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PubMed1995

[Image analysis for intercellular adhesion molecule-1 expression in MRI/lpr mice: effects of Chinese herb medicine].

To clarify the role of intercellular adhesion molecule-1 (ICAM-1) on pathogenesis of autoimmune lupus nephritis in MRL/lpr mice and the effects of Chinese herb medicine (stragalin), a computer image analysis system was used to study immunodepositions of ICAM-1, immunoglobulins and C3 in renal tissue sections of the mice. ICAM-1 was found in the mesangial area and deposited along the glomerular capillary walls in MRL/lpr mice. The distribution intensity of ICAM-1, immunoglobulins and C3 were significantly decreased after treatment with stragalin in form of decoction per os in the mice. We found that ICAM-1 might play an important role in pathogenesis of autoimmune lupus nephritis in MRL/lpr mice and Chinese herb medicine has some inhibition effects on immunodepositions in renal tissue of MRL/lpr mice.

Animals↗
PubMed2005

Image analysis supported moss cell disruption in photo-bioreactors.

Diverse methods for the disruption of cell entanglements and pellets of the moss Physcomitrella patens were tested in order to improve the homogeneity of suspension cultures. The morphological characterization of the moss was carried out by means of image analysis. Selected morphological parameters were defined and compared to the reduction of the carbon dioxide fixation, and the released pigments after cell disruption. The size control of the moss entanglements based on the rotor stator principle allowed a focused shear stress, avoiding a severe reduction in the photosynthesis. Batch cultures of P. patens in a 30.0-l pilot tubular photo-bioreactor with cell disruption showed no significant variation in growth rate and a delayed cell differentiation, when compared to undisrupted cultures. A highly controlled photoautotrophic culture of P. patens in a scalable photo-bioreactor was established, contributing to the development required for the future use of mosses as producers of relevant heterologous proteins.

Algorithms↗
PubMed1989

Neutrophil granulation stained with May Grunwald Giemsa or pure Azure B-eosin Y quantified by image analysis.

Neutrophil granulation was quantified after staining with May Grunwald Giemsa or with the pure dyes Azure B and eosin Y. Spinner slides of the buffy coat of 3 normal subjects and 14 persons with different grades of toxic granulation were studied. Morphometric parameters were measured using an image analysis computer (Texture Analysis System, Leitz Wetzlar, FRG). The parameters for granulation varied over a wider range in Azure B (AzB) than in May Grunwald Giemsa (MGG) stained granulocytes. This is in accordance with the visual microscopy observation, that granulation is more pronounced after staining with AzB than MGG. The predominant shade of the nucleus was similar in both stains, whereas considerable and variable differences in the shade of the cytoplasm were found.

Azure Stains↗
PubMed2000

A digital image analysis and neural network based system for identification of third-stage parasitic strongyle larvae from domestic animals.

A competitive learning vector quantization artificial neural network (ANN) was trained to identify third-stage parasitic strongyle larvae from domestic animals on the basis of quantitative data obtained from processed digital images of larvae. For this reason, various quantitative features obtained from processed digital images of larvae were tested as to whether they are variant or invariant to the shape taken by the motile larvae during image recording. A total of 255 images of 57 individual larvae in various shapes belonging to five genera were recorded. Following image processing, 16 features were measured, of which seven were selected as invariant to larva shape. By trial and error, two of those features, 'area' and 'perimeter', along with the quantitative features used in conventional identification, 'overall body length', 'width' and 'extension of sheath' (tip of larva to tip of sheath), were used as an effective training data set for the ANN. This ANN coupled with an image analysis facility and a knowledge relational database became the basis for developing a computer-based larva identification system whose overall identification performance was 91.9%. The advantages of this system are its speed and objectivity. The objectivity of the system is based on the fact that it is not subject to inter- and intra-observer variability arising from the user's profile of competency in interpreting subjective and non-quantifiable descriptions. The limitations of the system are that it cannot handle raw images but only data extracted from images, its performance depends on the reliability of the input vectors used as training data for the ANN, and its use is restricted only to well-equipped laboratories due to its requirement for expensive instrumentation.

Animals↗
PubMed1992

Prominent expression of type 1 (gamma) protein kinase C in Purkinje cells located in the "central mass" of Reeler mutant mouse cerebellum as revealed by a computer image analysis technique.

