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Quantitative ultrastructural immunocytochemistry using a computerized image analysis system.

Secretory granules in human pituitary adenoma cells have been examined indirectly for hormone epitopes by immunogold labelling of resin-embedded ultrathin sections. The specific binding of different immunoglobulin-gold complexes to the antigrowth hormone antibodies over the secretory granules was measured using a computerized image analysis system. This facilitated the assessment of the preferential binding to the target granules of gold particles with three different average particle diameters (Au7, Au11, Au17). The time of pretreatment of sections with H2O2 or a buffer was found to influence the staining considerably. The scanning electron microscopic findings of protruded secretory granules with a mountain-like surface might be relevant to the uneven distribution of immunolabels seen over the secretory granules in the adenohypophysis.

Adenoma↗

[Quantifying the damage to conjunctival and corneal cell cultures caused by uv light using CASY (Cell Analysis System). A method for reducing animal experiments].

BACKGROUND: By depletion of stratospheric ozone, enhanced levels of UV radiation reach the surface of the earth. Exposure of the anterior parts of the eye to UV radiation leads to irritation of the conjunctival and corneal cells. METHOD: By the CASY (cell analysis) system the influence of UV radiation on cultures of conjunctival and corneal cells was observed by determination of the cell counts, cell diameter, and the cell volume. RESULTS: By comparing these parameters with the control, damage of the conjunctival and corneal cells by UV radiation can be determined within 2 s of exposure of the cells in quartz glass vials to the UV light. CONCLUSION: By the CASY system the dramatic influence of UV radiation on cells of the anterior parts of the eye can be determined. This system enables objective statements on the cytotoxicity of radiation, chemical substances and drugs on cell cultures without the use of radioactive methods, complicated determination of cell metabolism or staining methods which are difficult to standardize. Also, studies on animals can be reduced by the CASY system.

Animal Testing Alternatives↗

A systems analysis of importin-{alpha}-{beta} mediated nuclear protein import.

Importin-beta (Impbeta) is a major transport receptor for Ran-dependent import of nuclear cargo. Impbeta can bind cargo directly or through an adaptor such as Importin-alpha (Impalpha). Factors involved in nuclear transport have been well studied, but systems analysis can offer further insight into regulatory mechanisms. We used computer simulation and real-time assays in intact cells to examine Impalpha-beta-mediated import. The model reflects experimentally determined rates for cargo import and correctly predicts that import is limited principally by Impalpha and Ran, but is also sensitive to NTF2. The model predicts that CAS is not limiting for the initial rate of cargo import and, surprisingly, that increased concentrations of Impbeta and the exchange factor, RCC1, actually inhibit rather than stimulate import. These unexpected predictions were all validated experimentally. The model revealed that inhibition by RCC1 is caused by sequestration of nuclear Ran. Inhibition by Impbeta results from depletion nuclear RanGTP, and, in support of this mechanism, expression of mRFP-Ran reversed the inhibition.

Active Transport, Cell Nucleus↗

Development of an improved automated gas-chromatographic chiral analysis system: application to non-natural amino acids and natural protein hydrolysates.

In the use of combinatorial chemistry as a novel strategy for drug discovery, the chirality assessment of building blocks used for library construction is particularly important in the evaluation of biological actions of generated libraries. The procedure for chiral analysis of lead compounds in screening may be of a high priority, particularly in the case of the protection of intellectual rights. Previously, an automated amino acid analysis system using enantiomer labeling was developed. The system incorporates a reactor, which allows automated esterification and acylation of amino acids, and is connected to an on-line gas chromatographic system. A capillary column coated with a chiral phase is employed for the separation of the enantiomers. This particular system is improved with a newly constructed "high-throughput auto-derivatizer" in combination with a new GC-system. The resulting data can be processed by newly constructed software. The analyses of amino acid derivatives or hydrolysates of proteins and peptides are carried out routinely within ca. 45 min, including derivatization. Using this system several non-natural amino acids were tested with respect to the stereoisomeric configuration. In addition, acid hydrolysates of food proteins and tissues obtained by autopsy were analyzed as an application to proteome research.

Aged↗

A computer image analysis system for microvessel density measurement in solid tumours.

Microvessel density (MVD) is a widely used surrogate measure of angiogenesis in pathological specimens and tumour models. Measurement of MVD can be achieved by several methods. Automation of counting methods aims to increase the speed, reliability and reproducibility of these techniques. The image analysis system described here enables MVD measurement to be carried out with minimal expense in any reasonably equipped pathology department or laboratory. It is demonstrated that the system translates easily between tumour types which are suitably stained with minimal calibration. The aim of this paper is to offer this technique to a wider field of researchers in angiogenesis.

Blood Vessels↗

Cognitive-behavioral analysis system of psychotherapy as a maintenance treatment for chronic depression.

