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Biomedical subjects

C R Clark

Publications and source records attributed to C R Clark.

At least 55 records · Page 3Linked to original sources

Amplified enzyme-linked-immunofilter assays enable detection of 50-10(5) bacterial cells within 1 hour.

Two enhanced enzyme-linked-immunofilter assay (ELIFA) methods for the rapid and quantitative detection of whole bacterial cells are described. In the first method, specific antibody bound to bacterial cells was amplified using a secondary antibody and detected by the conjugated enzyme activity (peroxidase) of a third antibody in a chemiluminescent assay. In the second method, a chromogenic substrate was used in conjunction with a biotinylated secondary antibody and avidin. Both assays were conducted within 55 min using a 96-well continuous flow immunofilter apparatus. The assay values were determined either as the reflectance of developed X-ray film placed over chemiluminescent membranes or of precipitated chromogen on the membrane surface. The biotin/avidin method enabled quantitative detection of approximately 60 to 10(5) cells. The detection limit (blank + 2 SD) of the chemiluminescent assay with a 30-s film exposure time was 50 cells. The ELIFA methods described represent a considerable advance in sensitivity over previous immunological methods of detecting whole bacterial cells and suggest that immunological methods may approach PCR in sensitivity.

Antibodies, Bacterial↗

Creating information messages for reducing patient distress during health care procedures.

Health care providers want strategies which they can use to assist patients in decreasing their distress during threatening health care procedures. Concrete objective information has been shown in laboratory and clinical studies to be effective in reducing negative emotional responses. Concrete objective information is a message, given before a health care event, which describes the experience from the patients' point of view in unambiguous, concrete, and objective terms. According to self-regulation theory, this type of information facilitates coping by decreasing the difference between expectations and actual experience and by increasing patients' understanding of their experience. However, little emphasis has been placed in the literature on the process of creating the message. This paper will describe a four-stage process that permits systematic development of comprehensive and accurate concrete objective information messages to prepare patients for health care procedures.

Communication↗

Development of a gas chromatographic-mass spectrometric drug screening method for the N-dealkylated metabolites of fentanyl, sufentanil, and alfentanil.

A sensitive, specific urinary assay for fentanyl, sufentanil, and alfentanil based on their N-dealkylated metabolites is described. Norfentanyl, norsufentanil-noralfentanil, and 2H5-norfentanyl are synthesized and characterized by standard analytical techniques. Derivatization of these secondary amines to yield the pentafluorobenzamides produces stable products with good gas chromatographic properties and unique, high-mass fragments in their mass spectra. These properties are utilized to develop a drug screening procedure based on gas chromatography-mass spectrometry to detect these major metabolites in human urine. The metabolites are isolated from urine samples by a liquid-liquid extraction procedure. The method allows for detection of metabolite concentrations as low as 0.3 ng/mL.

Alfentanil↗

A rapid and sensitive enzyme linked immunofilter assay (ELIFA) for whole bacterial cells.

An improved method is described for the detection of Escherichia coli by an enzyme linked immunofilter assay (ELIFA) using nitrocellulose membrane sandwiched between two 96-well plates. The incorporation of a pumping system permits a continuous flow of reagents and/or wash fluids through the membrane and provides an assay procedure capable of detecting 10(3) bacteria per well within 40 min. Quantitative bacterial detection was based on precipitated chromogen determined by scanning densitometry. The procedure represents a significant improvement in assay time and/or sensitivity over previously described ELIFA and ELISA methods for whole bacterial cells.

Escherichia coli↗

Enlarged frontal P300 to stimulus change in panic disorder.

This study investigated event-related potential (ERP) indices of information processing in sufferers of panic disorder (PD). ERPs were recorded from 14 PD patients and 15 controls during an auditory target detection task. The task required subjects to discriminate infrequent target tones (p = .14; 2000 Hz) from frequent (p = .72; 1000 Hz) and infrequent (p = .14; 500 Hz) distractor tones. A frontal P300 (P3a) identified in the PD group was characteristic of activity that would be expected to novel, task-irrelevant stimuli and is consistent with junctional pathology involving the prefrontal-limbic pathways. This study provides psychophysiological evidence of an abnormality in PD of the brain's processing of physical changes in the stimulus field that occurs even under conditions of low stimulus load. It may assist in helping to understand the breakdown in information processing that occurs in PD under high load conditions such as crowds and supermarkets.

Adult↗

Profilin forms tetramers that bind to G-actin.