In Reeler mutant mice, cerebellar Purkinje cells exhibit abnormal synaptogenesis. In this study, Purkinje cells in the "central mass" with abnormal afferent connections were compared with those located in their normal position in the cortex in terms of immunoreactivity for type 1 (gamma) protein kinase C. A "computer image analysis technique" was developed for the purpose of this quantitative study, and it revealed that the immunoreactivity in the central mass was significantly higher than that in the cortex. The results suggest that type 1 (gamma) protein kinase C may be related to the abnormal synaptic formation of the Purkinje cells.

Animals↗
PubMed1997

Inhibition of elicitation of contact dermatitis in humans by mometasone furoate: evaluation by means of 20-MHz B scanning associated with image analysis.

BACKGROUND: Mometasone furoate (MF) is a new strongly lipophilic steroid which has an anti-inflammatory effect as evaluated by in vivo and in vitro studies. OBJECTIVE: The objective of the present study was to investigate the degree of inhibition of experimental allergic contact dermatitis induced by the treatment with MF. METHODS: The therapeutic effect was evaluated by an echographic method associated with image analysis. MF activity was compared to that of hydrocortisone acetate 0.5% (Cortaid Cream, Lachifarma), clobetasone butyrate 0.05% (Eumovate Cream, Glaxo) and clobetasol propionate 0.05% (Clobesol Cream, Glaxo), classified respectively as weak, moderately potent and very potent steroids. RESULTS: The different corticosteroid formulations employed for inhibiting experimentally induced contact dermatitis reflect the expected rank order of efficacy. CONCLUSION: MF behaves like clobetasol butyrate ranking as a moderately potent corticosteroid.

Administration, Topical↗
PubMed1998

Comparison of HPLC and stereologic image analysis for the quantitation of eu- and pheomelanins in nevus cells and stimulated melanoma cells.

The aim of the study was to compare two methods of quantitating eumelanins and pheomelanins, pigments synthesized by melanocytes. One is based on the high performance liquid chromatography quantitation of specific degradation products of each melanin type. The other requires image analysis, transmission electron microscopy, and stereology. In a previous study, we showed good correlations between both methods for total melanin but not for eumelanins or pheomelanins. We describe here the same comparison in more pigmented cells (nevus cells and stimulated HBL melanoma cells). Transmission electron microscopy micrographs were image analyzed to generate several primary parameters. Stereology was used for estimating melanosomal maturation, intracellular melanin content, and the number of melanized melanosomes per cell, for total melanin, eumelanins, or pheomelanins. Our results showed a good correlation between both methods for total melanin, eumelanins, and pheomelanins with an r equal to 0.99, 0.91, and 0.93, respectively, when all the points were used in the linear regression analyses. In the melanoma cell group (HBL cells cultured in media of different compositions), the chemical and morphometric estimations were not parallel in the case of eumelanins and pheomelanins. In addition, the stereologic and high performance liquid chromatography pheomelanins to eumelanins ratios were still not correlated. These results demonstrate the relevancy of the stereologic method, but the low level of melanization, the possible lack of specificity of melanogenesis in melanoma cells, and a problem of sensitivity of the stereologic method in this context seem to be obstacles in obtaining better results. The utilization of normal human melanocytes could give some answers to our hypotheses.

Chromatography, High Pressure Liquid↗
PubMed2006

Tissue specific differences in fibrillin microfibrils analysed using single particle image analysis.

Fibrillin microfibrils endow mammalian connective tissues with elasticity and play a fundamental role in the deposition of elastin. The microfibrils are 57 nm periodic supramolecular protein polymers with a mass of 2.5 MDa per repeat. The organisation of molecules within a microfibril is still open to debate and structural studies are only just starting to unravel this issue. The contribution of microfibril associated proteins to microfibril ultrastructure and whether there are any tissue specific differences in microfibril structure is still unknown. Therefore, we have used low dose electron microscopy, single particle image analysis and atomic force microscopy to study the structure of fibrillin microfibrils from different tissues. EM images of microfibrils from aorta, ciliary zonules and vitreous humor were collected and more than 500 microfibril repeats from each sample were subjected to averaging. Averages from each sample were analysed using axial stain exclusion patterns and difference images to detect any variations between them. The overall morphology of fibrillin microfibrils was conserved between tissues and there were only very minor differences in the bead and shoulder region of microfibrils. These data suggest that the structure of isolated microfibrils represents the fibrillin scaffold, and either microfibril associated molecules are lost on purification or play only a minor role in microfibril structure.

