A three channel radio-controlled brain-stimulator for unrestrained animals

We have developed a project for biological radio-controlled electrostimulation, consisting in a three channel telestimulator, which has been used in our laboratories on animals unrestrained and complete freedom of movements. The system has been tested and used in many experiments which excellent results.     

Exploring bird aerodynamics using radio-controlled models

A series of radio-controlled glider models was constructed by duplicating the aerodynamic shape of soaring birds (raven, turkey vulture, seagull and pelican). Controlled tests were conducted to determine the level of longitudinal and lateral-directional static stability, and to identify the characteristics that allowed flight without a vertical tail. The use of tail-tilt for controlling small bank-angle changes, as observed in soaring birds, was verified. Subsequent tests, using wing-tip ailerons, inferred that birds use a three-dimensional flow pattern around the wing tip (wing tip vortices) to control adverse yaw and to create a small amount of forward thrust in gliding flight.     

A gene trap approach in Xenopus.

The frog transgenesis technique ultimately promises to make mutagenesis possible through random insertion of plasmid DNA into the genome. This study was undertaken to evaluate whether a gene trap approach combined with transgenesis would be appropriate for performing insertional mutagenesis in Xenopus embryos. Firstly, we confirmed that the transgenic technique results in stable integration into the genome and that transmission through the germline occurs in the expected Mendelian fashion. Secondly, we developed several gene trap vectors, using the green fluorescent protein (GFP) as a marker. Using these vectors, we trapped several genes in Xenopus laevis that are expressed in a spatially restricted manner, including expression in the epiphysis, the olfactory bulb and placodes, the eyes, ear, brain, muscles, tail and intestine. Finally, we cloned one of the trapped genes using 5' rapid amplification of cDNA ends polymerase chain reaction (RACE PCR). These results suggest that the transgenic technique combined with a gene trap approach might provide a powerful method for generating mutations in endogenous genes in Xenopus.     

A size selective underwater light trap

A size selective underwater light trap is described. Trap records indicate that the trap is effective in taking a wide variety of organisms within size limits that can be set by the experimenter.     

Genetically engineered live attenuated influenza A virus vaccine candidates.

We have generated new influenza A virus live attenuated vaccine candidates by site-directed mutagenesis and reverse genetics. By mutating specific amino acids in the PB2 polymerase subunit, two temperature-sensitive (ts) attenuated viruses were obtained. Both candidates have 38 degrees C shutoff temperatures in MDCK cells, are attenuated in the respiratory tracts of mice and ferrets, and have very low reactogenicity in ferrets. Infection of mice or ferrets with either mutant conferred significant protection from challenge with the homologous wild-type virus. Three tests for genetic stability were used to assess the propensity for reversion to virulence: 14 days of replication in nude mice, growth at 37 degrees C in tissue culture, and serial passage in ferrets. One candidate, which contains mutations intended to reduce the ability of PB2 to bind to cap structures, was stable in all three assays, whereas the second candidate, which contains mutations found only in other ts strains of influenza virus, lost its ts phenotype in the last two assays. This approach has therefore enabled the creation of live attenuated influenza A virus vaccine candidates suitable for human testing.     

A signal sequence trap based on a constitutively active cytokine receptor.

Targeting of secreted and cell-surface proteins to the cell membrane is mediated by a short hydrophobic stretch of amino acids, termed the signal sequence. We have developed a method that detects signal sequences in cDNA fragments based on their ability to redirect a constitutively active mutant of a cytokine receptor to the cell surface, thereby permitting interleukin-3 (IL-3)-independent growth of Ba/F3 cells. Retrovirus-mediated expression of the fusions in IL-3-dependent cells was followed by selection of clones for growth in the absence of IL-3. Infection of cells with 5x10(6) viral particles in a pilot experiment led to the isolation of 150 known and 48 novel cDNA clones, and all the known cDNA clones were found to encode secreted and cell-surface proteins. In addition, we isolated type II membrane proteins, which have not been detected by existing signal sequence trap strategies.     

