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Calcium signalling--an overview.

Calcium (Ca2+) is an almost universal intracellular messenger, controlling a diverse range of cellular processes, such as gene transcription, muscle contraction and cell proliferation. The ability of a simple ion such as Ca2+ to play a pivotal role in cell biology results from the facility that cells have to shape Ca2+ signals in the dimensions of space, time and amplitude. To generate the variety of observed Ca2+ signals, different cell types employ components selected from a Ca2+ signalling 'toolkit', which comprizes an array of signalling, homeostatic and sensory mechanisms. By mixing and matching components from the toolkit, cells can obtain Ca2+ signals that suit their physiology.

Animals↗

[MRT of the eye: the normal anatomy and detection of the smallest lesions with a high-resolution surface coil].

PURPOSE: A new, high-resolution surface coil for MRI of the eye was evaluated with regard to practicability, image quality and sensitivity for small lesions. MATERIAL AND METHODS: 48 patients in whom a space-occupying lesion of the eye or orbit was suspected were examined (1.5 T tomograph, 5 cm surface coil, T1- and T2-weighted spin-echo sequences, the former before and after i.v. gadolinium DTPA). RESULTS: 45/48 patients tolerated MR with the high-resolution surface coil. No adverse effects were experienced by the patients. In 11/48 patients a space occupying lesion of the eye was detected (melanoma, 5; metastases, 2; haemorrhage, 1; malformation, hamartoma and scarring after melanoma, one each). The smallest detectable lesion had a thickness of < 1 mm. CONCLUSION: First experiences with the high-resolution surface coil indicate that this device is suited for detection of very small lesions of the eye.

Adolescent↗

Hypobaric biology: Arabidopsis gene expression at low atmospheric pressure.

As a step in developing an understanding of plant adaptation to low atmospheric pressures, we have identified genes central to the initial response of Arabidopsis to hypobaria. Exposure of plants to an atmosphere of 10 kPa compared with the sea-level pressure of 101 kPa resulted in the significant differential expression of more than 200 genes between the two treatments. Less than one-half of the genes induced by hypobaria are similarly affected by hypoxia, suggesting that response to hypobaria is unique and is more complex than an adaptation to the reduced partial pressure of oxygen inherent to hypobaric environments. In addition, the suites of genes induced by hypobaria confirm that water movement is a paramount issue at low atmospheric pressures, because many of gene products intersect abscisic acid-related, drought-induced pathways. A motivational constituent of these experiments is the need to address the National Aeronautics and Space Administration's plans to include plants as integral components of advanced life support systems. The design of bioregenerative life support systems seeks to maximize productivity within structures engineered to minimize mass and resource consumption. Currently, there are severe limitations to producing Earth-orbital, lunar, or Martian plant growth facilities that contain Earth-normal atmospheric pressures within light, transparent structures. However, some engineering limitations can be offset by growing plants in reduced atmospheric pressures. Characterization of the hypobaric response can therefore provide data to guide systems engineering development for bioregenerative life support, as well as lead to fundamental insights into aspects of desiccation metabolism and the means by which plants monitor water relations.

Adaptation, Physiological↗

Scoring noncovalent protein-ligand interactions: a continuous differentiable function tuned to compute binding affinities.

Exploitation of protein structures for potential drug leads by molecular docking is critically dependent on methods for scoring putative protein-ligand interactions. An ideal function for scoring must exhibit predictive accuracy and high computational speed, and must be tolerant of variations in the relative protein-ligand molecular alignment and conformation. This paper describes the development of an empirically derived scoring function, based on the binding affinities of protein-ligand complexes coupled with their crystallographically determined structures. The function's primary terms involve hydrophobic and polar complementarity, with additional terms for entropic and solvation effects. The issue of alignment/conformation dependence was solved by constructing a continuous differentiable nonlinear function with the requirement that maxima in ligand conformation/alignment space corresponded closely to crystallographically determined structures. The expected error in the predicted affinity based on cross-validation was 1.0 log unit. The function is sufficiently fast and accurate to serve as the objective function of a molecular-docking search engine. The function is particularly well suited to the docking problem, since it has spatially narrow maxima that are broadly accessible via gradient descent.

Bacterial Proteins↗

Asymmetrical masking between radial and parallel motion flow in transparent displays.

