Motion correction in MRI using an apparatus for dynamic angular position tracking (ADAPT).

Motion during MRI examinations is a serious problem that degrades the quality of the data (images) acquired. Motion can be corrected during the postprocessing of the data; however, this approach is suboptimal and is typically limited to in-plane or translational motion. An apparatus for dynamic angular position tracking (ADAPT) for prospective angular motion correction has been developed. This application is capable of "tracking" the scanned region of interest by performing dynamic adjustments of orientation of the scanning plane. The operation of the apparatus is based on deuterium MR spectroscopy and does not rely on the use of magnetic field gradients. Orientation-sensitive deuterium quadrupolar interaction in a single crystal attached to a subject is used to monitor the angular position in magnetic fields. Measurements are performed with an independent spectrometer channel in the background of the MRI scans. This apparatus is very cost- and time-efficient because it utilizes the hardware already available on many spectrometers and can be used in parallel with MRI scans. Potentially, rotations by a fraction of one degree can be easily corrected and the angular position information can be rapidly updated.

Dynamic publication model for neurophysiology databases.

We have implemented a pair of database projects, one serving cortical electrophysiology and the other invertebrate neurones and recordings. The design for each combines aspects of two proven schemes for information interchange. The journal article metaphor determined the type, scope, organization and quantity of data to comprise each submission. Sequence databases encouraged intuitive tools for data viewing, capture, and direct submission by authors. Neurophysiology required transcending these models with new datatypes. Time-series, histogram and bivariate datatypes, including illustration-like wrappers, were selected by their utility to the community of investigators. As interpretation of neurophysiological recordings depends on context supplied by metadata attributes, searches are via visual interfaces to sets of controlled-vocabulary metadata trees. Neurones, for example, can be specified by metadata describing functional and anatomical characteristics. Permanence is advanced by data model and data formats largely independent of contemporary technology or implementation, including Java and the XML standard. All user tools, including dynamic data viewers that serve as a virtual oscilloscope, are Java-based, free, multiplatform, and distributed by our application servers to any contemporary networked computer. Copyright is retained by submitters; viewer displays are dynamic and do not violate copyright of related journal figures. Panels of neurophysiologists view and test schemas and tools, enhancing community support.

High-throughput cytochrome P450 (CYP) inhibition screening via a cassette probe-dosing strategy. VI. Simultaneous evaluation of inhibition potential of drugs on human hepatic isozymes CYP2A6, 3A4, 2C9, 2D6 and 2E1.

The inhibition potential of drugs towards five major human hepatic cytochrome P450 (CYP) isozymes (CYP2A6, 3A4, 2C9, 2D6, and 2E1) was investigated via cassette dosing of the five probe substrates (coumarin, midazolam, tolbutamide, dextromethorphan, and chlorzoxazone) in human liver microsomes using a 96-well plate format. After microsomal incubations had been terminated with formic acid, the five marker metabolites (7-hydroxycoumarin, 1'-hydroxymidazolam, 4-hydroxytolbutamide, dextrorphan, and 6-hydroxychlorzoxazone) were simultaneously quantified using direct injection/online guard cartridge extraction/tandem mass spectrometry (DI-GCE/MS/MS). Several advantages resulted from the use of a short C(18) guard cartridge (4 mm in length) for DI-GCE/MS/MS, including minimal sample preparation, fast online extraction, short analysis time (2.5 min), and minimal source contamination. In addition, this method demonstrated an inter-day accuracy range from -8.7 - 7.4% with a precision less than 8.3% for the quantification of all the marker metabolites. The inhibition assay for the five CYP isozymes was evaluated using their known selective inhibitors via individual and cassette dosing of the probe substrates. The IC(50) values measured via cassette dosing were consistent with those observed via individual dosing, which were all in agreement with the reported values. In addition, the validated assay was used to evaluate the inhibitory potential of 23 generic drugs (randomly selected) towards the five CYP isozymes. The results suggest the integration of the cassette dosing strategy and the DI-GCE/MS/MS method can provide a reliable in vitro approach to screening the inhibitory potential of new chemical entities, with maximal throughput and cost-effectiveness, in support of drug discovery and development.

High-throughput cytochrome P450 (CYP) inhibition screening via cassette probe-dosing strategy. II. Validation of a direct injection/on-line guard cartridge extraction-tandem mass spectrometry method for CYP2D6 inhibition assessment.

