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Insects as test systems for assessing the potential role of microgravity in biological development and evolution.

Gravity and radiation are undoubtedly the two major environmental factors altered in space. Gravity is a weak force, which creates a permanent potential field acting on the mass of biological systems and their cellular components, strongly reduced in space flights. Developmental systems, particularly at very early stages, provide the larger cellular compartments known, where the effects of alterations in the size of the gravity vector on living organisms can be more effectively tested. The insects, one of the more highly evolved classes of animals in which early development occurs in a syncytial embryo, are systems particularly well suited to test these effects and the specific developmental mechanisms affected. Furthermore, they share some basic features such as small size, short life cycles, relatively high radio-resistance, etc. and show a diversity of developmental strategies and tempos advantageous in experiments of this type in space. Drosophila melanogaster, the current biological paradigm to study development, with so much genetic and evolutionary background available, is clearly the reference organism for these studies. The current evidence on the effects of the physical parameters altered in space flights on insect development indicate a surprising correlation between effects seen on the fast developing and relatively small Drosophila embryo and the more slowly developing and large Carausius morosus system. In relation to the issue of the importance of developmental and environmental constraints in biological evolution, still the missing link in current evolutionary thinking, insects and space facilities for long-term experiments could provide useful experimental settings where to critically assess how development and evolution may be interconnected. Finally, it has to be pointed out that since there are experimental data indicating a possible synergism between microgravity and space radiation, possible effects of space radiation should be taken into account in the planning and evaluation of experiments designed to test the potential role of microgravity on biological developmental and evolution.

Animals↗

EVA dosimetry in manned spacecraft.

Extra Vehicular Activity (EVA) will become a large part of the astronaut's work on board the International Space Station (ISS). It is already well known that long duration space missions inside a spacecraft lead to radiation doses which are high enough to be a significant health risk to the crew. The doses received during EVA, however, have not been quantified to the same degree. This paper reviews the space radiation environment and the current dose limits to critical organs. Results of preliminary radiation dosimetry experiments on the external surface of the BION series of satellites indicate that EVA doses will vary considerably due to a number of factors such as EVA suit shielding, temporal fluctuations and spacecraft orbit and shielding. It is concluded that measurement of doses to crew members who engage in EVA should be done on board the spacecraft. An experiment is described which will lead the way to implementing this plan on the ISS. It is expected that results of this experiment will help future crew mitigate the risks of ionising radiation in space.

Animals↗

Silicone-fat differentiation in the breast: exploiting the bright-fat phenomenon in fast spin-echo MR imaging.

Selective suppression of silicone or fat with chemical shift-selective (CHESS) pulses is difficult because of the small chemical shift difference between the primary lipid signal and the primary silicone signal at 1.5 T. Differentiation of these chemically distinct species is, however, an important clinical task in assessing implant rupture and silicone migration in breast tissue. A method uniquely suited for silicone-fat differentiation with fast spin-echo (FSE) sequences is reported. It is based on the dependence of fat signal on echo spacing in FSE imaging and results show that it may provide a clinically robust method for silicone-fat differentiation in magnetic resonance imaging of the breast.

Adipose Tissue↗

Wavelet domain nonlinear filtering for evoked potential signal enhancement.

A wavelet domain nonlinear filtering method for improving the signal-to-noise ratio (SNR) of the evoked potentials (EP) is proposed. The method modifies the selective filtering technique proposed for edge detection in images by Xu et al. for the case of signals which require a smooth transition at the edge points. It identifies the significant features of a noisy signal based on the correlation between the scales of its nonorthogonal subband decompositions. The signal transition information from interscale correlation coupled with the change in variance around the identified transition region is used to differentiate between noise and the signal. A nonlinear function such as a Gaussian smoothing function applied around the identified edge in the wavelet domain leads to smoothing in the signal space also. Numerical results obtained by applying the proposed nonlinear filtering method on middle latency responses of auditory evoked potentials show that the method is well suited for signal enhancement applications.

