Thermobiosensors: features of design and function and prospects for their use

Thermobiosensors are biosensors, which utilize thermal sensors (thermistors, thermocouple) for measuring changes of heat arising during ferment, immune or metabolic reactions. Thermosensors apply to an estimation of a condition of ferment systems, interactions of immune components, kinetics of a biocatalysis, and also for application in biochemical diagnostics, at definition of quality of products, monitoring of an environment and control of biotechnological processes. In the review the data, concerning of the major principle of development and creation mini- and microthermobiosensors, are given on the basis of modern achievement in the field of technology of micromanufacturing. The various types mini-, micro- and multithermobiosensors, and also their application in separate areas of the biochemical and clinical analysis and monitoring of an environment are surveyed. The development and modern achievement in creation of separate variants thermosensors is in detail described on the basis of thermistors and thermocouples. The place thermosensors in general development of biosensor controls is allocated and the attention to prospect of their further improvement and application is given.     

Animal cell cultures in microsporidial research: their general roles and their specific use for fish microsporidia

The use of animal cell cultures as tools for studying the microsporidia of insects and mammals is briefly reviewed, along with an in depth review of the literature on using fish cell cultures to study the microsporidia of fish. Fish cell cultures have been used less often but have had some success. Very short-term primary cultures have been used to show how microsporidia spores can modulate the activities of phagocytes. The most successful microsporidia/fish cell culture system has been relatively long-term primary cultures of salmonid leukocytes for culturing Nucleospora salmonis. Surprisingly, this system can also support the development of Enterocytozoon bienusi, which is of mammalian origin. Some modest success has been achieved in growing Pseudoloma neurophilia on several different fish cell lines. The eel cell line, EP-1, appears to be the only published example of any fish cell line being permanently infected with microsporidia, in this case Heterosporis anguillarum. These cell culture approaches promise to be valuable in understanding and treating microsporidia infections in fish, which are increasingly of economic importance.     

Disruptive coloration provides camouflage independent of background matching

Natural selection shapes the evolution of anti-predator defences, such as camouflage. It is currently contentious whether crypsis and disruptive coloration are alternative mechanisms of camouflage or whether they are interrelated anti-predator defences. Disruptively coloured prey is characterized by highly contrasting patterns to conceal the body shape, whereas cryptic prey minimizes the contrasts to background. Determining bird predation of artificial moths, we found that moths which were dissimilar from the background but sported disruptive patterns on the edge of their wings survived better in heterogeneous habitats than did moths with the same patterns inside of the wings and better than cryptic moths. Despite lower contrasts to background, crypsis did not provide fitness benefits over disruptive coloration on the body outline. We conclude that disruptive coloration on the edge camouflages its bearer independent of background matching. We suggest that this result is explainable because disruptive coloration is effective by exploiting predators' cognitive mechanisms of prey recognition and not their sensory mechanisms of signal detection. Relative to disruptive patterns on the body outline, disruptive markings on the body interior are less effective. Camouflage owing to disruptive coloration on the body interior is background-specific and is as effective as crypsis in heterogeneous habitats. Hence, we hypothesize that two proximate mechanisms explain the diversity of visual anti-predator defences. First, disruptive coloration on the body outline provides camouflage independent of the background. Second, background matching and disruptive coloration on the body interior provide camouflage, but their protection is background-specific.     

Microhabitat use and hindwing phenotype in Hipparchia semele (Lepidoptera, Satyrinae): thermoregulation and background matching

Abstract. 1. Microhabitat use and background matching by adult Hipparchia semele (L.) in a coastal sand dune site in south Wales were studied during the flight period of 1986.
2. Microhabitat use by both sexes is related to temperature and activity. When settling substrates are cool, typically during the early morning, most individuals are located on the warmest basking substrate, lichen patches. After a period of warming, males perch to locate mates on exposed areas of sand where there is a large field of view and where they are visually apparent. Egg-laying females tend to settle on lichen patches throughout the day.
3. Overall, females are maximally visually cryptic when settled on lichen patches with unexposed forewings. Male crypsis on lichen is less effective, but greater on most other substrates, than that of females. It is argued that individuals match lichen patches because this substrate is used when they are most liable to detection and capture by vertebrate predators. Male crypsis is probably a compromise between maximizing protection on lichen patches and requirements for resemblance to other substrates.
4. The underside hindwing phenotype does not match sand. Background matching to this substrate is probably not important because it tends to be used when individuals are active and can effectively use secondary defence mechanisms.
5. It is suggested that the apical eyespot and orange patch on the forewing underside have a dual role in secondary defence, acting as a startling and deflective device, being exposed by disturbed individuals and by those engaged in activities which may disrupt the visual crypsis afforded by the hindwing underside.     

Isolates and their potential use in complex gene mapping efforts

Linkage disequilibrium (LD), detectable with microsatellites in disease alleles over wide genetic intervals in population isolates, has facilitated mapping and positional cloning of numerous disease genes. We, among others, have shown that the LD intervals reach up to 1 Mb in general alleles of young subisolates, and that this feature most probably offers an avenue for the initial locus positioning for complex traits. Development of efficient SNP genotyping and characterization of haploblock structure of the human genome have introduced new prospects to LD-based fine mapping and haplotype-association studies. Encouraging associations have been reported for several complex diseases. Final breakthroughs in mapping of complex disease loci have emerged on large pedigrees in population isolates. Conversely, ignoring genealogical makeup of the study population seems to disclose false negative and false positive associations, directing resources down the drain.     