Human spatial behavior remains surprisingly accurate and stable, despite the complexities of the concurrent retinal image flow. How does visual motion analysis contribute to behavioral stability? To approach this question, we rely on current knowledge about heading perception and coherent motion extraction from animated random dot displays. This chapter presents psychophysical evidence for an asymmetry in the global visual decomposition of the retinal motion flow. We examined the ability of human observers to identify the radial and the parallel flow component in displays where the two motions were transparently superimposed. Whereas sensitivity to the radial motion remained unaffected by the presence of parallel flow, sensitivity to the latter was greatly degraded in the presence of radial flow. This imbalance was observed within the first 200 ms of motion exposure, prior to the perception of heading (430 ms; Hooge et al. (1999) Exp. Brain Res., 129: 615-628). Although such an asymmetrical masking is well suited to minimize heading errors, whether it really contributes to the extraction of behavioral direction remains an open question.

Animals↗

A recirculatory model of the pulmonary uptake and pharmacokinetics of lidocaine based on analysis of arterial and mixed venous data from dogs.

Pulmonary uptake of basic amine xenobiotics such as lidocaine may influence the onset of drug effect and ameliorate toxicity. To date, pharmacokinetic analysis of pulmonary drug uptake has been only semiquantitative and ill-suited for relating pharmacodynamics to pharmacokinetics or for estimating the time course of the fraction of drug dose residing in the lung during a single pass. We have developed recirculatory models in an experiment in which lidocaine was injected into the right atrium simultaneously with markers of intravascular space (indocyanine green) and total body water (antipyrine); this was followed by rapid arterial and mixed venous blood sampling. Such models are interpretable physiologically and are capable of characterizing the kinetics of the pulmonary uptake of lidocaine in addition to peripheral tissue distribution and elimination. The apparent pulmonary tissue volume of lidocaine (39 ml/kg) was nearly ninefold greater than that of antipyrine (4.5 ml/kg). The recirculatory model characterized both arterial and mixed venous data, but the latter data were not essential for estimating lidocaine's pulmonary disposition either before or after recirculation of drug was evident.

Anesthetics, Local↗

A state-specific approach to multireference coupled electron-pair approximation like methods: development and applications.

The traditional multireference (MR) coupled-cluster (CC) methods based on the effective Hamiltonian are often beset by the problem of intruder states, and are not suitable for studying potential energy surface (PES) involving real or avoided curve crossing. State-specific MR-based approaches obviate this limitation. The state-specific MRCC (SS-MRCC) method developed some years ago can handle quasidegeneracy of varying degrees over a wide range of PES, including regions of real or avoided curve-crossing. Motivated by its success, we have suggested and explored in this paper a suite of physically motivated coupled electron-pair approximations (SS-MRCEPA) like methods, which are designed to capture the essential strength of the parent SS-MRCC method without significant sacrificing its accuracy. These SS-MRCEPA theories, like their CC counterparts, are based on complete active space, treat all the reference functions on the same footing and provide a description of potentially uniform precision of PES of states with varying MR character. The combining coefficients of the reference functions are self-consistently determined along with the cluster amplitudes themselves. The newly developed SS-MRCEPA methods are size-extensive, and are also size-consistent with localized orbitals. Among the various versions, there are two which are invariant with respect to the restricted rotations among doubly occupied and active orbitals separately. Similarity of performance of this latter and the noninvariant versions at the crossing points of the degenerate orbitals imply that the all the methods presented are rather robust with respect to the rotations among degenerate orbitals. Illustrative numerical applications are presented for PES of the ground state of a number of difficult test cases such as the model H4, H(8) problems, the insertion of Be into H(2), and Li(2), where intruders exist and for a state of a molecule such as CH(2), with pronounced MR character. Results obtained with SS-MRCEPA methods are found to be comparable in accuracy to the parent SS-MRCC and FCI/large scale CI results throughout the PES, which indicates the efficacy of our SS-MRCEPA methods over a wide range of geometries, despite their neglect of a host of complicated nonlinear terms, even when the traditional MR-based methods based on effective Hamiltonians fail due to intruders.

Journal Article↗

Evidence for TeV Emission from GRB 970417a.

Milagrito, a detector sensitive to very high energy gamma rays, monitored the northern sky from 1997 February through 1998 May. With a large field of view and a high duty cycle, this instrument was well suited to perform a search for TeV gamma-ray bursts (GRBs). We report on a search made for TeV counterparts to GRBs observed by BATSE. BATSE detected 54 GRBs within the field of view of Milagrito during this period. An excess of events coincident in time and space with one of these bursts, GRB 970417a, was observed by Milagrito. The excess has a chance probability of 2.8x10-5 of being a fluctuation of the background. The probability for observing an excess at least this large from any of the 54 bursts is 1.5x10-3. No significant correlations were detected from the other bursts.

