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Genome size determination in peronosporales (Oomycota) by Feulgen image analysis.

Genome size was determined, by nuclear Feulgen staining and image analysis, in 46 accessions of 31 species of Peronosporales (Oomycota), including important plant pathogens such as Bremia lactucae, Plasmopara viticola, Pseudoperonospora cubensis, and Pseudoperonospora humuli. The 1C DNA contents ranged from 0.046 (45. 6 Mb) to 0.163 pg (159.9 Mb). This is 0.041- to 0.144-fold that of Glycine max (soybean, 1C = 1.134 pg), which was used as an internal standard for genome size determination. The linearity of Feulgen absorbance photometry method over this range was demonstrated by calibration of Aspergillus species (1C = 31-38 Mb) against Glycine, which revealed differences of less than 6% compared to the published CHEF data. The low coefficients of variation (usually between 5 and 10%), repeatability of the results, and compatibility with CHEF data prove the resolution power of Feulgen image analysis. The applicability and limitations of Feulgen photometry are discussed in relation to other methods of genome size determination (CHEF gel electrophoresis, reassociation kinetics, genomic reconstruction) that have been previously applied to Oomycota.

Aspergillus↗

Is H2O present on Io? The detection of a new strong band near 3590 cm-1 (2.79 micrometers).

A strong absorption band at 3590 +/- 20 cm-1 (2.790 +/- 0.015 micrometers) has been discovered in the spectrum of Io using the Kuiper Airborne Observatory (KAO). The 2 nu1 + nu3 combination mode of solid SO2 falls at this position. Since SO2 is abundant on Io it must contribute to the new band. However, a band due to H2O was predicted near this frequency in Io's spectrum based on laboratory experiments of H2O:SO2 mixed Io ice analogs which were used to assign the two weak, variable features of 3370 and 3170 cm-1 (2.97 and 3.15 micrometers) to trace amounts of H2O frozen in solid SO2 on Io. The new band probably originates from both SO2 and H2O. Unfortunately, the spectral resolution of the data is insufficient to settle the issue of whether there are two resolvable components.

Aircraft↗

Enamel thickness and the topography of the enamel-dentine junction in South African Plio-Pleistocene hominids with special reference to the Carabelli trait.

This study explores the internal morphology of early hominid teeth using high-resolution computed tomography. Data on Carabelli feature size, enamel thickness, and the topography of the enamel-dentine junction are considered together in order to examine the relationship among these variables in the maxillary molars of gracile and robust australopithecines from South Africa. In particular, one aim is to investigate the degree to which Carabelli feature size influences enamel thickness in the plane of the mesial cusps. The results demonstrate that maxillary molars attributed to Australopithecus africanus from Sterkfontein, Taung and Makapansgat possess larger Carabelli features and thinner enamel along the lingual wall of the protocone than do specimens attributed to Paranthropus robustus from Swartkrans and Kromdraai. Distinct differences in the position of the Carabelli feature at the level of both the enamel-dentine junction and tooth crown surface between early hominid species may help explain the observed disparity in enamel thickness at that region of the tooth crown as well as offer clues to the functional role of Carabelli's cusp. As the size and position of the Carabelli feature affects the linear thickness of enamel at this one particular region of the tooth crown, future comparative studies focusing on taxa that possess moderate to strong development of the Carabelli complex should use the linear thickness of enamel taken close to the protoconal dentine horn or at the maximum projection of the Carabelli's cusp.

Animals↗

Satisfying hydrogen bonding potential in proteins.

We have analysed the frequency with which potential hydrogen bond donors and acceptors are satisfied in protein molecules. There are a small percentage of nitrogen or oxygen atoms that do not form hydrogen bonds with either solvent or protein atoms, when standard criteria are used. For high resolution structures 9.5% and 5.1% of buried main-chain nitrogen and oxygen atoms, respectively, fail to hydrogen bond under our standard criteria, representing 5.8% and 2.1% of all main-chain nitrogen and oxygen atoms. We find that as the resolution of the data improves, the percentages fall. If the hydrogen bond criteria are relaxed many of these unsatisfied atoms form weak hydrogen bonds. However, there remain some buried atoms (1.3% NH and 1.8% CO) that fail to hydrogen bond without any immediately obvious compensating interactions.

Algorithms↗

Actin: from cell biology to atomic detail.

