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Image analysis as a monitoring tool for activated sludge properties in lab-scale installations.

An important step in the battle against filamentous bulking is the development of a monitoring system for activated sludge properties. Therefore, a fully automatic image analysis method for recognizing and characterizing flocs and filaments in activated sludge images has been developed. This procedure has been subsequently used to monitor activated sludge properties in a lab-scale installation. The results of a 100-days experiment indicate that the image information correlates well with the evolution of standard settling properties, in this case the Sludge Volume Index. It is shown that, at the onset of severe filamentous bulking, there is an increase in total filament length on the one hand, and a significant change in floc shape on the other hand.

Automation↗

Grading of precancerous laryngeal lesions by multiparameter image analysis at separate epithelial layers.

In order to detect early precancerous changes which do not involve the whole thickness of the epithelium, we used a novel image analysing program based on an IBAS system (Kontron, Germany) to determine nuclear DNA content (NC) as well as average nuclear area (NA) and variation of nuclear area (VA), in the entire epithelium and in three sublayers, parabasal, intermediate, and superficial. DNA aneuploidy was found in only half of the cases classified as 'high-grade' (HG) lesions, comprising carcinoma in situ (CIS) and severe epithelial dysplasias (EDIII), and was chiefly demonstrable in the parabasal third of the epithelium. The other lesions were DNA euploid. HG lesions showed highly significant increases of NA and VA at the lower levels of the epithelium when compared with 'low-grade' (LG) lesions comprising moderate and mild epithelial dysplasias (EDII and EDI). Our data show that the combination of multiparameter image analysis with conventional morphology assists in the objective grading of precancerous lesions and permits the reliable detection of high-grade lesions.

Adult↗

Comparison of analog and digital Fourier transforms in medical image analysis.

The effectiveness and limitations of medical image processing using analog and digital methods are studied. Several types of errors introduced during the image processing are analyzed. For the analog optical Fourier transform, errors are introduced by the vignetting effect and lens aberration. For the digital Fourier transform, errors are introduced by the aliasing effect and the band limit. To compare the results obtained by the two techniques, a set of x-ray images was processed both optically and digitally. The former was achieved by an optical system containing a large Fourier telephoto lens and the latter by a personal computer using a Fourier transform algorithm. The veracity of both the optical and digital Fourier spectra is analyzed. Our results indicate that the optical method has high speed due to parallel processing. High veracity can be achieved in high frequency regions by using an optimal optical system. In comparison, the digital method has the advantages of high processing precision and programmability, but has low processing speed. The comparison of the two different techniques presented in this article can provide a basis for selection of the processing method in different clinical settings. Even with today's fast computers, the optical method is still suitable for many clinical applications. The best choice lies in an analog-digital combination.

Algorithms↗

Measurement of iris color using computerized image analysis.

PURPOSE: To develop a standardized method for measuring from iris photographs, light and dark segments of areas and densities of iris color. METHODS: Computerized image analysis was used to measure the iris photographs. The reproducibility of this method was studied in 30 normal eyes with three different colored irides, green-brown, blue-grey brown and yellow-brown. Three photographs were taken of each iris with a slit lamp camera at three different exposures at baseline. The photographs were repeated with exposure providing for the best reproducibility at 6.5 +/- 1.7 months as a first follow-up after baseline and 3.6 +/- 0.8 months following the first follow-up visit. At least one measurement was made for each photograph. RESULTS: The mean percent coefficient of variation (standard deviation of triplicate measurements/mean x 100) ranged from 1.0 to 4.1% for area and density measurements. Furthermore, the range of mean percent differences between baseline and follow-up visits ranged from 1.2 to 6.3%. CONCLUSION: We have developed a standardized method which appears suitable for measuring changes over time in iris color.

Aged↗

Effects of noise and quinine on the vessels of the stria vascularis: an image analysis study.

