has been used to evaluate the efficacy of different anti-isopentenyl adenosine immunoadsorbents under various elution conditions. In the present study the best eluants for immunoaffinity purification of anti-N6-(delta 2-isopentenyl)adenosine antibody contained 15% pyridine in a sodium phosphate buffer or 4 M imidazole-HCl at pH 10.0.

Characterization of monoclonal antibodies specific for isopentenyl adenosine derivatives occurring in transfer RNA.

A family of isopentenyl adenosine derivatives are naturally occurring components of transfer RNA and are involved in several different functional roles in the cell. To facilitate the study of the biochemistry of these modified nucleosides we have raised monoclonal antibodies to N6-(delta 2-isopentenyl)adenosine and N6-(4-hydroxy-3-methyl-but-2-enyl)adenosine. The antibodies show considerable specificity and three characteristic types are distinguishable. The first type have the hydroxylated derivative as the preferred antigen, the second type have isopentenyl adenosine as the preferred antigen and a third type show a specificity for all isopentenyl-containing derivatives.

Investigation of immunoadsorbent efficiency and capacity for the isolation of tRNAs containing N6-(delta 2-isopentenyl)adenosine derivatives.

Antibodies directed to modified nucleosides recognize the nucleoside (antigen) when it is present in an intact tRNA molecule. The general application of anti-nucleoside immunoadsorbent chromatography, however, has been greatly impeded by the apparent inefficiency and low capacity of conventional immunoadsorbents for transfer RNA. Antibodies specific for isopentenyladenosine (i6A) were employed to investigate the efficacy of various immunoadsorbents with respect to immobilization of antibody protein and with respect to their ability to bind i6A-containing tRNAs. Biologically active anti-i6A was recovered in high yield (80-88%) by affinity chromatography on i6A-adipate-Sepharose 4B or i6A-butane diglycidyl ether-Sepharose 4B using either 15% pyridine in phosphate-buffered saline or 0.2 M acetic acid as eluents. The binding capacity of various anti-i6A antibody immunoadsorbents was evaluated. While both anti-i6A antibody-protein A-agarose-iminothiolane (ITL) and anti-i6A antibody-protein A-agarose-dimethyl suberimidate showed a high capacity for i6A-tRNA, the latter column is much less efficient with respect to antibody immobilization. Under optimal conditions, the ITL immunoadsorbent binds 5-6 nmol of i6A/mg of antibody protein. With respect to bulk tRNA, 1 mg of antibody protein (ITL immunoadsorbent) binds all of the i6A-tRNA in a 1-mg sample.

The transfer RNA of certain Enterobacteriacae contain 2-methylthiozeatin riboside (ms2io6A) an isopentenyl adenosine derivative.

Isopentenyl adenosine derivatives are always located adjacent to the 3' end of the anticodon in transfer RNA and have been implicated in certain biological functions. In the enteric bacterium, E. coli, the derivative is ms2i6A whereas in some plant associated bacteria the derivative is the hydroxylated form, ms2io6A. Anti-i6A immunoadsorbent chromatography has been employed to detect isopentenyl adenosine compounds. In the present study we show that the transfer RNA of three species of enteric bacteria, S. typhimurium, K. pneumoniae, and S. marcescens contains both ms2io6A and ms2i6A. Under the growth conditions utilized the ms2io6A is predominant. The presence of ms2io6A in Enterobacteriacae is particularly noteworthy since in previous work it has been found only in plant-associated species of bacteria.

Iron-related modification of bacterial transfer RNA.

Transfer RNAs isolated from E. coli grown in media where ferric iron is not freely available show well characterized chromatographic changes due to the absence of the methylthio moiety of ms2i6A. The altered tRNA molecules include tRNA trp tRNA tyr, tRNA phe and two minor tRNA ser species. It has been suggested that methylthiolation of tRNA affects its function in regulation. We now show iron-related changes in tRNA trp from S. typhimurium, Ps. aeruginosa and K. pneumoniae. tRNA trp from S. typhimurium contains ms2i6A and it seems probable that the availability of iron affects the synthesis of ms2i6A-tRNA trp from i6A-tRNA trp in this organism. An iron-related methylthiolating system may also be operative in K. pneumoniae. S. marcescens tRNA trp, however was not affected by the availability of iron. Neither ms2i6A nor i6A was found in S. marcescens tRNA, although an, as yet unidentified, hydrophobic nucleoside was present.

A modified quantitative precipitin test which is rapid, sensitive and reproducible.

A modification of the standard quantitative precipitin test is described. The new procedure, which utilizes polyethylene glycol (PEG) at both the 37 degrees C and 4 degrees C incubation stages, provides a convenient quantitative precipitin test assay which is more reliable and much faster than the standard assay. Moreover, the modified or PEG assay gives a quantitative measure of antibody concentration even when antisera of low titre are tested. The sensitivity of the capillary-tube test is also enhanced when the supernatant solutions contain PEG.

N-6-(delta-2-isopentenyl) adenosine: hydrolysis by a mucleosidase isolated from Lactobacillus acidophilus cells.

A nucleosidase activity has been isolated from Lactobacillus acidophilus which rapidly hydrolyses N-6 (delta-2-isopentenyl) adenosine to its corresponding base, N-6(delta-2-isopentenyl) adenine. The activity can be distinguished from the spleen exzyme (EC. 2.4.2.1), a purine nucleoside transferase, on the basis of its substrate specificity, electrophoretic behavior, and nondependence on phosphate. The bacterial enzyme hydrolyzes both inosine and isopentenyl adenosine, giving Km values of 63.3muM and 177 muM respectively. The presence of this enzyme in bacteria counts for the rapid conversion of the parent nucleoside to isopentenyl adenine, which has been observed in these cells. The enzyme thus assumes importance as one of the catabolic activities available to the cell for metabolizing the cytokinin, N-6-(delta-2-isopentenyl) adenosine.

Cytokinins in the soluble RNA of plant tissues.

The cytokinin, N(6)-(Delta(2)-isopentenyl) adenosine occurs in the soluble RNA of yeast and mammalian tissue and has now been detected in plant soluble RNA. A hydroxylated derivative of this cytokinin 6-(cis-4-hydroxy-3-methylbut-2-enylamino)-9-,beta-D-ribofuranosylpurine has also been identified as a constituent of plant soluble RNA.