A highly efficient direct injection/on-line guard cartridge extraction-tandem mass spectrometry (DI/GCE-MS-MS) method has been validated for high-throughput evaluation of cytochrome P450 (CYP) 2D6 inhibition potential using human hepatic microsomes and 96-well microtiter plates. Microsomal incubations were terminated with formic acid, centrifuged, and the resulting supernatants were injected for DI/GCE-MS-MS analysis. Due to the novel use of an extremely short C18 guard cartridge, this method exhibits several advantages, such as no sample preparation, excellent on-line extraction, short run time (2.5 min), and minimized source contamination and performance deterioration. The DI/GCE-MS-MS method demonstrates acceptable accuracy and precision for the quantification of dextrorphan, a marker metabolite of dextromethorphan mediated by CYP2D6, in microsomal incubations. The CYP2D6 inhibition assay has been validated using quinidine as a known selective inhibitor of the isoform. The IC50 value (0.20 microM) measured by the new method is in good agreement with the literature value (0.22 microM).

High-throughput cytochrome P450 (CYP) inhibition screening via cassette probe-dosing strategy: III. Validation of a direct injection/on-line guard cartridge extraction-tandem mass spectrometry method for CYP2C19 inhibition evaluation.

A new direct injection/on-line guard cartridge extraction-tandem mass spectrometry (DI/GCE-MS-MS) method has been validated for high-throughput evaluation of cytochrome P450 (CYP) 2C19 inhibition potential using human hepatic microsomes and 96-well microtiter plates. Microsomal incubations were quenched with formic acid, centrifuged, and the resulting supernatants were injected for DI/GCE-MS-MS analysis. Due to the use of a C(18) guard cartridge (4x2.0 mm I.D.), this method exhibits several advantages such as little sample preparation, rapid on-line extraction, short analysis time (2.5 min), and minimal source contamination and performance deterioration. The DI/GCE-MS-MS method demonstrates an inter-day accuracy ranged from 0.3 to 2.4% with precision ranging from 2.0 to 3.0% for the quantification of 4-hydroxymephenytoin, a marker metabolite of S-mephenytoin mediated by CYP2C19, in microsomal incubations. The CYP2C19 inhibition assay has been validated using tranylcypromine as a known inhibitor of the isoform. The IC(50) value (43.5 microM) measured by the new method is in agreement with a reported literature value (approximately 30 microM).

High-throughput cytochrome P450 (CYP) inhibition screening via a cassette probe-dosing strategy. V. Validation of a direct injection/on-line guard cartridge extraction--tandem mass spectrometry method for CYP1A2 inhibition assessment.

An efficient direct injection/on-line guard cartridge extraction-tandem mass spectrometry (DI/GCE--MS--MS) method has been validated for high-throughput evaluation of cytochrome P450 (CYP) 1A2 inhibition potential using human hepatic microsomes and 96-well microtiter plates. Microsomal incubations were terminated with formic acid, centrifuged, and the resulting supernatants were injected for DI/GCE--MS--MS analysis. Due to the use of an extremely short C(18) guard cartridge, this method offers several advantages such as no sample preparation, excellent on-line extraction, short run time and minimal source contamination and performance deterioration. The DI/GCE--MS--MS method demonstrates acceptable accuracy and precision for the quantification of resorufin, a marker metabolite of ethoxyresorufin mediated by CYP1A2, in microsomal incubations. The inhibition potential of CYP1A2 has been evaluated using its selective inhibitors, alpha-naphthoflavone and furafylline. The IC(50) values (120 nM for alpha-naphthoflavone and 5.1 microM for furafylline) measured by the new method are in agreement with the literature values.

Common data model for neuroscience data and data model exchange.

OBJECTIVE: Generalizing the data models underlying two prototype neurophysiology databases, the authors describe and propose the Common Data Model (CDM) as a framework for federating a broad spectrum of disparate neuroscience information resources. DESIGN: Each component of the CDM derives from one of five superclasses-data, site, method, model, and reference-or from relations defined between them. A hierarchic attribute-value scheme for metadata enables interoperability with variable tree depth to serve specific intra- or broad inter-domain queries. To mediate data exchange between disparate systems, the authors propose a set of XML-derived schema for describing not only data sets but data models. These include biophysical description markup language (BDML), which mediates interoperability between data resources by providing a meta-description for the CDM. RESULTS: The set of superclasses potentially spans data needs of contemporary neuroscience. Data elements abstracted from neurophysiology time series and histogram data represent data sets that differ in dimension and concordance. Site elements transcend neurons to describe subcellular compartments, circuits, regions, or slices; non-neuroanatomic sites include sequences to patients. Methods and models are highly domain-dependent. CONCLUSIONS: True federation of data resources requires explicit public description, in a metalanguage, of the contents, query methods, data formats, and data models of each data resource. Any data model that can be derived from the defined superclasses is potentially conformant and interoperability can be enabled by recognition of BDML-described compatibilities. Such metadescriptions can buffer technologic changes.