Chemical features of flavonols affecting their genotoxicity. Potential implications in their use as therapeutical agents

Flavonls are natural compounds present in edible plants and possess several biological activities that can be useful in drug design. Conversely some of these compounds have been shown to be genotoxic to prokaryotic and eukaryotic cells. In this study we tried to establish the chemical features responsible for the genotoxicity of flavonols and to study the conditions that can modulate their genotoxicity namely pH, the presence of antioxidants and metabolism. We assessed the induction of revertants in Salmonella typhimurium TA98 and the induction of Chromosomal aberrations in V79 cells by eight different flavonols and one catechin in the presence and in the absence of metabolizing systems. We have also studied the generation of hydroxyl radical by these flavonoids using the deoxyribose degradation assay. The results obtained in this study suggest that flavonols having a free hydroxyl group at position 3 of the C ring, a free hydroxyl group at position 7 of the A ring and a B ring with a catechol or pyrogallol structure, or a structure that after metabolic activation is transformed into a catechol or a pyrogallol, are flavonols whose genotoxicity in eukaryotic cells depends on their autooxidation. These flavonols can autooxidize when the pH value is slightly alkaline, such as in the intestine, and therefore can induce genotoxicity in humans. Given the above mentioned considerations it is necessary to clarify the mechanisms and the conditions that mediate the biological effects of flavonols before considering them as therapeutical agents.     

Evaluation of DNA and nuclear protein features for their use in describing normal and malignant endometrium

Different feature sets (geometric, densitometric, and textural) derived from DNA and nuclear protein staining were evaluated for their use in describing atrophic, secretory, and proliferative endometrium, and well-differentiated stage I and moderately differentiated stage I adenocarcinomas of the endometrium. It was found that the pattern of significant differences among these groups varied between feature sets, while remaining consistent within a set of features. The DNA density and run-length features were not very effective in describing group mean differences, whereas the co-occurrence features revealed significant differences among most groups. The protein run-length features were the only ones that consistently showed a difference between proliferative endometrium and well-differentiated adenocarcinomas. Analyses repeated on only cells in the G0/G1 DNA region improved the differentiation between moderately differentiated adenocarcinomas and the other groups. It was concluded that the use of DNA and nuclear protein texture features are effective in describing group differences that cannot be described by DNA content only.     

Synthesis of isotope labeled oligonucleotides and their use in an NMR study of a protein-DNA complex.

The synthesis of an oligonucleotide labeled with 13C at the thymine methyl and 15N at the exocyclic amino groups of the cytosines is described. 13CH3I and 15NH4OH were used as sources of the labels. The labeled oligonucleotide was characterized by several NMR techniques. The duplex possesses a labeled functional group in the major groove at every base pair which makes it a very suitable probe for the study of sequence-specific protein-DNA interaction. The labeled thymine methyl group facilitates the detection of hydrophobic contacts with aliphatic side-chains of proteins. This is demonstrated in an NMR study of a complex between the glucocorticoid receptor DNA-binding domain and the labeled oligomer, which revealed a hydrophobic contact between a thymine methyl group and the methyl groups of a valine residue. There are indications for small differences between the solution structure the X-ray structure of the complex.     

The assessment of airborne microorganisms in large-scale composting facilities and their immediate surroundings

The number of airborne microorganisms in the area of large-scale composting facilities with different composting techniques (A: open facility using the intensive decomposition process [4000 t/year]. B: closed facility with compost containers [7000 t/year], C: closed facility with table-pile compositing and automatic turning equipment [22 000 t/year]) was investigated using impactor sampling systems (Andersen samplers). All counts carried out inside the closed facilities, especially during the turning process, showed values of >5.0 × 10⁵ CFU/m³ for viable bacteria and moulds with a proportion ofAspergillus fumigatus of up to 64%. Depending on the type of facility, different median values were determined inside the plant area. Counts were highest in the immediate area around the biofilter outside of Facility C (1.7 × 10⁴ CFU/m³ for bacteria and 9.5 × 10³ CFU/m³ for moulds). In view of the high load of ambient airborne microorganisms inside the composting facilities, adequate occupational health measures are urgently required. Counts determining the hazard to neighbourhood residents at distances of between 150 and 2000 m showed, depending on the facility, annual median values of 170–330 CFU/m³ for bacteria, 75–340 CFU/m³ for moulds, and 15–52 CFU/m³ forA. fumigatus. Higher individual counts — up to 3 × 10³ CFU/m³ for moulds and up to 350 CFU/m³ forA. fumigatus — were found as a result of specific climatic influences, (e.g. winds) and activities as well poor operation. Given the high proportion ofA. fumigatus in the exhaust air, this mould can serve as an indicator for the evaluation of the health risk. However, the maximum values found in the present study, may also be caused by other events in rural areas, (e.g. agricultural activities). With regard to neighbourhood residents, odour complaints are more important than pollution by microorganisms.     