Evoked Potentials, Auditory↗

A place that answers questions: primatological field sites and the making of authentic observations.

The ideals and realities of field research have shaped the development of behavioural primatology over the latter half of the twentieth century. This paper draws on interviews with primatologists as well as a survey of the scientific literature to examine the idealized notion of the field site as a natural place and the physical environment of the field as a research space. It shows that what became standard field practice emerged in the course of wide ranging debate about the techniques, personal qualities and site conditions best suited to the scientific study of the natural behaviour of apes and monkeys. Although the laboratory was a constant presence in this debate, the export of techniques from the laboratory to the field was limited, due to concerns that experimental manipulation would destroy the naturalness of the behaviour. The paper goes on to demonstrate the central significance given by primatologists to the unique social, historical and ecological circumstances of particular field sites, and to sketch some of the complexities that fieldworkers contend with in trying to realize their ideals. Primatologists seek field sites that answer their questions; but once their studies become long term, they also need to find questions that answer to ever changing conditions at those sites.

Animals↗

A review and evaluation of intraurban air pollution exposure models.

The development of models to assess air pollution exposures within cities for assignment to subjects in health studies has been identified as a priority area for future research. This paper reviews models for assessing intraurban exposure under six classes, including: (i) proximity-based assessments, (ii) statistical interpolation, (iii) land use regression models, (iv) line dispersion models, (v) integrated emission-meteorological models, and (vi) hybrid models combining personal or household exposure monitoring with one of the preceding methods. We enrich this review of the modelling procedures and results with applied examples from Hamilton, Canada. In addition, we qualitatively evaluate the models based on key criteria important to health effects assessment research. Hybrid models appear well suited to overcoming the problem of achieving population representative samples while understanding the role of exposure variation at the individual level. Remote sensing and activity-space analysis will complement refinements in pre-existing methods, and with expected advances, the field of exposure assessment may help to reduce scientific uncertainties that now impede policy intervention aimed at protecting public health.

Air Pollutants↗

On the use of orientational restraints and symmetry corrections in alchemical free energy calculations.

Alchemical free energy calculations are becoming a useful tool for calculating absolute binding free energies of small molecule ligands to proteins. Here, we find that the presence of multiple metastable ligand orientations can cause convergence problems when distance restraints alone are used. We demonstrate that the use of orientational restraints can greatly accelerate the convergence of these calculations. However, even with this acceleration, we find that sufficient sampling requires substantially longer simulations than are used in many published protocols. To further accelerate convergence, we introduce a new method of configuration space decomposition by orientation which reduces required simulation lengths by at least a factor of 5 in the cases examined. Our method is easily parallelizable, well suited for cases where a ligand cocrystal structure is not available, and can utilize initial orientations generated by docking packages.

Biophysics↗

Oncogene-dependent tumor suppression: using the dark side of the force for cancer therapy.

Cancers arise by an evolutionary process that involves the protracted acquisition by somatic cells of suites of interlocking mutations that uncouple proliferation, survival, migration, and damage responses from the mechanisms (selective pressures) that normally restrain or restrict them in time and space. The relative rareness of cancer cells within the soma, in the face of huge numbers of available cell targets, substantial rates of mutation, and an abundance of proto-oncogenes and tumor suppressor gene targets, indicates that the evolutionary space available to incipient tumor cells is highly restricted. The principal way in which this is achieved is through intrinsic tumor suppression pathways-innate growth arrest and apoptotic programs that fulfill an essentially analogous functional role to checkpoints in the cell cycle machinery by antagonizing the tumorigenic potential of oncogenic mutations. Using switchable transgenic and knockin mouse models, it is possible to identify these various tumor suppressor programs and establish where, when, how, and why they act to forestall neoplasia in each tissue type and, consequently, how and why their failure leads to cancer.

Animals↗

Feasibility of automated head-space gas chromatography in identification of anaerobic bacteria.