Over the past 2 decades our knowledge about actin filaments has evolved from a rigid "pearls on a string" model to that of a complex, highly dynamic protein polymer which can now be analyzed at atomic detail. To achieve this, exploring actin's oligomerization, polymerization, polymorphism, and dynamic behavior has been crucial to understanding in detail how this abundant and ubiquitous protein can fulfill its various functions within living cells. In this review, a correlative view of a number of distinct aspects of actin is presented, and the functional implications of recent structural, biochemical, and mechanical data are critically evaluated. Rational analysis of these various experimental data is achieved using an integrated structural approach which combines intermediate-resolution electron microscopy-based 3-D reconstructions of entire actin filaments with atomic resolution X-ray data of monomeric and polymeric actin.

Actin Cytoskeleton↗

Crystallization and preliminary X-ray crystallographic analysis of spruce budworm antifreeze protein.

Antifreeze proteins have the ability to bind to ice with high affinity and inhibit further crystal growth. The insect antifreeze protein from spruce budworm exhibits very high thermal hysteresis activity and is implicated in the protection of overwintering larvae from freezing. This protein has been crystallized in 20-25% polyethylene glycol (Mr 6000), 0.4 M NaCl, 0.1 M Tris-HCl, pH 8.5, by vapor diffusion using the hanging drop method. The resulting crystals are very thin (typically <0.01 mm in the shortest dimension), and only after repeated seeding could crystals be grown large enough for data collection using synchrotron radiation. The crystals belong to the monoclinic space group C2, with cell dimensions a = 82.28 A, b = 62.29 A, c = 63.63 A, and beta = 113.7 degrees. Molecules in the asymmetric unit are related by a twofold axis of symmetry with two molecules present. Native data to a resolution of 2.6 A have been collected with 90.3% completeness and a Rsym of 6.9%.

Animals↗

Microdialysis coupled to online enzymatic assays.

In vivo sampling of interstitial fluid by using microdialysis fibers has become a standard and accepted procedure. This sampling method is generally coupled to offline analysis of consecutive dialysate samples by high-performance liquid chromatography or capillary electrophoresis, but this combination is not the best approach for some applications, especially those which require high temporal resolution and rapid data collection. The purpose of this review is to provide information on enzyme-based online assays, i.e., continuous analysis of the dialysate as it emerges from the outlet of the sampling device. We have focused on methods developed specifically for the analysis of solutions perfused at a very slow flow rate, i.e., a feature of microdialysis and ultrafiltration techniques. These methods include flow enzyme-fluorescence assays, flow enzyme-amperometric assays, and sequential enzyme-amperometric detection. Each type of assay is discussed in terms of principle, applications, advantages, and limitations. We also comment on implantable biosensors, an obvious next step forward for in vivo monitoring of molecules in neuroscience.

Animals↗

Molecular insights into species phylogeny, biogeography, and morphological stasis in the ancient spider genus Hypochilus (Araneae: Hypochilidae).

The spider genus Hypochilus is currently restricted to cool, moist microhabitats in three widely separated montane regions of North America, providing an opportunity to study both deep (i.e., continental level) and shallow (within montane region) biogeographic history. Members of the genus also retain many plesiomorphic morphological characteristics, inviting the study of comparative rates of morphological evolution. In this paper, Hypochilus phylogeny and associated evolutionary problems are addressed using both new molecular (28S nDNA and CO1 mtDNA) and previously published (K. M. Catley, 1994, Am. Mus. Nov. 3088, 1-27) morphological data. Although the molecular data provide limited resolution of root placement within Hypochilus, most analyses are at least consistent with morphology-supported montane relationships of (Rockies (California, Appalachian)). The monophyly of Hypochilus species distributed in the California mountains is ambiguous, with several analyses indicating that this fauna may be paraphyletic with respect to a monophyletic Appalachian lineage. The montane regions differ in consistent ways in depths of both mitochondrial and nuclear phylogenetic divergence. Molecular clock analyses, in combination with arthropod-based mtDNA rate calibrations, suggest that the regional faunas are of different ages and that speciation in all faunas likely occurred prior to the Pleistocene. Limited intraspecific sampling reveals extraordinarily high levels of mtDNA cytochrome oxidase sequence divergence. These extreme divergences are most consistent with morphological stasis at the species level, despite preliminary evidence that Hypochilus taxa are characterized by fragmented population structures.

Animals↗

DNA cloning in Lactobacillus helveticus by the exconjugation of recombinant mob-containing plasmid constructs from strains of transformable lactic acid bacteria.

