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Phage lambda cDNA cloning vectors for subtractive hybridization, fusion-protein synthesis and Cre-loxP automatic plasmid subcloning.

We describe the construction and use of two classes of cDNA cloning vectors. The first class comprises the lambda EXLX(+) and lambda EXLX(-) vectors that can be used for the expression in Escherichia coli of proteins encoded by cDNA inserts. This is achieved by the fusion of cDNA open reading frames to the T7 gene 10 promoter and protein-coding sequences. The second class, the lambda SHLX vectors, allows the generation of large amounts of single-stranded DNA or synthetic cRNA that can be used in subtractive hybridization procedures. Both classes of vectors are designed to allow directional cDNA cloning with non-enzymatic protection of internal restriction sites. In addition, they are designed to facilitate conversion from phage lambda to plasmid clones using a genetic method based on the bacteriophage P1 site-specific recombination system; we refer to this as automatic Cre-loxP plasmid subcloning. The phage lambda arms, lambda LOX, used in the construction of these vectors have unique restriction sites positioned between the two loxP sites. Insertion of a specialized plasmid between these sites will convert it into a phage lambda cDNA cloning vector with automatic plasmid subcloning capability.

Bacteriophage lambda↗

The automatic implantable cardioverter-defibrillator: efficacy, complications and survival in patients with malignant ventricular arrhythmias.

Ninety-four patients underwent surgery for automatic implantable cardioverter-defibrillator implantation. Ninety patients were discharged from the hospital with the device and were followed up for a mean period of 17 +/- 10 months. Forty-six patients experienced at least one discharge of the device under circumstances consistent with a malignant ventricular arrhythmia. One sudden death occurred. Complications included perioperative death (3 patients), post-operative ventricular tachycardia (12 patients) and atrial fibrillation (8 patients), perioperative myocardial infarction (1 patient) and device discharges for sinus tachycardia and supraventricular arrhythmias (17 patients). Six and 12 month survival rates by life table analysis were 98.7 and 95.4%, respectively. Thus, the automatic implantable cardioverter-defibrillator is a highly effective and relatively low risk treatment modality for patients with refractory life-threatening ventricular arrhythmias.

Adolescent↗

Long-term outcome with the automatic implantable cardioverter-defibrillator.

The automatic implantable cardioverter-defibrillator was implanted in 270 patients because of life-threatening arrhythmias over a 7 year period. There was a history of sustained ventricular tachycardia or fibrillation, or both, in 96% of these patients, 80% had one or more prior cardiac arrests and 78% had coronary artery disease as their underlying diagnosis. The average ejection fraction was 34%, and 96% of these patients had had an average of 3.4 antiarrhythmic drug failures per patient before defibrillator implantation. There were four perioperative deaths and eight patients had generator infection or generator erosion, or both, during the perioperative period or during long-term follow-up. Concomitant antiarrhythmic drug therapy was given to 69% of patients. Shocks from the device were given to 58% of patients. and 20% received "problematic" shocks. The device was removed from 16 patients during long-term follow-up for a variety of reasons. There were 7 sudden cardiac deaths and 30 nonsudden cardiac deaths, 18 of which were secondary to congestive heart failure. The actuarial incidence of sudden death, total cardiac death and total mortality from all causes was 1%, 7% and 8%, respectively, at 1 year, and 4%, 24% and 26% at 5 years. The automatic implantable cardioverter-defibrillator nearly eliminates sudden death over a long-term follow-up period in a high risk group of patients. It has an acceptable rate of complications or problems, or both, and most late deaths in these patients are nonsudden and of cardiovascular origin.

Death, Sudden↗

Automatic implantable cardioverter-defibrillator: is early implantation cost-effective?

The evaluation of survivors of sudden cardiac death with serial electrophysiologic studies involves a lengthy and expensive hospitalization, especially when an automatic implantable cardioverter-defibrillator is ultimately necessary. The cost efficacy of this conventional approach was therefore compared with direct implantation of a cardioverter-defibrillator after the first electrophysiologic study. Thirty-two survivors of sudden death who had inducible ventricular tachycardia during their initial electrophysiologic study underwent serial drug trials. At discharge 12 (37%) were taking an antiarrhythmic drug found to prevent induction of ventricular tachycardia and 20 underwent cardioverter-defibrillator implantation after serial drug trials proved ineffective. The average length of hospitalization for this group that had undergone serial drug testing was 20.2 +/- 9.3 days at an average cost of $48,900 +/- $31,600. Seven survivors of sudden death had no inducible ventricular tachycardia during their initial electrophysiologic study and underwent direct cardioverter-defibrillator implantation. Their average length of hospitalization was 12.6 +/- 6.2 days at an average cost of $40,400 +/- $8,300. It is concluded that automatic implantable cardioverter-defibrillator implantation as an early intervention is not more costly and indeed may be cost-effective compared with therapy guided by serial electrophysiologic testing. As antitachycardia devices become more versatile, long lived and easier to implant, earlier implantation is likely to compare even more favorably with drug therapy.