Neutral and acidic metabolites of some anaerobic bacteria cultured in a liquid growth medium were analysed by gas chromatography. Studies on the head-space vapours above heated broth cultures, using a gas chromatograph equipped with a fused silica capillary column and a unit for automatic head-space injection, enabled volatile alcohols and fatty acids to be readily detected. By contrast, when studying ether extracts of the same cultures, using a packed column, alcohols were only rarely registered, being more or less "hidden" under the solvent peak. Head-space gas chromatography was thus found to provide more diagnostic information on anaerobes, as exemplified by the registration of volatile alcohols produced by Clostridium sporogenes. Being suited for automation, this technique should be considered as an alternative to the study of solvent extracts of broth cultures in gas chromatographic identification of anaerobic bacteria.

Alcohols↗

Changes in cyst volume following intraoperative MRI-guided Ommaya reservoir placement for cystic craniopharyngioma.

INTRODUCTION: Intracavitary treatment of solitary cystic craniopharyngiomas with (32)P is an emerging treatment option, especially for pediatric patients. We have treated two patients with solitary cystic craniopharyngiomas using intraoperative MRI (iMRI)-guided catheter placement. METHODS: The optical tracking system of the General Electric Signa SP iMRI system was utilized for preoperative planning and intraoperative catheter tracking during insertion. Intraoperative volumetric imaging was then used to confirm final catheter position. Patients were brought back to the iMRI suite approximately 8 weeks later and diluted gadolinium was injected with further MRI to confirm the absence of communication between the cyst lumen and surrounding CSF spaces and for volumetric analysis. RESULTS: Intraoperative imaging illustrated deformation and changes in the cyst wall during catheter placement and cyst aspiration and confirmed final catheter placement. Images acquired 8 weeks following catheter placement prior to the instillation of (32)P showed decreases in cyst volume of 40 and 85%. CONCLUSION: iMRI-guided catheter placement for cystic craniopharyngiomas helps to assure successful catheter placement. Significant decreases in cyst volume occur in the interval between catheter placement and (32)P administration and must be accounted for to prevent overdosing of the radioisotope.

Adolescent↗

A new suite of programs for external beam radiotherapy isodose planning based on the RAD8 method.

A suite of programs is outlined which develops the ideas of the popular RAD-8-based approach to computerized RT treatment planning, but which uses real-space geometry throughout its algorithms. This, together with a higher level of modularity and complete documentation, enables other users of the software to understand the programs and adapt them to their special treatment methods. In addition, a number of new planning features are incorporated as useful options.

Computers↗

Changing documentation to improve stoma care.

Care pathways set out the anticipated flow of care for a typical patient with a specific diagnosis. The pathway of care is a concept rather than a fixed method, which can be adapted and modified to suit the needs of a specific patient group. Use of patient cards allows routine patient teaching and information-giving to be recorded, while allowing more space and time for focusing on the needs of individual patients.

Critical Pathways↗

[Hypobaric atmosphere and decompression safety of outer space explorations on the first days of flight].

54 subjects aged 21-47 participated in 8 series of experiments (298 altitude investigations) aimed at selecting decompression-safe protocols for transition from hypobaric normoxic environment to reduced pressure. Experimental data is discussed and some protocols for safe transition to reduced pressure are recommended. The experiments allowed to display in diagram form the ratio of permissible magnitudes of residual pressure (the working pressure in the extravehicular suit) to the duration of a preliminary exposure in normoxic hypobaric conditions.

Adaptation, Physiological↗

Cyclops: new modular software suite for cryo-EM.

Cyclops is a new computer program designed as a graphical front-end that allows easy control and interaction with tasks and programs for 3D reconstruction of biological complexes using cryo-electron microscopy. Cyclops' current plug-ins are designed for automated particle picking and include two new algorithms, automated carbon masking and quaternion based rotation space sampling, which are also presented here. Additional plug-ins are in the pipeline. Cyclops allows straightforward organization and visualization of all data and tasks and allows both interactive and batch-wise processing. Furthermore, it was designed for straightforward implementation in grid architectures. As a front-end to a collection of programs it provides a common interface to these programs, thus enhancing the usability of the suite and the productivity of the user.