Although the efficacy of maintenance pharmacotherapy for the prevention of recurrence in major depressive disorder (MDD) is well documented, few studies have tested the efficacy of psychotherapy as a maintenance treatment. The authors examined the efficacy of the cognitive-behavioral analysis system of psychotherapy (CBASP) as a maintenance treatment for chronic forms of MDD. Eighty-two patients who had responded to acute and continuation phase CBASP were randomized to monthly CBASP or assessment only for 1 year. Significantly fewer patients in the CBASP than assessment only condition experienced a recurrence. The 2 conditions also differed significantly on change in depressive symptoms over time. These findings support the use of CBASP as a maintenance treatment for chronic forms of MDD.

Adolescent↗

[The study of automatic cephalometric analysis system].

OBJECTIVE: To develop a system of automatic cephalometric analysis and compare the reliability of the computer software systems (ACAS and OSPES). METHODS: The establishment of the ACAS computer software system by image processing technique was introduced. 15 cephalograms were used for comparing the reliability of cephalometry between ACAS and OSPES. RESULTS: The results shown that the ACAS is more reliable for soft tissue profile measurement than OSPES. CONCLUSION: The application of ACAS would supply a valuable measurement tool for orthodontics and orthognathic surgery.

Cephalometry↗

A rapid method of screening for N-acetyltransferase (NAT2) phenotype by use of the WAVE DNA fragment analysis system.

N-Acetyltransferase 2 (NAT2) is involved in Phase II biotransformation of a variety of toxicants. Polymorphisms in the NAT2 gene result in a slow acetylator phenotype, which has been associated with various cancers and neurodegenerative diseases. To date most studies investigating NAT2 genotype/phenotype have adopted an RFLP approach, which is both expensive and time-consuming. Using the Wave DNA fragment analysis system, we have developed a fast and robust method of identifying two polymorphisms (C282T and T341C) of the NAT2 gene which allows identification of the most common slow acetylator alleles found in Caucasian populations: NAT2*5, NAT2*6, NAT2*7, and NAT2*14. This was done by comparing phenotype status in 126 samples genotyped by RFLP analysis and also by Wave analysis for the polymorphisms C282Tand T341C. All 126 samples analyzed by both RFLP and Wave analysis gave consistent phenotype results and 100% correlation was achieved between the two methods.

Arylamine N-Acetyltransferase↗

[Use of a computer-controlled kinematic-electromyographic motion analysis system to investigate gait patterns of healthy subjects on the level walkway and the treadmill (author's transl)].

In an attempt to clarify the question if the level walkway and the treadmill present comparable conditions for analytic examinations of the human gait we investigated in either situation with a computer-controlled motion analysis system some kinematic and electromyographic aspects of the gait pattern of 20 healthy subjects. The results show that on the treadmill certain kinematic events in the time pattern of the gait cycle are shifted towards the beginning of the full stride. Moreover, in the treadmill walk the amplitude of the angular motion of the thigh is diminished in the forward and enhanced in the backward direction, while the angular amplitude of the shank is diminished in either direction. As to the electromyographic results, we found in the treadmill walk a prolonged activity of the gluteus maximus, and adductor magnus, the semitendinosus, and the gastrocnemius muscle. These results show that there may exist significant kinematic and electromyographic differences between the treadmill and the normal level walk. This fact has to be considered whenever a treadmill is considered for gait analytical investigations.

Adult↗

Study of imidaclopride removal from aqueous solution by adsorption onto granular activated carbon using an on-line spectrophotometric analysis system.

The removal of imidaclopride as a pesticide by granular activated carbon (GAC) and its adsorption kinetics were studied at different pH values and temperatures. In all experiments, the amount of GAC and initial concentration of imidaclopride were 2g and 25 ppm, respectively. The adsorption process was followed by an on-line spectrophotometric analysis system, which consisted of UV-spectrophotometer, a designed absorption cell, peristaltic pump and special glassy reactor. The effect of pH and temperature on adsorption was studied over 90 min adsorption periods. The obtained data were treated according to various kinetic models. The results showed that second order model was the most suitable one on the overall. The our results also showed that the adsorption rate constants for first order, second order and intraparticle diffusion models followed decreasing order: pH=7>4>10>1, T=25>35>45>55 degrees C.

Adsorption↗

Electrochemical determination of gamma-glutamyl transpeptidase activity and its application to a miniaturized analysis system.