A system developed for the genetic transfer of plasmids between strains of nontransformable bacteria (P. Langella, Y. le Loir, S. D. Ehrlich and A. Gruss, 1993, J. Bacteriol., 175, 5806-5813) by the specific inclusion of a mobilization (mob) region into a nonconjugative shuttle vector was used successfully to deliver the genetic determinants for beta-glucanase, beta-glucuronidase, and green fluorescent protein to Lactobacillus helveticus. Expression of two of the genes could be detected in the new host. Data suggested that resolution of cointegrates into components could release the original recombinant plasmid or generate a cointegrate deletion. All the recombinant plasmids were segregationally unstable in Lb. helveticus and there was some evidence for structural instability. Intrinsic instability in the mob-containing vector was reduced by replacing the duplicated pBluescript polylinker with that from pUC19. Sites at which cointegrate formation could occur were localized at two distinct tracts close to the D-loop that forms at the primosome during plasmid replication.

Bacterial Proteins↗

NMR and time-resolved optical studies of brain imaging.

The classical optical method of the biochemist used in so many applications to determine stoichiometry kinetics, quantitative analysis of biochemical systems which was subsequently applied to tissue studies by G.A. Millikan and others seems now to have a new lease on life provided by time and frequency resolved studies which appear to afford the essential quantification lacking in the earlier methods. The fact that localization seems possible as well may bring the optical technology into the arena of new non-invasive technology capable of high sensitivity, high resolution and rapid data acquisition.

Animals↗

Genus zero surface conformal mapping and its application to brain surface mapping.

It is well known that any genus zero surface can be mapped conformally onto the sphere and any local portion thereof onto a disk. However, it is not trivial to find a general method which finds a conformal mapping between two general genus zero surfaces. We propose a new variational method which can find a unique mapping between any two genus zero manifolds by minimizing the harmonic energy of the map. We demonstrate the feasibility of our algorithm by applying it to the cortical surface matching problem. We use a mesh structure to represent the brain surface. Further constraints are added to ensure that the conformal map is unique. Empirical tests on MRI data show that the mappings preserve angular relationships, are stable in MRIs acquired at different times, and are robust to differences in data triangulation, and resolution. Compared with other brain surface conformal mapping algorithms, our algorithm is more stable and has good extensibility.

Algorithms↗

Magnetic resonance: a multimodal approach to the brain?

Magnetic resonance of the brain offers the unique possibility of acquisition of morphological data in high resolution quality, analysis of metabolic disturbances, and the assessment of brain function. Basic principle is the BOLD effect, using the difference of the magnetic moments of oxy- and deoxyhemoglobin as intrinsic mechanism of contrast. Still the optimum techniques for functional imaging are not established: echo planar imaging requires a very costly hardware, whereas gradient echo imaging is feasible on clinical routine MR scanners. Although many experimental studies have been published on visual, motor, and auditory paradigms, clinical applications have not been established up to now. Analysis of epileptogenic foci combining high resolution imaging and functional information seems very promising as well as preoperative determination of critical areas in neurosurgery.

Brain↗

The superstructure of chromatin and its condensation mechanism. III: Effect of monovalent and divalent cations X-ray solution scattering and hydrodynamic studies.

Solutions of rat liver and chicken erythrocyte chromatin at different ionic strengths were characterized by synchrotron X-ray solution scattering, ultracentrifugation, density and viscosity measurements. Previous observations on nuclei were extended to rat liver, calf thymus and yeast nuclei. It is shown that with monovalent cations condensation is independent of the nature of the cation whereas with divalent cations there are significant differences related to the preference of base binding over phosphate binding. The consistency of hydrodynamic and scattering results confirm the view that chromatin in solution at low ionic strength has a helix-like superstructure. A survey of X-ray and neutron scattering results in the literature shows that previous interpretations, e.g. in terms of a 10 nm filament, are incompatible with the experimental data at low resolution.

Animals↗

Dynamic properties of cat tenuissimus muscle.

Some of the factors which influence the development of tension in cat tenuissimus muscle were studied quantitatively. Under isometric conditions, it was shown that the dynamic properties of the relationship between the tension of the muscle and its electrical stimulation depend on the mean rate of stimulation. This non-linear effect cannot be explained on the basis of the dependence of muscle tension on instantaneous rate of stimulation since the tension due to a stimulus following closely a previous stimulus is augmented, but the time course of the twitch response is unaltered. The interaction between the tension due to active contraction and that due to the viscoelastic properties of the muscle was investigated by independently varying muscle length and the rate of stimulation. Within the limits of resolution of the data, it was concluded that these two components of tension are addictive and that muscle stiffness is related to the instantaneous tension of the muscle.

Animals↗

Computer assisted brain surgery for small lesions in the central sensorimotor region.