Profilin binds to G-actin and affects polymerization. However, regulation of profilin function is generally unknown and controversy exists regarding profilin effects on actin polymerization. Because protein-protein interactions are implicated in many cellular responses, human platelet profilin self-association and actin inter-action was examined. Silver stained SDS-PAGE of poly-l-proline/sepharose 4B column purified profilin revealed the presence of profilin (14.8 kD) and extraneous higher bands (primarily 30 kD and 58.5 kD). Re-electrophoretic analysis of gel electroelution purified profilin yielded predominantly 14.8 kD and 58.5 kD proteins. Rabbit IgG antibodies made against gel electroelution-purified profilin recognized all profilin sizes on immunoblots. Capillary electrophoresis of profilin in solution produced a single peak that resolved into three distinct peaks upon addition of reducing agent or high salt conditions. Further, G-actin did not bind to 14.8 kD profilin on immunoblot overlay assays, but surprisingly bound only to 58.5 kD profilin. The data indicate that monomeric profilin forms tetramers which are the relevant high affinity actin-binding form.

Actins↗

Notational analysis on game strategy used by the world's top male squash players in international competition.

The purpose of this study was to provide a profile of the competition strategy used by the world's top squash players at the international level and provide recommendations for other players to improve their playing strategy. A total of 10 matches including three in round one, the four quarter finals, the two semi-finals, and the final of the 1993 Hong Kong Squash Open were filmed using a 3-CDD video camera. The tournament was played under the North American scoring system. Notational analysis which is based on frame by frame video analysis was used to categorize a player's motion. The frequency of each stroke, and the success or failure rate of each stroke were recorded. Shots were classified as "effective", "ineffective", "winning" and "losing" shots and the analysis demonstrated that 70.28% were "effective", 19.86% were "ineffective", 5.37% were "winning" and 4.48% were "losing" shots. The mean number of shots per game was 252.09. The order of priority using different kinds of strokes in matches were as follows: drive (length) (60.79%), drop (17.91%), volley (11.79%), boast (4.80%), and lob (4.72%). The results of this study show that the "pressure and attack game" was the most important strategy for the world's top squash players in producing winning performances. Also identified are the strokes and the strategies which should be practiced to improve performance. This method of notational analysis can be utilized to evaluate the strengths and weaknesses of players at all levels of competition.

Competitive Behavior↗

Artificial neural networks: a prospective tool for the analysis of psychiatric disorders.

Artificial neural networks are computer simulations of biological parallel distributed processing systems. They are able to undertake complex pattern recognition tasks, including diagnostic classification, prediction of disease onset and prognosis, and identification of determinants of clinical decisions. These capabilities have been utilized in general medicine, but as yet there has been little application of artificial neural networks in psychiatric research. Artificial neural networks can also be used to create models of brain function, providing a paradigm for cognition and the organization of neural systems that demonstrates how changes at the cellular level can affect information processing. These models are able to encompass both the biological and the behavioral dimensions of psychiatric disorders.

Humans↗

An immunofiltration apparatus for accelerating the visualization of antigen on membrane supports.

Although proteins can be transferred and bound to a membrane with several different methods such as capillary transfer, electroblotting, etc., only the "shaker/incubation" method is commonly used for visualization of the proteins. We have tested an apparatus for the immunofiltration of solutions through nitrocellulose membrane which greatly accelerates the kinetics for the visualization of proteins. As a model system, avidin, bound on nitrocellulose membrane, was detected with 5-min filtering steps of a 1:2000 dilution of ascites of murine monoclonal antibody against avidin followed by a 1:5000 dilution of goat anti-mouse IgG-horseradish peroxidase and 0.5 mg/ml chloronapthol in the presence of 0.01% peroxide. The same solutions used with 5-min incubation steps with the shaker/incubation method could not detect avidin at almost 10 times that amount. Further studies with monoclonal antibodies specific for native C-reactive protein and modified-CRP, 15.1D6 and 13.3H12 respectively, showed that immunofiltration did not result in altered specificity compared to the shaker/incubation method. Also, data are presented showing the advantages of a 10-slot top for the immunofiltration of solutions through distinct areas of a membrane.

Antibodies, Monoclonal↗

Physiological profiles and sport specific fitness of Asian elite squash players.

There is a scarcity of descriptive data on the physiological characteristics of elite Asian squash players. The purpose of this study was to evaluate the physiological profile and sports specific fitness of Hong Kong elite squash players. It was conducted before the selection of the Hong Kong national squash team for the 1992 Asian Squash Championship. Ten elite squash players were selected as subjects for the study. Maximum oxygen uptake was measured using a continuous treadmill running test. A sports specific field test was performed in a squash court. The following means (s.d.) were observed: height 172.6(4.3) cm; weight 67.7(6.9) kg; body fat 7.4(3.4)%; forced vital capacity (FVC) 5.13(0.26) litres; maximum oxygen uptake (VO2max) 61.7(3.4) ml.kg-1.min-1; anaerobic threshold (AT) 80.2(3.3)% of VO2max; alactic power index 15.5(1.8) W.kg-1; lactic work index 323.5(29.4) J.kg-1, peak isokinetic dominant knee extensor and flexor strengths 3.11(0.29) Nm.kg-1 and 1.87(0.18) Nm.kg-1. The results show that the Hong Kong squash players have relatively high cardiorespiratory sports specific fitness and muscle strength which may be one of the key factors that contributed to the success of the Hong Kong team in the Asian Championship.