Anti-Arrhythmia Agents↗

Sinus automaticity and sinoatrial conduction in severe symptomatic sick sinus syndrome.

Electrophysiologic studies with recordings of sinus node electrograms were performed in 38 patients with severe symptomatic sick sinus syndrome. Thirty-two of the 38 patients had episodic tachyarrhythmias and 17 presented with syncope. The clinically documented sinus or atrial pause was 5.6 +/- 2.8 s (mean +/- SD). Patients were divided into three groups according to electrophysiologic findings. Group I consisted of nine patients with complete sinoatrial block. Sinus node electrograms were recorded during the episodes of long pauses. Seven patients had unidirectional exit block, with the atrial impulse being capable of retrograde penetration to the sinus node causing suppression of sinus automaticity; two had bidirectional sinoatrial block. Group II consisted of 22 patients with either 1:1 sinoatrial conduction (group IIa = 13 patients) or second degree sinoatrial exit block (group IIb = 9 patients) during spontaneous sinus rhythm. Sinoatrial exit block, ranging from 1 to greater than 14 sinus beats, was observed during postpacing pauses that ranged from 1,650 to 37,000 ms (mean 7,286 +/- 6,989). The maximal sinus node recovery time ranged from 770 to 5,580 ms (mean 3,004 +/- 1,686) and was normal in 5 patients and prolonged in 17. Group III consisted of seven patients with no recordable sinus node electrogram, reflecting either a technical failure or a quiescence of sinus activity. The sinus node recovery time in these seven patients ranged from 1,190 to 4,260 ms (mean 2,949 +/- 1,121). Thus, abnormalities in both sinus node automaticity and sinoatrial conduction are responsible for the long sinus or atrial pauses in the sick sinus syndrome. However, complete sinoatrial exit block can occur and cause severe bradycardia with escape rhythm; repetitive sinoatrial exit block plays a major role in producing posttachycardia pauses.

Cardiac Pacing, Artificial↗

Automatic ventricular edge detection for determination of left ventricular volumes, ejection fraction and regional ejection fractions from first pass radioisotope angiography.

An automated method for detection of left ventricular (LV) outline (including the aortic and mitral valve planes) was developed for measurements of end-diastolic volume (EDV), end-systolic volume (ESV), global ejection fraction (EF), and regional EFs from first pass radioisotope ventriculography. The procedure includes: (1) interpolative background subtraction, (2) construction of gradient images, (3) automatic detection of valve planes, and (4) automatic radial search of LV outlines. The correlation between contrast angiography and Tc-99m first pass study was r = 0.84 (SEE = 23.4) for EDV, r = 0.93 (SEE = 12.8) for ESV, r = 0.84 (SEE = 6.91) for EF (via counts) and r = 0.80 (SEE = 8.56) for EF (via area-length method). Tests of intra-observer, inter-observer and inter-study variability revealed low level of variability. The results showed the potential of the automation of data processing for first pass radioisotope ventriculography.

Cardiac Volume↗

Automatic learning strategies and their application to electrophoresis analysis.

Automatic learning plays an important role in image analysis and pattern recognition. A taxonomy of automatic learning strategies is presented; this categorization is based on the amount of inferences the learning element must perform to bridge the gap between environmental and system knowledge representation level. Four main categories are identified and described: rote learning, learning by deduction, learning by induction, and learning by analogy. An application of learning by induction to medical image analysis is then exposed. It consists in the classification of two-dimensional gel electrophoretograms into meaningful distinct classes, as well in their conceptual description.

Electrophoresis, Gel, Two-Dimensional↗

Automatic segmentation of head MRI images by knowledge guided thresholding.

In this paper we present a procedure which automatically extracts soft-tissue from multi-sliced head MRI images, and then displays three-dimensional shapes of the extracted soft-tissues. We use an iterative thresholding algorithm for segmentation in which an optimum threshold value is decided based on a goodness measure we proposed. This procedure can be used in a preliminary diagnosis in brain surgery without much effort of users, because of whole procedure including threshold selection and segmentation is performed automatically for rendering a three-dimensional image of soft-tissue's surface on a graphic terminal.