High-throughput cytochrome P450 inhibition screening via cassette probe-dosing strategy. IV. Validation of a direct injection on-line guard cartridge extraction/tandem mass spectrometry method for simultaneous CYP3A4, 2D6 and 2E1 inhibition assessment.

A highly efficient direct injection on-line guard cartridge extraction/tandem mass spectrometry (DI-GCE/MS/MS) method has been validated for high-throughput evaluation of cytochrome P450 (CYP) 3A4, 2D6 and 2E1 inhibition potential via cassette dosing of midazolam, dextromethorphan and chlorzoxazone using human hepatic microsomes and 96-well microtiter plates. Microsomal incubations were terminated with formic acid, centrifuged, and the resulting supernatants were injected for analysis by DI-GCE/MS/MS. Due to the novel use of an extremely short C(18) guard cartridge (4 mm in length), this method exhibits several advantages such as no sample preparation, excellent on-line extraction, short run time (2.5 min), and minimized source contamination and performance deterioration. The DI-GCE/MS/MS method demonstrates acceptable accuracy and precision for the simultaneous quantification of 1'-hydroxymidazolam, dextrorphan and 6-hydroxychlorzoxazone in microsomal incubations. The inhibition potential of CYP3A4, 2D6 and 2E1 has been evaluated using their known selective inhibitors. The IC(50) values measured by the cassette dosing approach (high-throughput) are consistent with those observed by an individual dosing regimen (conventional) and are all in good agreement with the literature values. The results suggest that the cassette probe-dosing strategy may provide an in vitro approach to minimize cost while maximizing throughput of CYP inhibition evaluation of new chemical entities in support of drug discovery and development.

High-throughput cytochrome P450 (CYP) inhibition screening via cassette probe-dosing strategy. I. Development of direct injection/on-line guard cartridge extraction/tandem mass spectrometry for the simultaneous detection of CYP probe substrates and their metabolites.

A highly efficient direct injection/on-line guard cartridge extraction/tandem mass spectrometry (DI-GCE/MS/MS) method utilizing electrospray polarity switching was developed for the simultaneous detection of probe substrates and marker metabolites of seven human hepatic cytochrome P450 (CYP) isozymes: CYP1A2, 2A6, 3A4, 2C9, 2C19, 2D6 and 2E1. Microsomal incubations were terminated with formic acid, centrifuged, and the resulting supernatants were injected for analysis by DI-GCE/MS/MS. This method employed an extremely short C(18) cartridge (4 mm in length) which allowed rapid cleanup of sample matrices while retaining the analytes an appropriate time (2. 0-2.2 min). From 1.5 to 2.7 min the effluent was directed to the mass spectrometer for detection otherwise diverted to waste. As a result of the efficient on-line extraction, matrix (e.g., salts and proteins) suppression was minimized. In addition, no visible source contamination was observed and system performance (chromatographic and mass spectrometric) did not significantly deteriorate after 500 consecutive injections. Electrospray polarity switching was strategically executed on a Micromass Quattro II mass spectrometer by establishing dummy ion transitions to protect the analytes from the interference of the overwhelming noise which was unavoidable for the first transition scanned following each polarity switch. This unique strategy led to the simultaneous detection of seven CYP probe substrates and seven corresponding marker metabolites (12 by positive mode and 2 by negative mode).

A novel method for storage of medical records and documents.

Many medical groups have a problem storaging and managing inactive medical records. The storage process involves selecting the records from inactive status, moving the records to storage and retrieving them if the patient is re-established. Many groups simply run out of storage space and are in need of an efficient and cost-effective way to manage this problem. This case study will show that an optical storage system provides a cost-effective method of archiving medical documents that can be easily indexed and retrieved.

Transport of thyrotropin releasing hormone in rabbit buccal mucosa in vitro.