Surface preparations of the stria vascularis from guinea pigs exposed to wide-band noise or intoxicated with quinine monohydrochloride dihydrate were studied by light microscopy and computerized image analysis in order to evaluate quantitatively the effects of these agents on two characteristics of the strial vasculature: vascular density and erythrocyte distribution. An image analyzer was used to measure the area of strial vessel lumen and erythrocyte distribution as a fraction of the total area of strial tissue under observation. The results demonstrate that changes in the strial vessels do occur in guinea pigs exposed to noise or given large doses of quinine. Localized vessel narrowings caused by swollen endothelial cells and possibly by contraction of pericytes were found in both experimental groups, but there was no apparent tonotopical relationship between these effects and the reduction in cochlear potentials. A significant reduction in the number of erythrocytes was found in all turns of the cochlea in both experimental groups. Although a significant difference in vascular density was found among turns of the cochlea in both experimental and control animals, there was no widespread change in vascular density as a result of either noise exposure or quinine treatment.

Animals↗

Automated facial image analysis: detecting improvement in abnormal facial movement after treatment with botulinum toxin A.

The purpose of this study was to evaluate the ability of Automated Facial Image Analysis (AFA) to detect changes in facial motion after Botox injections in patients with facial nerve disorders accompanied by abnormal muscle activity. Eight subjects received Botox for oral to ocular synkinesis (n = 6), ocular to oral synkinesis (n = 1), and/or depressor anguli oris overactivity (n = 3). Subjects were video-recorded during 2 directed facial action tasks before and after Botox treatment. AFA measurement and Facial Grading System (FGS) scores were used to evaluate the effects of Botox. After Botox, AFA detected a decrease in abnormal movements of the eyelids in all patients with oral to ocular synkinesis, a decrease in oral commissure movement for the patients with ocular to oral synkinesis, and an increase in oral commissure movement in all patients with depressor overactivity. The FGS scores failed to demonstrate any change in facial movement for the case of ocular to oral synkinesis and for 2 cases of depressor overactivity. AFA enables recognition of subtle changes in facial movement that may not be adequately measured by observer based ratings of facial function.

Adult↗

Twofold symmetry of human fibrinogen proved at the beta chain distal domains by monoclonal-immunoelectron microscopy and image analysis.

Using a murine antibody (F7) specific for the C-terminal domain of the beta chain of human fibrinogen combined with electron microscopy and image analysis, we show unequivocally that the epitopes for F7 are at the distal nodules of fibrinogen, equidistant from the center of the molecule and arranged not colinearly with the long axis of the molecule but at opposite sides of it, i.e., following twofold symmetry. Thus, given the monoclonality of the immunochemical probe used and the dimeric nature of the fibrinogen molecule, we can conclude that the distal domains of the two beta chains are arranged in the same manner as these epitopes and, therefore, that the fibrinogen molecule has twofold symmetry. This symmetry pattern found here for F7 is the same as that found recently for the platelet fibrinogen receptor binding sites [Weisel, J. W., Nagaswami, C., Vilaire, G., & Bennett, J. S. (1992) J. Biol. Chem. 23, 16637-16643], located almost certainly at the C-terminal end of the gamma chains, and gives further support to the most accurate model of fibrinogen available so far. We discuss the consequences of this symmetry pattern and of the molecular rigidity of fibrinogen in the actual models of fibrin polymerization and platelet aggregation and adhesion.

Amino Acid Sequence↗

Density and phenotype of tumour-associated mononuclear cells in colonic carcinomas determined by computer-assisted video image analysis.

The density and phenotypes of tumour-associated mononuclear cells (TAMC) in tissue sections of colonic carcinomas was determined by the technique of video image analysis (VIA). This technique allowed an accurate and objective enumeration of both total mononuclear cells (MC) in H&E stained sections and individual types of cells as revealed by immunoperoxidase staining with monoclonal antibodies in frozen sections. This enumeration allowed reliable statistical analysis of the differences between sample groups. Using this technique it was found that the density of MC in histiologically normal tissue was significantly higher than in tumour tissue. Tumours from patients with the best prognosis (stage A) had significantly higher numbers of TAMC than stage B (P less than 0.02), C (P less than 0.002) and D (P less than 0.002) tumours. The differences in the density of TAMC between tumours obtained from stage B and C and that between C and D were not significant, whereas stage B had a significantly higher TAMC density than stage D tumours (P less than 0.05). Comparing tumour differentiation, well differentiated adenocarcinomas had a significantly higher (P less than 0.05) TAMC density than poorly differentiated tumours but not moderately differentiated tumours. Moderately and poorly differentiated adenocarcinomas did not differ significantly in the density of TAMC. In examining the phenotype of these cells, it was found that T lymphocytes formed the majority of the TAMC with the CD4+ subset predominating in 28 of 29 cases. Similarly, all sections of normal colon (taken at least 4 cm away from the tumour) had more CD4+ than CD8+ cells. The proportion of the total leucocyte population that was CD3+ was comparable in normal and tumour tissue. Generally, few macrophages were present in either tumour or normal tissues. B cells (CD21%) and subset of NK cells (CD57+) were not detected in the tumours. There were no significant differences in the proportion of leucocytes which were CD4+, CD8+ and CD14+ (macrophages) between the normal colon and the tumour tissues. The types of cells in the TAMC population did not differ with tumour stage or differentiation or with the density of the TAMC itself.