Journal Article↗

Reconstruction of calvarial defects.

We subdivide the calvarium into three zones, each with its special reconstructive requirements. Based on our experience with calvarial defects in 13 patients, we favor use of autogenous material, especially in the face of previous infection or a scarred recipient bed. Alloplasts give excellent forehead contour but alloplastic reconstruction should be delayed for 1 year after injury. Vascularized bone grafts maintain contour well. They are best suited to large periorbital defects. At other locations we favor split calvarial free bone grafts. Occasionally, the defect may be so large as to warrant grafts from multiple donor sites. Use of vascularized muscle helps eradicate infection, provides a vascularized bed for free bone grafts, and fills dead space. The frontal sinus is managed either by cranialization (if the posterior wall is involved) or by mucosal stripping with obliteration of the nasofrontal duct. Additional technical considerations include rigid bone fixation, surgical exposure through a bicoronal incision, and meticulous handling of bone grafts.

Adolescent↗

Continuous distributions of ventilation-perfusion ratios in normal subjects breathing air and 100 per cent O2.

A new method has been developed for measuring virtually continuous distributions of ventilation-perfusion ratios (V(A)/Q) based on the steadystate elimination of six gases of different solubilities. The method is applied here to 12 normal subjects, aged 21-60. In nine, the distributions were compared breathing air and 100% oxygen, while in the remaining three, effects of changes in posture were examined. In four young semirecumbent subjects (ages 21-24) the distributions of blood flow and ventilation with respect to V(A)/Q were virtually log-normal with little dispersion (mean log standard deviations 0.43 and 0.35, respectively). The 95.5% range of both blood flow and ventilation was from V(A)/Q ratios of 0.3-2.1, and there was no intrapulmonary shunt (V(A)/Q of 0). On breathing oxygen, a shunt developed in three of these subjects, the mean value being 0.5% of the cardiac output. The five older subjects (ages 39-60) had broader distributions (mean log standard deviations, 0.76 and 0.44) containing areas with V(A)/Q ratios in the range 0.01-0.1 in three subjects. As for the young subjects, there was no shunt breathing air, but all five developed a shunt breathing oxygen (mean value 3.2%), and in one the value was 10.7%. Postural changes were generally those expected from the known effects of gravity, with more ventilation to high V(A)/Q areas when the subjects were erect than supine. Measurements of the shunt while breathing oxygen, the Bohr CO(2) dead space, and the alveolar-arterial oxygen difference were all consistent with the observed distributions. Since the method involves only a short infusion of dissolved inert gases, sampling of arterial blood and expired gas, and measurement of cardiac output and minute ventilation, we conclude that it is well suited to the investigation of pulmonary gas exchange in man.

Acetone↗

Effects of chemotherapy by 1,3-bis(2-chloroethyl)-1-nitrosourea on single-quantum- and triple-quantum-filtered 23Na and 31P nuclear magnetic resonance of the subcutaneously implanted 9L glioma.

The effects of chemotherapy [25 mg/kg 1,3-bis(2-chloroethyl)-1-nitrosourea administered with a single i.p. injection] on cellular energetics by 31P nuclear magnetic resonance (NMR) spectroscopy, total tissue sodium by single-quantum (SQ) 23Na NMR spectroscopy, and intracellular sodium by triple-quantum-filtered (TQF) 23Na NMR spectroscopy were studied in the s.c. 9L glioma. Animals were studied by NMR 2 days before therapy and 1 and 5 days after therapy. Destructive chemical analysis was also performed 5 days after therapy to validate the origin of changes in SQ and TQF 23Na signals. One day after treatment, there was no significant difference between control and treated tumors in terms of tumor size or 23Na and 31P spectral data. Five days after therapy, treated tumors had 28 +/- 16% (P < 0.1) lower SQ 23Na signal intensity, 46 +/- 20% (P < 0.05) lower TQF 23Na signal intensity, 125 +/- 51% (P < 0.05) higher ATP:Pi ratio, 186 +/- 69% (P < 0.05) higher phosphocreatine:Pi ratio, and 0.17 +/- 0.06 pH units (P < 0.05) higher intracellular pH compared with control tumors. No significant differences in TQF 23Na relaxation times were seen between control and treated tumors at any time point. Destructive chemical analysis showed that the relative extracellular space of control and treated tumors was identical, but the treated tumors had 21 +/- 8% (P < 0.05) lower total tissue Na+ concentration and 60 +/- 24% (P < 0.05) lower intracellular Na+ concentration compared with the controls. The higher phosphocreatine:Pi and ATP:Pi ratios after 1,3-bis(2-chloroethyl)-1-nitrosourea treatment indicate improved bioenergetic status in the surviving tumor cells. The decrease in SQ and multiple-quantum-filtered 23Na signal intensity was largely attributable to a decrease in Na(i)+ because the treatment did not change the relative extracellular space. The improved energy metabolism could decrease the intracellular concentration of Na+ by increasing the activity of Na+-K+-ATPase and decreasing the activity of Na+/H+. Although both 23Na and 31P spectra were consistent with improved cellular metabolism in treated tumors, the 23Na methods may be better suited for monitoring response to therapy because of higher signal:noise ratio and ease of imaging the single 23Na resonance.