A new signal sequence trap using alkaline phosphatase as a reporter.

Secreted and transmembrane proteins are critical to the cell-cell interactions governing normal development and carcinogenesis. To facilitate the identification of such molecules, we have developed a novel signal sequence trap that uses human placental alkaline phosphatase as a reporter. Libraries from mouse prostate and human prostatic carcinoma were constructed to test the PST (peptide signal trap) system, resulting in the identification of several secreted and transmembrane proteins.     

A carbon dioxide trap for prolonged sampling ofIxodes ricinus L. populations

A carbon dioxide trap designed to captureIxodes ricinus over periods of up to 7 days is described. The trap compared favourably with blanket dragging and flagging in areas of high tick density, particularly on rough ground and for adult ticks; it was also surprisingly efficient for larvae. Ticks appeared to be captured throughout the 7-day trapping period. The maximum attraction distance recorded for adult female ticks was 3.5 m and for nymphs 1.0 m. Trapping rates were influenced by air temperature.     

A trap for capturing planktonic chironomid larvae

This paper describes the construction and field operation of a trap designed to investigate the movements of larval Chironomidae in the open water of lakes. Although the trap was designed to be multidirectional, with six trapping bottles, only two directional components, vertical and horizontal, could be distinguished. This led to examination of three bottles only during later field studies, namely bottle 1 facing into the current, bottle 5 facing the substrate and bottle 6 facing the surface of the lake. Some results are presented and trap operation and sampling efficiency discussed. A major advantage of the trap is its use as a long term accumulator of planktonic organisms as opposed to traditional spot sampling techniques which tend to miss the less numerous plankters.     

Long live the Queen's subjects.

In 1952, the Queen congratulated 255 people on their hundredth birthdays and 1135 couples on their sixtieth wedding anniversaries. By 1996, these numbers had risen to 5218 and 11,688, respectively. Semilogarithmic plots, normalized to constant numbers of births and marriages, show steady exponential rises in the number of centenarians with a doubling time of 11 years, and of diamond weddings with a doubling time of 19 years. An alternative plot of the numbers of those reaching a hundred between 1910 and 1990, based on registers of births and deaths and normalized to constant births, shows an annual rise of only 1% from 1910 to 1946, followed by a steady exponential rise with a doubling time of 12 years, closely matching that of 11 years derived from the Queen''s figures. The exponential rise in the number of those born from 1846 onwards living to a hundred precedes by many years the general rise in the expectation of life at birth and the general drop in mortality from infectious diseases, but it coincides with the beginning of a steady rise in real wages. Another important factor may be improved medical treatment at old age from 1946 onwards.     

Genetic live vaccines mimic the antigenicity but not pathogenicity of live viruses.

The development of an effective HIV vaccine is both a pressing and a formidable problem. The most encouraging results to date have been achieved using live-attenuated immunodeficiency viruses. However, the frequency of pathogenic breakthroughs has been a deterrent to their development. We suggest that expression libraries generated from viral DNA can produce the immunologic advantages of live vaccines without risk of reversion to pathogenic viruses. The plasmid libraries could be deconvoluted into useful components or administered as complex mixtures. To explore this approach, we designed and tested several of these genetic live vaccines (GLVs) for HIV. We constructed libraries by cloning overlapping fragments of the proviral genome into mammalian expression plasmids, then used them to immunize mice. We found that inserting library fragments into a vector downstream of a secretory gene sequence led to augmented antibody responses, and insertion downstream of a ubiquitin sequence enhanced cytotoxic lymphocyte responses. Also, fragmentation of gag into subgenes broadened T-cell epitope recognition. We have fragmented the genome by sequence-directed and random methods to create libraries with different features. We propose that the characteristics of GLVs support their further investigation as an approach to protection against HIV and other viral pathogens.