Algorithms↗

Crystallization and preliminary X-ray diffraction studies of the Lb proteinase from foot-and-mouth disease virus.

Different crystal forms of the C23A mutant from the leader proteinase of foot-and-mouth disease virus were obtained by the hanging drop vapor diffusion technique, using MgCl2 and PEG 6000 as precipitants. Well-developed crystals, with cubic morphology growing to approximately 1.0 mm3 in size, presented a large unit cell parameter of 274.5 A and diffracted to, at most, 5 A resolution. A second type of crystal had a tetragonal appearance and these were obtained in droplets soaked in a silica gel matrix. These crystals, with an approximate size of 0.3 X 0.3 X 0.7 mm3, diffracted to approximately 4.0 A resolution, but presented a strong anisotropic mosaicity around the longest crystal axis. Crystals with a needlelike morphology and reaching sizes of about 0.2 X 0.3 X 1.2 mm3 diffracted beyond 3.5 A resolution and were stable to X-ray radiation for approximately one day when using a conventional source at room temperature. These crystals are orthorhombic with space group I222 (or I2(1)2(1)2(1)) and unit cell dimensions a = 65.9 A, b = 104.3 A, and c = 124.0 A, and appear well suited for high-resolution studies. Density packing considerations are consistent with the presence of two molecules in the asymmetric unit and a solvent content of approximately 54%.

Aphthovirus↗

Flexible ligand docking using a genetic algorithm.

Two computational techniques have been developed to explore the orientational and conformational space of a flexible ligand within an enzyme. Both methods use the Genetic Algorithm (GA) to generate conformationally flexible ligands in conjunction with algorithms from the DOCK suite of programs to characterize the receptor site. The methods are applied to three enzyme-ligand complexes: dihydrofolate reductase-methotrexate, thymidylate synthase-phenolpthalein and HIV protease-thioketal haloperidol. Conformations and orientations close to the crystallographically determined structures are obtained, as well as alternative structures with low energy. The potential for the GA method to screen a database of compounds is also examined. A collection of ligands is evaluated simultaneously, rather than docking the ligands individually into the enzyme.

Algorithms↗

Nucleotide sequence of the structural gene for diphtheria toxin carried by corynebacteriophage beta.

A 1,942-base-pair DNA segment encoding the structural gene for diphtheria toxin was sequenced, and the primary structure of the toxin was deduced. Restriction enzyme fragments corresponding to nontoxic or hypotoxic peptides of the toxin were isolated from corynebacteriophage beta and cloned into Escherichia coli on plasmid pBR322, and the sequence was determined. The mature toxin molecule deduced from the sequence has 535 amino acid residues and a molecular weight of 58,342. The deduced sequence for the fragment A moiety was the same as that determined at the protein level, except for a single serine residue, which had been mispositioned in the earlier study. Several differences were noted with respect to the partial sequence data available on the fragment B moiety, some or all of which may reflect genetic variations among populations of corynephages carrying the toxin gene. The DNA sequence predicts a 25-residue leader peptide preceding the mature protein, which is presumably involved in secretion of the toxin from lysogenized Corynebacterium diphtheriae. We infer that initiation of translation probably occurs at a GTG codon (codon -25). Cloned restriction fragments containing sequences for the amino-terminal region of toxin, together with 5' flanking regions, were expressed in E. coli. Toxin-related peptides were synthesized and secreted into the periplasmic space. These results provide a basis for applying recombinant DNA methods to the study of diphtheria toxin and for producing novel, genetically altered forms of the toxin suited to the construction of new classes of immunotoxins.

Amino Acid Sequence↗