Adenosine Triphosphate↗

The medical legacy of Gemini.

The Mercury and Gemini space flights have provided approximately 2,000 manhours of weightless exposure which can be used in comparing flight results with the predicted effects of manned space flight. In general the environmental hazards and the effects upon man appear to be of less magnitude than originally anticipated. The effects noted on the various body systems are summarized. The principal physiologic changes noted were orthostatism for some 50 hours post-flight, reduced red cell mass, and reduced X-ray density in the os calcis and the small finger. Much was learned about man's ability to work in a pressurized suit in the extravehicular condition. Early biochemical findings have pointed the way to future investigations. All of these findings are of importance in relation to the planning for future long duration missions. Although much remains to be learned, it does appear from an overview of this medical legacy that if man is properly supported and evaluated his limitations will not be a barrier to the exploration of the universe.

Adaptation, Physiological↗

Characterisation of a novel solid lipid nanoparticle carrier system based on binary mixtures of liquid and solid lipids.

A drug carrier of colloidal lipid particles with improved payloads and enhanced storage stability was investigated. Based on the experiences with hard fats nanoparticles, a new type of solid lipid nanoparticles (SLN) has been developed by incorporating triglyceride containing oils in the solid core of said particle. The structure and mixing behaviour of these particles were characterised and practical implications on controlled release properties tested. Nanoparticles were characterised by their melting and recrystallisation behaviour as recorded by differential scanning calorimetry (DSC). Polymorphic form and bilayer arrangement were assigned by wide-angle X-ray scattering (WAXS) and small-angle X-ray scattering (SAXS). Size distribution and storage stability were investigated by laser diffractometry (LD). Release properties were studied by drug release model according to Franz. A medium chain triglyceride oil was incorporated successfully in a matrix of a solid long chain glyceride. The crystal order was greatly disturbed, however, the carrier remained solid. The oil inside the particle remained in a liquid state and induced a slight shift form the beta' polymorph to the beta(i) form. Long spacings varied within 0.1 nm with increasing oil loads. Nanoparticles with low oil concentrations showed sustained release properties. Improved drug load levels were encapsulated by lipid particles supplemented with oily constituents. Thus, the presented carrier adds additional benefits to the well-known opportunities of conventional SLN and is suited for topical use.

Calorimetry, Differential Scanning↗

Posthemorrhagic ventricular dilation in the neonate: development and characterization of a rat model.

Intraventricular hemorrhage is a common complication of prematurity. Posthemorrhagic ventricular dilation (PHVD) has a high rate of disability and no safe and effective treatment. Its pathogenesis is poorly understood, largely because of the lack of a satisfactory animal model. We have developed a model of neonatal PHVD in the rat. Seven-day-old (P7) Wistar rat pups were given 80-microl injections of citrated rat blood or artificial cerebrospinal fluid (CSF) into alternate lateral ventricles on P7 and P8. Intracranial pressure was monitored and increased briefly by over 8-fold. Some rats received further 10-microl intraventricular injections of India ink on P21. Animals were weighed daily and simple neurologic tests performed. On P21 (or P22 if India ink had been injected), the rats were perfusion-fixed and blocks processed for paraffin histology. Sixty-five percent of pups injected with blood and 50% injected with artificial CSF developed dilated lateral ventricles, with patchy loss of ependyma, marked astrocytic gliosis, and rarefaction of periventricular white matter. India ink injection revealed slow transit of CSF from the dilated lateral ventricles but eventual passage into the subarachnoid space. Pups that had received intraventricular injections but did not develop ventricular dilation nonetheless had lighter brains than littermate controls (p < 0.001). Body weights were not significantly different from controls. Hydrocephalic animals had reduced motor performance as assessed by a grip traction test (p = 0.0002). This model is well suited to studying the pathogenesis of PHVD.