Adenocarcinoma↗

Automated monitoring of cell concentration and viability using an image analysis system.

In order to automate measurements of cell concentration and viability in a suspended animal cell culture, we have developed an in situ microscopic image analysis system with an effective cell recognition algorithm. With a small amount of sample, this system can measure the cell density rapidly and aseptically. In addition, it can measure a cell size histogram including cell debris small particle distribution. These small particles have been found to be related to the viability of the mouse-mouse hybridoma STK1 cell line. By using cell debris small particle density as an indicator of cell viability, the developed system provides non-destructive viability monitoring without trypan blue staining.

Animals↗

Structure of an influenza neuraminidase-diabody complex by electron cryomicroscopy and image analysis.

The structure of a complex between a bivalent diabody and its antigen, influenza neuraminidase, has been determined by electron cryomicroscopy of single particles and image analysis. A three-dimensional reconstruction has been interpreted in terms of high-resolution X-ray models of the component proteins. The complex consists of two neuraminidase tetramers cross-linked by four diabodies with 422 point symmetry. The structure and symmetry of the complex is determined uniquely by packing constraints consistent with the maximum possible number of diabody cross-links. Diabodies may provide a useful approach to the structure determination of small proteins by incorporating the proteins into large symmetric complexes followed by single-particle electron microscopy.

Cryoelectron Microscopy↗

Hairlessness does not influence growth retardation in nude mice. Computer image analysis study.

Ossification in 4-week-old nu/nu and nu/+ BALB/c and BFU mice was studied by X-ray analysis and by measurement of the thickness of the proximal tibial growth cartilage using CUE 4 Olympus computer image analysis. Not only altered architecture but also a significantly thinner proximal tibial growth plate was observed in athymic nu/nu as opposed to nu/+ and BFU mice. On the other hand, no significant differences were found between nu/+ and BFU littermates. Higher X-ray density of tail vertebrae was observed in nu/+ and BFU than in nu/nu mice. This comparison between athymic nu/nu and hairless euthymic BFU mice indicates that altered postnatal ossification in nude mice is not caused by hairlessness, but is due to other (immunological or endocrinological) differences.

Animals↗

Quantification of proliferating cell nuclear antigen in large intestinal crypt by computer-assisted image analysis.

Measurements of proliferating cell nuclear antigen (PCNA) labeling of the large intestinal crypts scored by experienced observers were compared with those generated by computer-assisted image analysis (CAIA). (CAIA was performed at Pathology Expertise, Inc., Newton, MA, by A. V. Sotnikov.) Serial sections (3 microns) of the rectal biopsy specimens from 32 patients were immunostained for PCNA and then counterstained with hematoxylin. The same set of slides and rules for the location/acquisition of complete crypts was used to assess a minimum of ten complete crypts/patient. Each crypt was subdivided longitudinally into five equal compartments. With CAIA, the images were stored digitally, and once the color references were set, the areas occupied by labeled and counterstained nuclei were quantified automatically. The labeling index (LI) was calculated from the PCNA-labeled nuclei area/total nuclei area in CAIA and from the number of labeled cells/total number of cells in visual scoring. The LI of whole crypts averaged 1.04 +/- 0.18 by CAIA and 3.91 +/- 0.46 by the visual method, and the Spearman correlation (rs) between the two methods was 0.89. The different modes of evaluation and color reference selection are likely to have contributed to the differences in the LI ranges observed in the two methods. The high correlation between PCNA quantification by CAIA and visual scoring by experienced technicians indicates that CAIA can reliably rank individual subjects. Thus, measurement of PCNA labeling by CAIA is a practical alternative for evaluating colorectal epithelial cell proliferation.

