Paired assessment of volatile anesthetic concentrations with synaptic actions recorded in vitro

The volatile anesthetic isoflurane poses a number of experimental challenges in the laboratory. Due to its rapid evaporation, the open conditions of most in vitro electrophysiological recording systems make the determination of actual isoflurane concentrations a challenge. Since the absolute anesthetic concentration in solution is directly related to efficacy, concentration measurements are important to allow comparisons between laboratory and clinical studies. In this study we quantify the sources of isoflurane loss during experimentation and describe a method for the measurement of isoflurane concentrations using gas chromatography and mass spectrometry simultaneous to in vitro electrophysiological measurements. Serial samples of perfused bath solution allowed correlation of isoflurane concentrations with ongoing biological effects. Saturated physiological solutions contained 13.4 +/- 0.2 mM isoflurane and were diluted to desired "nominal" concentrations for experiments. The perfusion system established stable isoflurane concentrations within the bath by 2 minutes. However, bath isoflurane concentrations varied substantially and unpredictably between experiments. The magnitudes of such discrepancies in isoflurane concentrations spanned clinically important levels. Our studies suggest that, despite countermeasures, solution handling significantly impacted the isoflurane content in the tissue bath. The magnitude of these discrepancies appears to necessitate systematic direct measurement of bath isoflurane concentrations during most in vitro conditions.     

Plant invasions in treeless vegetation of the Australian Alps

A total of 128 invasive plant species have been recorded in treeless vegetation in the Australian Alps. Most of these are forbs and most are uncommon. Cover of invasive species is generally minimal unless there has been gross disturbance to natural vegetation and soils. Although there is a significantly positive correlation between invasive and native species diversity, suggesting that conditions that allow numerous native species to co-exist also permit more plant invasions, altitude is the most important determinant of invasive species diversity. Only 22 of the 128 species have been recorded above 1800 m. Some plant communities (e.g. those with high pH or relatively nutrient-rich soils), however, seem to be vulnerable to invasions regardless of altitude. Most invasive species appear to have been introduced unintentionally (e.g. as seed attached to vehicles, animals and humans) but a few were introduced to assist with revegetation of disturbed soils and for amenity plantings in ski resorts, and have subsequently established in native vegetation. Treeless communities in the Australian Alps are likely to face increasing pressure from invasive species as a result of global warming and continued introduction of non-native plants to ski resort gardens. Whilst it may be difficult to prevent invasive species of low elevations migrating to higher elevations as temperatures rise, the risk of invasion from garden plants could be minimised through regulation. Non-native plants in ski resort gardens pose a far greater risk than most invasive species currently present in the Alps because they have been chosen for their capacity to survive at high altitudes.     

Design, synthesis, and antiviral evaluation of some 3'-carboxymethyl-3'-deoxyadenosine derivatives

3'-Carboxymethyl-3'-deoxyadenosine derivatives were prepared from 2'-O-TBDMS-3'-[(ethoxycarbonyl)methyl]-3'-deoxyadenosine (1) via simple and efficient procedures. Conversion of 1 to its 5'-azido-5'-deoxy derivative 5 was accomplished via a novel one-pot method employing 5'-activation (TosCl) followed by efficient nucleophilic displacement with tetramethylguanidinium azide. Compound 5 was converted to 5'-[(N-methylcarbamoyl)amino] derivative 8 via one-pot reduction/acylation employing H(2)/Pd-C followed by treatment with p-nitrophenyl N-methylcarbamate. N(6)-phenylcarbamoyl groups were introduced by treatment with phenylisocyanate, and an efficient new method for lactonization of 2'-O-TBDMS-3'-[(ethoxycarbonyl)methyl]-3'-deoxyadenosines to give corresponding 2',3'-lactones was also developed. Target compounds were evaluated for anti-HIV and anti-HIV integrase activities, but were not active at the concentrations tested.     

Adenovirus 12 uncoiler regions of human chromosome 1 in relation to the 5S rRNA genes

Two uncoiler regions, induced by adenovirus 12, have been identified on human chromosome 1 at 1q42 and 1p36. In situ hybridization with [¹²⁵I-5S]-rRNA places the 5S genes more accurately at 1q42–43 immediately distal to the uncoiled site, 1q42.     

In vitro traits of adenovirus-transformed cell lines and their relevance to tumorigenicity in nude mice

Six independently isolated adenovirus 2-transformed rat cell lines and one adenovirus 5-transformed human cell line have been examined in vitro for serum growth requirements, saturation density, anchorage-independent growth, proteolytic enzyme activity and the presence of LETS glycoprotein and T antigen. This series of adenovirus-transformed cell lines exhibits an oncogenic spectrum ranging from being tumorigenic in immunocompetent rats through to nontumorigenic in adult nude mice. The relevance of the in vitro findings to growth potential in vivo is discussed.     

Enhanced operation of femtosecond lasers and applications in cell transfection

In this work we present a review and discussion on the enhancement of femtosecond (fs) lasers for use within biophotonics with a particular focus on their use in optical transfection techniques. We describe the broad range of source options now available for the generation of femtosecond pulses before briefly reviewing the application of fs laser in optical transfection studies. We show that major performance enhancements may be obtained by optimising the spatial and temporal performance of the laser source before considering possible future directions in this field. In relation to optical transfection we describe how such laser sources initiate a multiphoton process to permeate the cell membrane in a transient fashion. We look at aspects of this technique including the ability to combine transfection with optical trapping. For future implementation of such transfection we explore the role of new sources and “nondiffracting” light fields. (© 2008 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)