Animals↗

Ultrafast pulse sequence techniques for cardiac magnetic resonance imaging.

Cardiac magnetic resonance imaging is a rapidly emerging field that has seen tremendous advances in the past decade. Central to the development of effective imaging strategies has been the advent of high-performance gradient hardware and the exploitation of their speed characteristics through specialized pulse sequences well suited for cardiac imaging. These advances have facilitated unprecedented acquisition times that now approach echocardiographic frame rates, while maintaining excellent image quality. This article provides a detailed overview of advanced pulse sequence technology and approaches currently taken to maximize speed performance and image quality. In particular, segmented K-space techniques that include single-echo and multiecho spoiled gradient-echo imaging as well as steady-state free precession imaging are discussed. Finally, spiral and fast spin-echo techniques are explored. Examples of common applications of these pulse sequences are presented.

Echo-Planar Imaging↗

Interactive magnetic resonance imaging-guided biopsy in the head and neck: initial patient experience.

Because of its excellent soft tissue resolution, magnetic resonance imaging (MRI) can optimize image guidance for interventional and surgical procedures. Notably, needle biopsy of head and neck lesions has been used for years, deeper lesions often requiring some form of image guidance. The closed space of diagnostic MRI scanners proves cumbersome for intervention. The authors report on the first head and neck image-guided biopsies performed in a new, investigational "open configuration" intraoperative MRI scanner. Vertical space between the scanner's upright coils gives access to the patient while imaging; image acquisition is as fast as 2 sec/image. Biopsies were performed on seven patients (parotid, parapharyngeal space, second cervical vertebra); five specimens were diagnostic. Both general anesthesia and intravenous sedation were used. The procedures were without complications. Imaging provided definition of anatomy to direct needle placement. Access to the patient allowed for both percutaneous and transoral approaches. The environment of the open magnet is well suited for biopsy of the head and neck, and near real-time intraoperative MRI has promise for guiding more complex head and neck procedures. Further study should optimize the quality of the images and the interactibility of localization and targeting and fully utilize MRI's three-dimensional imaging capabilities.

Adenoma, Pleomorphic↗

An analysis of neural models for walking control.

A large space of different neural models exists from simple mathematical abstractions to detailed biophysical representations with strongly differing levels of complexity and biological relevance. Previous comparisons between models have looked at biological realism or mathematical tractability rather than expressive power. This paper, however, investigates whether more sophisticated models are better suited to a complex sensorimotor control task than simpler ones, or whether the more general nature of groups of the simpler neurons allows them to collectively solve complex tasks better despite their individual simplicity. Many models have been proposed or used for sensorimotor control tasks such as the control of locomotion. Four such neural models with varying levels of complexity were chosen. Controllers made of networks of each neural type were evolved to generate locomotion in a simulated dynamically stable four-legged robot using a genetic algorithm. The problem domain was chosen as one for which no simple solution could be hand crafted and which, with its tight sensorimotor coupling, had strongly time-dependent properties as is common in many biological control tasks. Analysis of the results shows that the most complex and biologically based model is significantly better at walking control, even producing recognizable gaits.

Algorithms↗

Determination of renal plasma flow by endogenous hippuric acid clearance in the conscious rat.

(1) The endogenous hippuric acid can be fluorometrically determined down to a minimum of 0.6 micrograms/ml in 10 microliters of rat serum. This enables the determination of the effective renal plasma flow (ERPF) by measurement of the renal clearance of the endogenous hippuric acid in the conscious rat. (2) Endogenous hippuric acid clearance (128 ml/min . m2 or 2.48 ml/min/100 g body weight) corresponds in its extent with the clearance after a single injection of unlabelled hippuric acid and is higher (20%) than the clearance after a single injection of 131I-hippuran. (3) Exogenous hippuric acid has a short biological half-time of 12-20 min and a distribution space of about 50%; therefore, alteration of its intermediary supply can disturb the estimate of the renal clearance of endogenous hippuric acid. (4) The daily rhythm of the renal excretion of endogenous hippuric acid shows a constant low excretion between 0800 and 1200 hours; this period is the best suited for clearance measurements.

Animals↗