Aminoacylation of rat liver transfer RNA with homologous and heterologous enzyme systems during aging

Total tRNA extracted from livers of young (7 +/- 1 weeks), adult (40 +/- 1 weeks) and old (80 +/- 1 weeks) rats showed quantitative variation with age, being maximal in adults. Young and old animals yielded almost the same level of tRNAs. Quantitative changes in tRNAs were also observed from the study of amino acid acceptor activity using homologous enzyme i.e., aminoacyl-tRNA synthetase preparations from rat liver of the same age group. Quantitative variation followed the trend of qualitative variation. When tRNA was amino-acylated with a heterologous enzyme system, i.e., synthetase preparation from rat liver of another age group, age-related variation in aminoacyl-tRNA did not follow a pattern similar to that in the case of the homologous enzyme system. Young and adult synthetase enzymes showed maximum affinity for their homologous tRNAs but synthetases from old rat liver did not show any specific affinity for "old" tRNAs. This shows that apart from tRNAs, enzyme activity also changes with age.     

Specific-primer-directed DNA sequencing using automated fluorescence detection.

Automated fluorescence-based DNA sequence analysis offers the possibility to undertake very large scale sequencing projects. Directed strategies, such as the specific-primer-directed sequencing approach ('gene walking'), should prove useful in such projects. Described herein is a study involving the use of this approach in conjunction with automated fluorescence detection on a commercial instrument (ABI 370A DNA sequencer). This includes procedures for the rapid chemical synthesis and purification of labeled primers, the design of primer sequences that are compatible with the commercial analysis software, and automated DNA sequence analysis using such primers. A set of four fluorophore-labeled primers can be reliably synthesized in a twenty-four hour period, and greater than 300 nucleotides of analyzed new sequence obtained using this set in an additional twenty-four hours. Scale-up of these procedures to take advantage of the full capabilities of the sequencer is, at present, too slow and costly to be suitable for routine sequencing, and therefore the use of specific-primers is best suited to the closure of gaps in extended sequence produced using random cloning and sequencing strategies.     

Independent Mobility Achieved through a Wireless Brain-Machine Interface

Individuals with tetraplegia lack independent mobility, making them highly dependent on others to move from one place to another. Here, we describe how two macaques were able to use a wireless integrated system to control a robotic platform, over which they were sitting, to achieve independent mobility using the neuronal activity in their motor cortices. The activity of populations of single neurons was recorded using multiple electrode arrays implanted in the arm region of primary motor cortex, and decoded to achieve brain control of the platform. We found that free-running brain control of the platform (which was not equipped with any machine intelligence) was fast and accurate, resembling the performance achieved using joystick control. The decoding algorithms can be trained in the absence of joystick movements, as would be required for use by tetraplegic individuals, demonstrating that the non-human primate model is a good pre-clinical model for developing such a cortically-controlled movement prosthetic. Interestingly, we found that the response properties of some neurons differed greatly depending on the mode of control (joystick or brain control), suggesting different roles for these neurons in encoding movement intention and movement execution. These results demonstrate that independent mobility can be achieved without first training on prescribed motor movements, opening the door for the implementation of this technology in persons with tetraplegia.     

A comparison of tRNA populations of rat liver and skeletal muscle during aging

Total tRNA extracted from liver as well as from skeletal muscle of young, adult and old female albino rats showed quantitative variation with age. The amount of liver total tRNA was maximum in adult rats when compared to that in young and old ones, whose levels were almost the same. Transfer RNA from skeletal muscle showed a different pattern with age. It was maximum in young rats and showed a gradual decline with age. Transfer RNAs were aminoacylated using homologous synthetase preparations to study their qualitative variation during aging, which followed the trend of quantitative variation in both the tissues. Arginyl and glutamyl-tRNAs were fractionated from both the tissues at the three ages. Isoacceptor profile of glutamyl-tRNAs showed neither tissue specificity nor age-related change, whereas a definite change was found in the case of arginyl-tRNA isoacceptors in the two tissues during aging.     

Bionectria ochroleuca NOTL33—an endophytic fungus from Nothapodytes foetida producing antimicrobial and free radical scavenging metabolites

Endophytic fungi are reported to produce diverse classes of secondary metabolites. This study investigated the antimicrobial and free radical scavenging activity of a foliar endophytic fungus from Nothapodytes foetida, a medium sized tree known to produce the antineoplastic compound camptothecin. The fungal isolate was identified as Bionectria ochroleuca based on the ITS rDNA analysis. The differences among endophytic, pathogenic and free living Bionectria ochroleuca were established by RNA secondary structure analysis. The metabolites showed a broad spectrum of antibacterial, antifungal and anti-dermatophytic activity. Minimum inhibitory concentration values of ethyl acetate extracts were in the range of 78–625 μg/mL against all test organisms, except for Pseudomonas aeruginosa (5 mg/mL). Antimicrobial components in the ethyl acetate extract were identified by GC-MS analysis. The isolate was also produced volatile antifungal compounds. A dose-dependent free radical quenching was observed in the ethyl acetate extract. This is the first report on Bionectria sp. as an endophyte of N. foetida. The results indicate that the B. ochroleuca NOTL33 isolate is a potential source of antimicrobial agents and could be used as an effective biofumigant.     

Isolation of the 66-kilodalton polypeptide from promastigotes of Leishmania donovani as a ligand molecule for binding to macrophages.

A 66-kDa major plasma membrane-associated molecule of promastigotes of Leishmania donovani (UR6) was purified by affinity chromatography. The immunoreactivity of the 66-kDa molecule was lost upon exposure to heat or treatment with trypsin. The metaperiodate oxidation significantly reduced its immunoreactivity. The 66-kDa molecule is, therefore, glycoprotein in nature. With a fluorescent probe, the 66-kDa molecule was found to be located on the tip of flagellum and on the kinetoplast. The exposure of promastigotes of L. donovani to monospecific anti-66-kDa antibodies significantly reduced the percentage of macrophages with attached promastigotes in the cultured cell line (J774G8). The data suggested that promastigotes of L. donovani utilize the 66-kDa molecule in recognizing and as ligand for binding to macrophages.     

Acid phosphatase activity of promastigotes of Leishmania donovani: a marker of virulence

Seven cloned lines of promastigotes of Leishmania donovani (UR 6) were isolated by limiting dilution. One clone, UR6-C25, failed to multiply inside the macrophages of line J774G8 and thus was labelled as avirulent. Another, UR6-C24, multiplied inside macrophages, had a virulence index as high as 93 +/- 9.8 and was thus labelled as highly virulent. The other five clones had variable degree of virulence indices ranging from 46.4 +/- 5.8 to 67.6 +/- 3.5. No significant difference in the degrees of attachment of virulent and avirulent populations of promastigotes to macrophages was observed, suggesting no difference in the ligand utilised by these populations for attachment to the macrophages. Acid phosphatase activity of cloned promastigotes correlated with the degree of virulence. These data suggest that acid phosphatase activity could be used as a marker to differentiate avirulent from virulent populations of promastigotes of L. donovani.