The transport of thyrotropin releasing hormone (TRH) in rabbit buccal mucosa in vitro has been investigated with respect to (a) rate and type of metabolism of TRH on mucosal and serosal sides of buccal mucosa, (b) mechanism of TRH transport including charge effect on its permeability, and (c) pathway and rate-limiting regions of TRH movement. In addition, the integrity of excised buccal mucosa has been evaluated for purposes of in vitro solute diffusion experiments using tissue ATP level data, transmission electron microscopy, and TRH transport kinetic data. The results indicate that excised rabbit buccal mucosa can be used for TRH diffusion studies for approximately 6 hr. In addition, TRH apparently traverses buccal mucosa by simple diffusion with a steady-state permeability of about 10(-7) cm/sec, and this permeability is independent of pH. Moreover, the primary pathway appears to be via the intercellular space in the rate-limiting barrier, i.e., the upper 50 microns of the epithelium. Finally, TRH is degraded predominantly by deamidase activity, which is followed by, to a lesser degree, carboxypeptidase metabolism.

Pinworm infestation of the genital tract.

Extraintestinal enterobiasis has been reported in the vagina, endometrium, myometrium, ovary, fallopian tube and pelvic peritoneum. Gravid female pinworms may migrate from the perianal area to the vagina, ascend through the genital tract and exit through the fallopian tube to the peritoneum. Ectopic enterobiasis seldom causes clinical symptoms, but chronic granuloma formation may simulate other mass lesions in the pelvis.

An inexpensive servo-respirator based upon regulation of a shunt resistance.

We have designed and built an inexpensive servo-respirator for use in investigations of respiratory control in small animals. The device uses a butterfly valve to alter the resistance of an outflow shunt from a manifold that connects the animal's tracheal cannula to a pressure source. Tracheal pressure is regulated in response to a command provided by a suitably processed neural signal, often the integrated phrenic neurogram. As the valve opens, tracheal pressure approaches atmospheric; as it closes, tracheal pressure approaches the source pressure. An electronic controller circuit was developed to permit experimental procedures that include withholding volume delivery while maintaining a desired level of positive end-expiratory pressure. The device is able to track the neural command signal satisfactorily, and its performance appears to be limited primarily by the constraints applied by the respiratory system mechanics.

Retinoblastoma.

Retinoblastoma was diagnosed in 74 patients between July 1967 and February 1987. Thirty cases (40%) were bilateral; 39 were female and 35 were male. Treatment in bilateral cases consisted of enucleation of the more involved eye and a combined approach of supervoltage irradiation, cryocoagulation, and photocoagulation to the remaining eye. In unilateral cases the involved eye was enucleated and the uninvolved eye observed. Four patients had bilateral enucleation at their initial presentation and four others had the second eye enucleated after unsuccessful tumor treatment. One patient died from metastatic retinoblastoma, one patient died after intrathecal chemotherapy without the evidence of tumor, and one patient died with trilateral retinoblastoma. Seventy-one of 74 patients (96%) currently survive with a follow-up of 1 month to 19 years. Two-year survival is 65 of 68 (95.5%).

Differential depression of hypoglossal nerve activity by alcohol. Protection by pretreatment with medroxyprogesterone acetate.

Upper airway obstruction during sleep occurs more commonly in men than in women and has been treated with progestational agents with some success. Alcohol ingestion exacerbates sleep apnea, and recent studies have established that alcohol depresses the respiratory motor activity to upper airway muscles more than that to the diaphragm, a response pattern that favors upper airway obstruction during inspiration. To investigate the possibility that progesterone provides some protection from this action of alcohol, we studied the responses of phrenic and hypoglossal nerve activities to alcohol infusion in decerebrate cats pretreated with medroxyprogesterone acetate (MPA) or with control injections. The results indicate that pretreatment with MPA reduces the alcohol-induced mismatching of hypoglossal and phrenic activities. This action of MPA may contribute to its effectiveness in the treatment of some patients with inspiratory obstruction during sleep.

Inflammatory tumor response to monoclonal antibody infusion.

Two patients with melanoma and one with apudoma, all three with metastatic disease, received monoclonal antibody infusions with mAb R-24, specific for the disialoganglioside GD3. This marker was shown to be restricted to melanoma cells and a few other tumors of neural crest origin. Following treatment with mAb R-24 both melanoma patients showed inflammatory cutaneous responses around tumor nodules, i.e. blister formation or inflammatory perinodular halos. Local pain in bulky intestinal tumor sites occurred in all three patients about 3 hr after onset of antibody infusion. Adverse side-effects of antibody application were not observed with antibody doses up to 200 mg (single) and 440 mg total dose. The presented data indicate that mAb R-24 is active in vivo.