Adipose Tissue↗

Rheumatoid involvement of the cervical spine. An overview.

OBJECTIVE: This symposium is an update of current knowledge regarding rheumatoid involvement of the cervical spine. SUMMARY OF BACKGROUND DATA: The pathophysiology of rheumatoid involvement in the neck is the same as it is in the peripheral joints. Information regarding the natural history of this condition is incomplete, and more data are needed regarding the natural history in the untreated patient as well as the outcomes of operative management. CONCLUSIONS: Long-term studies of the natural history in the untreated patient as well as the results of operative management will be needed before definitive statements can be made regarding treatment recommendations. However, the latest information available is being presented to guide treatment at this time.

Arthritis, Rheumatoid↗

Spinal motion after cervical fusion. In vivo assessment with roentgen stereophotogrammetry.

STUDY DESIGN: A roentgen stereophotogrammetric (RS) technique using metallic markers was used to determine in vivo three-dimensional (3-D) motion data in the post-operative cervical spine. Two patients were examined with RS following fusion for up to 12 months at scheduled intervals. OBJECTIVES: The study was designed to develop a technique and provide an in vivo assessment of the fusion process following surgery. Besides the researchers and surgeons, this type of information is of particular interest to the FDA and implant manufacturers. SUMMARY OF BACKGROUND DATA: The high accuracy of the roentgen stereophotogrammetric (RS) technique using metallic markers to predict displacements across body joints, including spinal joints, has been well established in the literature. Its applications in the lumbar region to assess the fusion process as a function of time following surgery have also been reported. Similar in vivo applications (and results) dealing with the cervical region are lacking. METHODS: A roentgen stereophotogrammetric (RS) technique using metallic markers was developed. Appropriate in vitro studies were undertaken to assess its accuracy for in vivo applications in the cervical spine region. Following this, three Vitallium beads were surgically implanted in anatomically appropriate positions in of each the exposed vertebrae of two patients at the time of surgery. The patients were followed with RS serially for 12 months after surgery at scheduled intervals. At each RS session, roentgen stereo pairs of the cervical spine in neutral, maximum voluntary flexion, and maximum voluntary extension were obtained using a biplanar radiographic system. The metallic beads on the radiographs were digitized using an in-house software package to determine vertebral motions across the fused segments. RESULTS: In patient 1 (atlantoaxial fusion), the flexion-extension range of rotational motion decreased with time while the corresponding AP translation at the fusion level increased from 6 mm at 3 months post-op to 13 mm at 6 and 12 months postoperatively. The abnormal AP translation at 6 months and beyond was evident on extension/extension lateral radiographs, but the AP translation at 3 months was not visually evident. In patient 2 (anterior discectomy and interbody fusion), motions of small magnitude were observed at the fused level in all three axes. These motions actually increased over the one year observation period, but were never large enough to be visually detectable on lateral extension/extension radiographs. Patient 2 had a good clinical result despite these small motions. CONCLUSIONS: Roentgen stereophotogrammetry may detect motion in the in vivo cervical spine with a sensitivity heretofore unavailable. In patient 1, we were able to detect motion before it was visually manifest. In patient 2, small motions were detected at the level of a successful anterior cervical fusion, the significance of which remain uncertain.

Atlanto-Axial Joint↗

The pharmacokinetics and liver metabolism of N-hydroxy-3,4-methylenedioxyamphetamine (N-OH MDA) in rats.

The metabolism and disposition of N-hydroxy-3,4-methylenedioxyamphetamine (N-OH MDA) was studied by utilizing rat liver slices as well as by intravenous pharmacokinetic studies in rats. In the liver slice experiments, N-OH MDA (16 micrograms/ml) was incubated with rat liver slices and the disappearance of N-OH MDA and the appearance of MDA were observed over 2 hours. Drug and metabolites were assayed by a reverse phase high performance liquid chromatographic procedure including a C8 column and UV detection. N-OH MDA disappeared from the plasma at an apparent first-order rate with a t1/2 of 36.3 +/- 8.4 min. By the end of 2 hours, approximately 85% of the N-OH MDA was biotransformed to MDA. Following placement of jugular cannulas in rats, a dose of 15 mg/kg of N-OH MDA was administered intravenously. With rapid blood sampling, a t1/2 of 0.86 +/- 0.12 min was observed for N-OH MDA. A total body clearance of 9.09 +/- 2.06 L/hr/kg and a volume of distribution of 0.184 +/- 0.022 L/kg were noted. Plasma concentrations of MDA were observed after the N-OH MDA dose to rats. The MDA t1/2 after N-OH MDA dosing was 1.68 +/- 0.29 hours with peak plasma concentrations at 8 min. No other major metabolites could be detected in liver slice, plasma, or urine samples. Both sets of studies demonstrate the rapid conversion of N-OH MDA to MDA in rats.

3,4-Methylenedioxyamphetamine↗