The capacity of a new optical navigation device is demonstrated by six microsurgical procedures for small subcortical lesions within the central sensorimotor strip. This small series is aimed at less invasive resection in this functionally critical region, independently of primary diagnosis and outcome. Guided by high resolution CT imaging data five brain tumours and one cavernous angioma was selectively located and most sparingly removed without additional sensorimotor deficit. In two cases improvement of a pre-operative paresis was observed immediately after surgery. Thanks to light-weight freehand pointing instruments and a ranging accuracy of +/- 1 mm, damage to functionally important brain areas and vessels was avoided by using uncommonly oblique, e.g., transsulcal ways of access which would hardly have been possible even with guidance by conventional stereotaxy. The demanding systematic cortical stimulation of the precentral gyrus applied in three cases was only sensitive in infiltrating tumours-e.g., low grade astrocytomas-where for want of adjuvant therapy it was essential to proceed to the extreme limits of resection. In general, precise anatomical localisation by computer aided surgery (CAS) is sufficient in small central lesions which guarantees minimally invasive surgery. The potential of this new, soon commercially available optical navigation system in (neuro) surgery, quality control and teaching is discussed.

Adult↗

A desktop magnetic resonance imaging system.

Modern magnetic resonance imaging (MRI) systems consist of several complex, high cost subsystems. The cost and complexity of these systems often makes them impractical for use as routine laboratory instruments, limiting their use to hospitals and dedicated laboratories. However, advances in the consumer electronics industry have led to the widespread availability of inexpensive radio-frequency integrated circuits with exceptional abilities. We have developed a small, low-cost MR system derived from these new components. When combined with inexpensive desktop magnets, this type of MR scanner has the promise of becoming standard laboratory equipment for both research and education. This paper describes the development of a prototype desktop MR scanner utilizing a 0.21 T permanent magnet with an imaging region of approximately 2 cm diameter. The system uses commercially available components where possible and is programmed in LabVIEW software. Results from 3D data sets of resolution phantoms and fixed, newborn mice demonstrate the capability of this system to obtain useful images from a system constructed for approximately $13,500.

Animals↗

[Intraoperative three-dimensional navigation for pedicle screw placement].

The mobile SIREMOBIL Iso-C(3D) C-arm (Siemens, Erlangen, Germany) is the first device permitting intraoperative, three-dimensional representation of bone structures. A high-resolution, isotropic 3D data cube in the isocenter with sides of approximately 12 cm is calculated simultaneously. The SIREMOBIL Iso-C(3D) is linked to the navigation system. This makes it possible to transfer the generated 3D data directly to the linked navigation system without the need for surgeon-dependent registration. In this prospective clinical trial, we evaluated the accuracy of pedicle screw placement using this device. In 61 patients, a total of 302 pedicle screws were placed. Only in five cases (1.7%) were misplacements of > or =2 mm shown in postoperative control CT. The average fluoroscopy time was 1.28+/-0.56 min, and the average operative duration was 103.26+/-23.3 min. There were no postoperative neurological complications in any of the 30 patients. From these data, we conclude that Iso-C(3D) navigation is a very accurate method for the placement of pedicle screws.

Adolescent↗

[Iso-C(3D0-assisted) navigated implantation of pedicle screws in thoracic lumbar vertebrae].

The mobile Siremobil Iso-C(3D) C-arm (Siemens AG, Medical Solutions, Erlangen) is the first device that permits the intraoperative three-dimensional (3D) representation of bone structures. A high-resolution isotropic 3D data cube in the isocenter with an edge length of approximately 12 cm is calculated simultaneously. The Siremobil Iso-C(3D) is linked to navigation with the integrated NaviLink interface (Siemens AG, Medical Solutions, Erlangen). This makes it possible to transfer the generated 3D data directly to the linked navigation system Surgigate (Medivision, Oberndorf, Switzerland). In this prospective clinical trial we evaluated the accuracy of pedicle screw placement using the Siremobil Iso-C(3D) C-arm. The results were compared to the conventional approach and other computer-assisted procedures (CT-based navigation, C-arm-based 2D navigation) in historical control groups. A total of 141 pedicle screws were placed in 30 patients (70 thoracic spine, 71 lumbar spine). Only in one single case was misplacement shown in the postoperative control CT scan (0.71%), the lowest rate of incorrect placements of all techniques. Also the lowest average fluoroscopy time (1.28+/-0.56 min) was achieved during the placement of pedicle screws on the spine with Iso-C(3D) navigation at a comparable average OR duration (103.26+/-23.3 min). There were no postoperative neurological complications in all 30 patients. From these data we conclude that Iso-C(3D) navigation of pedicle screws is a very accurate method in the correct placement of pedicle screws.

Bone Screws↗