Adipose Tissue↗

Automatic detection of irregular borders in melanoma and other skin tumors.

An irregularity index previously developed is applied to detect irregular borders automatically in skin tumor images, particularly malignant melanoma. The irregularity index is used to classify various tumor borders as irregular or regular. This procedure processes tumor images with borders automatically determined by a radial search algorithm previously described. Potential use of this algorithm in an in vivo skin cancer detection system and errors expected in the use of the algorithm are discussed.

Algorithms↗

An automatic color segmentation algorithm with application to identification of skin tumor borders.

A principal components transform algorithm for automatic color segmentation of images is described. This color segmentation algorithm was used to find tumor borders in six different color spaces including the original red, green, and blue (RGB) color space of the digitized image, the intensity/hue/saturation (IHS) transform, the spherical transform, chromaticity coordinates, the CIE transform and the uniform color transform designated CIE-LUV. Five hundred skin tumor images were separated into a training set and a test set for comparison of the different color spaces. Automatic induction was applied to dynamically determine the number of colors for segmentation. Ninety-one percent of image variance was contained in the image component along the principal axis (also containing the most image information). When compared to a luminance radial search method, the principal components color segmentation border method performed equally well by one measure and 10% better by another measure, including more near border points outside the tumor. The spherical transform provides the highest success rate and the chromaticity transform the lowest error rate, although large variances in the data preclude definitive statistical comparisons.

Algorithms↗

Automatic detection of intracranial contours in MR images.

Segmentation of the intracranial cavity in medical images is valuable in several research areas such as the quantitative analysis of normal and abnormal brain tissues, the registration of different imaging modalities (MRI, PET, CT) based on surface models of the brain, and the rendering of volume data. Because the manual delineation of the brain contour in the images can be demanding and error prone, an automatic procedure to perform this task is desirable. We have developed and tested a robust method that permits the automatic detection of the intracranial contour in transverse MR images. The method is described and its performance evaluated.

Algorithms↗

Methods of automatically acquiring images from digital medical systems.

Automated image acquisition plays an important role in a picture archiving and communication system (PACS). However, there is no single solution for automated data acquisition from existing digital medical imaging systems. We have gained a great deal of experience on automatic acquiring data by interfacing imaging scanners of major manufacturers. In this paper, we categorize the interface methods supported by the current image scanners. This categorization consists of five architectural models: (a) sequential chain; (b) direct interface; (c) memory access; (d) shared disk; and (e) interconnected network. The cost, rate of data transfer, and ease of implementation of each model are discussed. To ensure the integrity and availability of patient images in a PACS system, automated fault tolerance design in image acquisition is required. Based upon our field data, we report common scenarios which cause the acquisition to fail. We also describe techniques employed to automatically restart the operations which include recovery from acquisition processes' errors and traps, image acquisition computer down-time occurrence, and shutdown occurrence of medical imaging system.

Automation↗

Neurobiology of attention and automaticity.

Research in the field of attention and automaticity examines the nature of processing information with and without attention. Behavioral results show automatic processing is a continuous effect that is task specific and is affected by top-down influences. Neurobiological data show that differential attentional processing occurs in a distributed network, where initial performance is more strongly influenced by more anterior cortical structures. Recent models of attentional and autonomic processing directly incorporate physiological constraints and predict practice effects as a result of a variety of competing mechanisms.

Attention↗

A device for automatic measurement of writhing and its application to the assessment of analgesic agents.

A device was developed for automatically measuring writhing in mice so as to be applied to the assessment of analgesic agents. The device was composed of a specially designed container equipped with a detector, namely, a mechanoelectro transducer for writhing. The detector was made up of units of a string, two plates, and two strain gauges. In the unit, each end of the string was connected to either of the plates to which either of the strain gauges was attached. The change in tension of the string due to writhing was converted into the mechanical strain of the plates and then the resistance change of the strain gauges. The resistance change was amplified by a Wheatstone bridge circuit that was connected to a differential amplifier, a high-pass filter, comparator(s), and a monostable multivibrator to obtain the electrical signal for writhing. Using this device, writhing was continuously measured, and evaluation of various types of analgesic agents was performed. The result suggests that this device has sufficient accuracy both for the detection of writhing and the evaluation of analgesics. It has the advantage of automatic measurement of writhing in contrast to the conventional visual observation method.