Song types and their structural features are associated with specific contexts in the banded wren

We studied the use of song types and their acoustic features in different social contexts in the banded wren (Thryothorus pleurostictus), a resident tropical songbird in which males possess about 20 distinctive song types varying in duration, bandwidth, note composition, and trill structure. We recorded six focal males intensively for four days each while we observed context information such as during versus after dawn chorus, presence of the female, counter-versus solo-singing, location at the edge versus centre of the territory, and proximity to the nest. All males used at least some song types differentially during each of these pairs of alternative contexts. Males also preferentially used the song types they shared with a given neighbour when interacting with that bird. Songs delivered during dawn chorus were significantly longer, wider in bandwidth, often compound (double songs), and more likely to contain a rattle or buzz and an up-sweeping trill, compared to songs delivered after dawn chorus. Similar features were also more commonly observed when birds were engaged in intense male-male interactions and boundary disputes after dawn chorus, especially when countersinging at the edge of the territory. The presence of the female caused the male to deliver song types with narrower whole-song and trill bandwidth and fewer rattles and buzzes, and song-type diversity and fraction of compound songs were higher when the female was present. Thus, in addition to using type matching and variations in song-type switching and diversity to signal different levels of aggressive intention, male banded wrens also select song types based on their acoustic structure in different social contexts.     

Stable [57Co]-bleomycin complex with a very high specific radioactivity for use at very low concentrations.

A method for the preparation of 57Co-labelled bleomycin (BLM) possessing very high specific radioactivity and suitable for use at the nanomolar concentration range is described, and validated using a biological assay. Chelation of BLM with Co(II) results in a very stable complex. However, association does not occur below the micromolar concentration range. A nanomolar [57Co]BLM solution with maximal specific radioactivity can be easily prepared, without handling unreasonable amounts of radioactivity, provided that: equimolar solutions of BLM and [57Co]Cl2 are first mixed at the micromolar concentration range and that the mixture is then diluted a thousand times to reach the nanomolar concentration range.     

The crystal structure of Cys-tRNACys-EF-Tu-GDPNP reveals general and specific features in the ternary complex and in tRNA.

BACKGROUND:. The translation elongation factor EF-Tu in its GTP-bound state forms a ternary complex with any aminoacylated tRNA (aa-tRNA), except initiator tRNA and selenocysteinyl-tRNA. This complex delivers aa-tRNA to the ribosomal A site during the elongation cycle of translation. The crystal structure of the yeast Phe-tRNAPhe ternary complex with Thermus aquaticus EF-Tu-GDPNP (Phe-TC) has previously been determined as one representative of this general yet highly discriminating complex formation. RESULTS: The ternary complex of Escherichia coli Cys-tRNACys and T. aquaticus EF-Tu-GDPNP (Cys-TC) has been solved and refined at 2.6 degrees resolution. Conserved and variable features of the aa-tRNA recognition and binding by EF-Tu-GTP have been revealed by comparison with the Phe-TC structure. New tertiary interactions are observed in the tRNACys structure. A 'kissing complex' is observed in the very close crystal packing arrangement. CONCLUSIONS: The recognition of Cys-tRNACys by EF-Tu-GDPNP is restricted to the aa-tRNA motif previously identified in Phe-TC and consists of the aminoacylated 3' end, the phosphorylated 5' end and one side of the acceptor stem and T stem. The aminoacyl bond is recognized somewhat differently, yet by the same primary motif in EF-Tu, which suggests that EF-Tu adapts to subtle variations in this moiety among all aa-tRNAs. New tertiary interactions revealed by the Cys-tRNACys structure, such as a protonated C16:C59 pyrimidine pair, a G15:G48 'Levitt pair' and an s4U8:A14:A46 base triple add to the generic understanding of tRNA structure from sequence. The structure of the 'kissing complex' shows a quasicontinuous helix with a distinct shape determined by the number of base pairs.     

Mineral impact of lakes as background for their biogenic dynamics

Favourable circumstances for limnological observations were offered by human activities at shores of some Holstein lakes. The lakes were enriched by clay, silt, sandy particles and hydroxides of iron and manganese over many years in consequence of an embankment built at Schöhsee and influx of washing water from a gravel pit, respectively. During windy weather the lakes were extremely turbid, but very clear during calm weather. The 1% penetration level of green light was found at 11 and 14 m. Nutrients as well as small phytoplankton cells and bacteria were adsorbed on to the detritus. Therefore, the sedimentation rate increased and consequently that of phytoplankton production decreased. The sediments became rich in minerals comparable with alpine lakes. The epilimnic decomposition rate was much lower than in normal Holstein lakes in the neighbourhood, as in Groer Plöner See and the highest value 15 of the Short-Circuit-Metabolism (SCM) was measured in Pluss-See, i.e., there occurred 15 times re-use of carbon (and other) molecules in the euphotic region before settling down. The conclusion drawn is that the mineral impact of alpine lakes occurring annually under natural conditions, at least in spring, is one of the most important factors of minor bioactivity compared with eutrophic lowland lakes.