Strategies in the design of solution-stable, water-soluble prodrugs I: a physical-organic approach to pro-moiety selection for 21-esters of corticosteroids.

The ideal water-soluble prodrug should exhibit sufficient aqueous solution stability to allow long-term storage of its solutions (i.e., 2 years at room temperature) and yet should be converted rapidly in vivo to the active parent drug--two severe and seemingly conflicting demands which limit the utility of many common solubilizing pro-moieties. For example, succinate esters, which are commonly utilized as water-soluble prodrugs, are unstable in solution and may undergo slow and incomplete bioconversion in vivo. In this study, the solution stability problems associated with 21-esters of corticosteroids are reviewed. It is concluded that the most important reaction limiting shelf life is ester hydrolysis. From a consideration of the influence of molecular structure on ester reactivity, a strategy for the design of solution-stable, water-soluble prodrugs of corticosteroids has been developed. Two key requirements for dilute solution stability are high solubility at the pH of optimum stability and appropriate design of the pH-rate profile. Several 21-esters of methylprednisolone have been synthesized, and the rates of their aqueous solution hydrolysis have been determined to test the strategy. Compounds exhibiting estimated shelf lives in dilute solution of greater than 2 years at 25 degrees C have been identified.

Strategies in the design of solution-stable, water-soluble prodrugs II: properties of micellar prodrugs of methylprednisolone.

In a previous study, a physical-organic approach to the design of solution-stable, water-soluble prodrugs of the corticosteroid methylprednisolone was outlined, and several 21-esters were synthesized to test the approach. Compounds exhibiting dilute solution stabilities approaching 2 years at 25 degrees C were reported. A complicating factor in more concentrated aqueous solutions of water-soluble prodrugs, however, is the limited extent to which hydrolysis can occur before the solution becomes saturated with respect to the relatively insoluble parent drug. In this study the advantages of micellar prodrugs as water-soluble delivery systems for parenteral administration of relatively insoluble parent drugs are explored. Micellar prodrugs, besides being highly water soluble, have additional advantages in that their micelles solubilize poorly soluble degradation products which may otherwise precipitate and may act as a self-stabilizing influence due to protection of the hydrolytically labile prodrug linkage within the micelle interior. Two 21-esters of methylprednisolone previously identified as having promising dilute solution stability have now been shown to self-associate in aqueous solution at higher concentrations, as determined by solubility, kinetic, and light-scattering measurements. One consequence of self-association is that free methylprednisolone, the product of prodrug hydrolysis, is solubilized in concentrated prodrug formulations. In addition, acid- and base-catalyzed hydrolysis rate constants are altered in the micelles, resulting in further prolongation of shelf life in concentrated solutions. Due to the added benefits of self-micellization, the water-soluble 21-esters investigated exhibit shelf lives exceeding 2 years at 30 degrees C, the upper limit of the controlled room temperature range.

Dynamic changes of hypoglossal and phrenic activities by hypoxia and hypercapnia.

Steady-state responses of unanesthetized subjects to hypercapnia or hypoxia provide no evidence that chemoreceptor afferents are differentially distributed upon hypoglossal and bulbospinalphrenic neurons, as suggested by results in anesthetized animals. We hypothesized that dynamic changes in activities of phrenic and hypoglossal nerves might differ following sudden alterations of inspired gases. This hypothesis was based on the observation that episodes of obstructive apnea may follow central apnea. The obstruction might reflect phrenic activity increasing more quickly than that of the hypoglossal upon resumption of ventilation. In decerebrate, vagotomized, paralyzed and ventilated cats, we recorded phrenic and hypoglossal activities before and during abrupt, sustained exposure to hypercapnia, hypoxia and hypoxic hypercapnia, and following sudden withdrawal of these stimuli. For all such manoeuvres, activities of phrenic and hypoglossal nerves increased or decreased in parallel fashion. Our findings cause rejection of the hypothesis of differing dynamic phrenic and hypoglossal responses to chemoreceptor stimuli. The concept that the ventilatory control system is well organized to prevent upper airway obstructions is discussed.