Acetates↗

The development of an automatic method of safety monitoring at Pelican crossings.

This paper reports on the development of a method for automatic monitoring of safety at Pelican crossings. Historically, safety monitoring has typically been carried out using accident data, though given the rarity of such events it is difficult to quickly detect change in accident risk at a particular site. An alternative indicator sometimes used is traffic conflicts, though this data can be time consuming and expensive to collect. The method developed in this paper uses vehicle speeds and decelerations collected using standard in situ loops and tubes, to determine conflicts using vehicle decelerations and to assess the possibility of automatic safety monitoring at Pelican crossings. Information on signal settings, driver crossing behaviour, pedestrian crossing behaviour and delays, and pedestrian-vehicle conflicts was collected synchronously through a combination of direct observation, video analysis, and analysis of output from tube and loop detectors. Models were developed to predict safety, i.e. pedestrian-vehicle conflicts using vehicle speeds and decelerations.

Accidents, Traffic↗

Predicting severe head injury after light motor vehicle crashes: implications for automatic crash notification systems.

Motor vehicle crashes (MVC) are a leading public health problem. Improving notification times and the ability to predict which crashes will involve severe injuries may improve trauma system utilization. This study was undertaken to develop and validate a model to predict severe head injury following MVC using information readily incorporated into an automatic crash notification system. A cross-sectional study with derivation and validation sets was performed. The cohort was drawn from drivers of vehicles involved in MVC obtained from the National Automotive Sampling System (NASS). Independent multivariable predictors of severe head injury were identified. The model was able to stratify drivers according to their risk of severe head injury indicating its validity. The areas under the receiver-operating characteristic (ROC) curves were 0.7928 in the derivation set and 0.7940 in the validation set. We have developed a prediction model for head injury in MVC. As the development of automatic crash notification systems improves, models such as this one will be necessary to permit triage of what would be an overwhelming increase in crash notifications to pre-hospital responders.

Accidents, Traffic↗

Toward atlas-assisted automatic interpretation of MRI morphological brain scans in the presence of tumor.

RATIONALE AND OBJECTIVES: Determination of distorted brain anatomy surrounding a tumor causing a mass effect is much more difficult than interpretation of normal brain scans, particularly because this distortion is not easily predictable a tumor may be located in any place and vary substantially in size, shape, and radiological appearance. The objective of our work is to provide a qualitative means for rapid estimation of brain anatomy distorted by tumor. MATERIALS AND METHODS: Toward achieving this objective, we use an electronic and deformable brain atlas of gross anatomy along with a fast atlas-to-data warping technique. The deformed atlas determines the distorted anatomy surrounding a tumor and can be used for structure labeling (naming). The warping algorithm uses the Talairach transformation followed by three-dimensional nonlinear tumor deformation based on a geometric assumption that the tumor, delineated on radiological images, compresses its surrounding tissues radially. RESULTS: The approach is implemented and a dedicated application is developed. It processes the data automatically in five steps: (1) load data, (2) set the Talairach landmarks and perform the Talairach transformation, (3) segment the tumor, (4) warp the scan nonlinearly in three dimensions, and (5) explore the scan. The approach is very fast, and a magnetic resonance imaging scan is processed in 10-15 seconds on a standard personal computer. It is fully automatic and gives the user control over the entire process. CONCLUSION: Despite its limitations in modeling and validation, this practical solution provides a rapid and potentially useful qualitative assessment of anatomy deformed by a mass effect tumor.

Anatomy, Artistic↗

A deformable model for automatic CT liver extraction.

RATIONALE AND OBJECTIVES: This study was performed to design an automatic liver region extraction system to facilitate clinical liver size estimation and further serve as a prestage for liver reconstruction and volume estimation. MATERIALS AND METHODS: We present a modification of the well-known snakes algorithm for extracting liver regions in noisy CT images. Our modification addresses the issues of selection of the control points on an estimate of the contour and the determination of the weighting coefficients. The weighting coefficients are determined dynamically on the basis of the distance between the control points and the local curvature of the contour. RESULTS: The proposed method was used in extracting liver regions from 98 cross-sectional abdominal images. The overall performance was estimated by comparisons with original liver regions. CONCLUSION: The deformable model method enables an efficient and effective automatic liver region extraction in noisy environments. This approach eliminates human-in-the loop, which is the common practice for the majority of current methods.

Algorithms↗