Adult↗

Improved algorithms for automatic bone histomorphometry on a numerized image analysis system.

Quantitative histological methods have proved to be the most effective methods in bone disease research. Faster and more accurate techniques are currently needed. We have developed a simple digitized image analysis system which allows accurate measurements of trabecular bone mass. The algorithm is based on the 'four-connected sets' mathematical theory. Given a numerized image displayed by a CCD camera, the algorithm recognizes all possible four-connected sets and provides area measurements. The first procedure automatically eliminates small, irrelevant profiles (wrinkles, cell nuclei, etc.) while larger profiles are erased interactively. The second procedure similarly erases the artefactual defects within the trabeculae (artefactual cracks or empty osteocytic lacunae). The method was shown to be very accurate and time-saving.

Algorithms↗

Detection of aneuploidy and polyploidy in non-Hodgkin's lymphoma by computer video image analysis.

Thirty-six cases of histologically intermediate and low grade non-Hodgkin's lymphoma, classified according to the International Working Formulation, were examined using image analysis cytometry. Aneuploidy was detected in 21 cases (58%). No significant difference in either the incidence or extent of aneuploidy was seen for cases that entered remission when compared to cases that did not. Bone marrow examinations, by either aspiration or trephine biopsy, were conducted on 34 cases, with 11 showing lymphocytic infiltration. Aneuploidy was detected in 6 of the 11 cases having bone marrow involvement, while 5 cases with bone marrow infiltration were euploid. Polyploid cell distributions were detected in 3 cases, and the remaining 12 cases were diploid. The biologic significance of polyploidy among non-Hodgkin's lymphomas is not well understood, and its detection is limited to static techniques for DNA quantitation. With further investigation, awareness of its presence may reveal some insight into the clonal evolution of this neoplasm.

Adolescent↗

Evaluation of a simple method for measuring the cellular DNA content of mouse oocytes and embryos, human fibroblasts and parthenogenetically activated human oocytes using a computerised image analysis system (Seescan).

We report the use of a simple, reproducible, photocytometric method for measuring nuclear DNA content of DAPI-stained cells, using a computerised image analysis system: Seescan. As this technique is non-destructive and uses very short exposure to ultraviolet light, it can be used for either fixed or vital material. After correcting for any background cytoplasmic staining, the intensity of nuclear stain was measured by the Seescan and compared with that of control cells of known ploidy. Fixed material was found to stain more intensely than live material initially, but demonstrated a rapid loss of nuclear intensity over the first 90 min following removal from DAPI, after which the level plateaued. In contrast, live cells showed no change in nuclear intensity with time. The system was validated by measuring the DNA content of carefully timed mouse blastomeres, human fetal lung fibroblasts and parthenogenetically activated human oocytes. The results obtained were appropriate for the developmental stage or phenotypic appearance of each of the cell types measured.

Animals↗

Use of computerized image analysis to quantify staphylococcal adhesion to canine corneocytes: does breed and body site have any relevance to the pathogenesis of pyoderma?

An optimized system of computerized image analysis was used to investigate variations in the adherence of Staphylococcus intermedius to canine corneocytes from four different breed groups and six different anatomical sites. S. intermedius showed significantly greater adherence to the head and neck compared with the dorsum, but adherence to the limb, axilla and groin did not differ from other sites. Furthermore, there was significantly greater adherence of S. intermedius to corneocytes from the dorsum, forelimb, axilla and groin of Boxers and Bull Terriers than Spaniels and Hounds. S. intermedius, and also Pseudomonas aeruginosa, exhibited abundant adherence, which was significantly greater than Staphylococcus aureus, Streptococcus canis, Klebsiella pneumoniae and Escherichia coli. In addition, S. intermedius adherence demonstrated a sigmoid dose-response curve with increasing bacterial concentration. These results suggest that S. intermedius adheres to canine corneocytes by a specific receptor-ligand interaction and adheres to the skin of some breeds more avidly than others. However, variations in adherence between body regions would not account for the predilection sites of canine bacterial pyoderma.

Animals↗