A novel method to determine the activity of gamma-glutamyl transpeptidase (gamma-GTP) was developed. gamma-l-glutamyl-l-glutamate and glycyl-glycine were used as the substrates for gamma-GTP. l-glutamate produced by the enzymatic reaction was measured with an amperometric l-glutamate sensor. Following the mixing of the substrate solution and a sample solution, the current generated on the l-glutamate sensor continued to increase at a constant rate. The method was used to construct a miniaturized analysis system for the determination of gamma-GTP activity. The system consisted of the l-glutamate sensor formed on a glass substrate and a polydimethylsiloxane (PDMS) flow channel. Since the l-glutamate concentration in the solution increased as the solution was mobilized through the flow channel, a constant current increase was observed. The relation between the slope of the response curve and the activity of gamma-GTP was linear between 35 U l(-1) and 659 U l(-1). The rate analysis in the micro flow channel minimized the influence of interferents. The reproducibility of the output of the micro system was found to be good with a relative standard deviation (R.S.D.) of 5.6% at 659 U l(-1). The activities of gamma-GTP in human serum samples were also determined and compared with values obtained with a conventional spectroscopic method. The values obtained by the two methods were consistent with a correlation coefficient of 0.953.

Biomarkers↗

Symptomatic and syndromal anxiety in chronic forms of major depression: effect of nefazodone, cognitive behavioral analysis system of psychotherapy, and their combination.

BACKGROUND: Limited information is available on treatment response of anxiety symptoms in chronic forms of major depression. Concurrent anxiety disorders are prevalent in chronic depression, but the responsiveness of patients with such comorbidity to different treatments is largely unknown. This study investigated the comparative efficacy of nefazodone, Cognitive Behavioral Analysis System of Psychotherapy (CBASP), and their combination in improving anxiety symptoms in patients with chronic forms of major depression, including those with a concurrent anxiety disorder. METHOD: 681 patients with chronic major depressive disorder (DSM-IV criteria) participated in a multicenter study of 12 weeks of acute treatment with nefazodone (N = 226), CBASP (N = 228), or the combination (N = 227). The Hamilton Rating Scale for Anxiety (HAM-A), the HAM-A psychic anxiety factor, and the anxiety/arousal subscale of the 30-item Inventory for Depressive Symptomatology-Self Report (IDS-SR-30) were used to assess anxiety symptoms. RESULTS: In the full sample. without controlling for change in depressive symptoms, combination therapy was superior to both monotherapies on all 3 anxiety measures both in the rate of change and at endpoint. When change in depressive symptoms was controlled for, there were no treatment differences in rate of change from baseline to week 12 on any of the 3 anxiety measures. In those patients with a concurrent anxiety disorder, however, the combination was superior to CBASP on the HAM-A and the IDS-SR-30. Nefazodone alone and combination therapy were both superior to CBASP on the HAM-A psychic anxiety factor. CONCLUSION: For patients with chronic depression, combination therapy is superior to CBASP or nefazodone alone. Among patients with a concurrent anxiety disorder, nefazodone. either alone or in combination with CBASP, improves anxiety symptoms faster than CBASP alone, independent of depressive symptom reduction.

Adult↗

Quantitative computation of induced heterotopic bone formation by an image analysis system.

The formation of heterotopic bone (HO) is an unsolved problem and occurs in response to a variety of conditions including muscle contusions, injuries, surgical procedures, and burns. A sensitive method for quantification of HO would be valuable for experimental work and research on bone morphogenetic protein (BMP). The hardware and software used in an image analysis system are presented. This system provides a rapid and highly accurate method for determining the amount of HO in a muscle. Using the mouse thigh pouch as an experimental system, this method demonstrated that quantities of new bone were formed in direct proportion to the amount of BMP/NCP implanted. This system can be used for measuring HO on roentgenograms or any other image, including computed tomographic (CT) scans, magnetic resonance imaging (MRI), and autoradiograms.

Animals↗

Flow injection analysis system for the supervision of industrial chromatographic downstream processing in biotechnology.

Sugar beet molasses is a natural resource for various products used in daily life, ranging from sucrose to amino acids for pharmaceutical industry. The separation of molasses into these high value components is performed on a large scale by ion exchange/exclusion chromatography. A biosensor system was set up for the "in time" analysis of serine and sucrose during molasses desugarisation. D-Serine was analysed with the multi-enzyme system D-serine dehydratase/lactic dehydrogenase and photometric detection of the NADH consumed. Sucrose was determined with invertase/mutarotase/glucose oxidase and the oxygen consumed was monitored amperometrically. An analysis could be performed within 2-5 min by directly injecting samples from the chromatographic process into the flow injection analysis system. The determination range for the sucrose analysis was 0-2.5 gl-1 and for the analysis of D-serine 0-0.5 gl-1. The standard deviation for the measurement of D-serine was 1.7%.

Biosensing Techniques↗

A distributed medical record on a Holter-ECG archiving and analysis system.

At the Department of Cardiology of the Technical University of Munich and the Deutsches Herzzentrum München cardiac disease patients are treated in close co-operation. In order to support the collaborative treatment as well as cardiological studies a distributed medical record (DMR) has been implemented on the basis of a Holter-ECG archiving and analysis system. Architecture, experiences and major benefits of the solution are reported in the paper. Between January 1995 and December 1997 6500 Holter-ECGs have been archived and analysed. The DMR is in routine operation since January 1996 for all heart catheter patients.

Attitude to Computers↗