Animals↗
PubMed1995

Cytology, flow cytometry, image analysis, and interphase cytogenetics by fluorescence in situ hybridization in the diagnosis of transitional cell carcinoma in bladder washes: a comparative study.

The diagnosis of transitional cell carcinoma (TCC) in bladder washes is a diagnostic challenge to cytology. This study assessed the role of flow cytometry (FCM), image analysis (IA), and interphase cytogenetics by fluorescence in situ hybridization (FISH) as adjuncts in the cytodiagnosis of TCC in bladder washes. Forty separate samples of bladder washes were prospectively evaluated by conventional cytology (CY), FCM, IA, and FISH, and the results were compared with the subsequent surgical biopsy specimens which revealed 26 TCC (3 GR I, 6 GR II, 17 GR III) and 14 benign lesions. Using histology as the "gold standard" and following the previously published criteria for detection of TCC by CY, FCM, IA, and FISH, the concordance rates between histology and CY, FCM, IA, and FISH were 75, 74, 89, and 83%, respectively. CY, FCM, IA, FISH, and histology were concordant in 54% of the cases. The sensitivity of CY, FCM, IA, and FISH were 61, 72, 91, and 73%, respectively, while the specificity were 100, 80, 83, and 100%, respectively. The combined sensitivity of all the parameters was 96%. Interestingly, the false positive cases by FCM and IA showed cystitis. We conclude that IA has the highest sensitivity in detecting TCC in bladder washes followed by FISH, FCM, and CY, while CY and FISH have the highest specificity. This study indicates that FCM, IA, and FISH are useful adjuncts to cytology in the diagnosis of TCC in bladder washes. The finding of DNA-aneuploidy in cystitis warrants further investigation.

Carcinoma, Transitional Cell↗
PubMed2004

An automated scoring procedure for the micronucleus test by image analysis.

The micronucleus assay (MNT) in human lymphocytes is frequently used to assess chromosomal damage as a consequence of environmental mutagen exposure, to assess the effect of mutagens or to search for reduced DNA repair capacity after a mutagenic challenge. We have established an automated scoring procedure for the cytokinesis blocked MNT based on computerized image analysis (Metasystems Metafer 4 version 2.12). To evaluate the results we used the reproducibility of counts, established a dose-response curve for gamma-irradiation and used the ability of the system to differentiate between breast cancer patients and controls as a biological reference, a difference which we had observed before by visual counting. Blood cultures were irradiated with gamma-rays (2 Gy) at the beginning and treated with cytochalasin B during the last 24 h. The slides were stained with Giemsa for visual counting and with DAPI for automated analysis. Our test sample consisted of 73 persons (27 with breast cancer and 26 female and 20 male controls). A comparison between visual counting (controls, mean MN frequency 313) and automated counting (mean MN frequency 106) in slides from the same culture revealed a large drop for the automated counts. However, the automated counts were as reproducible as the visual counts [coefficient of variation (CV) on the sample approximately 20%; CV on repeated counts of the same slides approximately 5%] and both counts were highly correlated. Furthermore, the discrimination between cases and controls improved for automated counting of slides from the same cultures [visual odds rato (OR) < or = 4.0, P = 0.009; automated OR > 16, P < 0.0001], with a strong dependence on the set of parameters used. This improvement was confirmed in a validation sample of an additional 21 controls and 20 cases (OR = 11, P = 0.0018) performed as a prospective or diagnostic test.

Algorithms↗
PubMedDate not supplied

The combination of the tetrazolium derivative reduction (MTT) and digital cell image analysis to monitor in vitro the cytotoxicity of anti-neoplastic drugs.

We assessed the in vitro drug-induced cytotoxicity by means of the rapid low-cost but weakly sensitive Thiazolyl blue (MTT) test, and the less rapid, higher cost, but highly sensitive cell image analysis (CIA) test. We studied the influence of three drugs, i.e. a vinca-alkaloid (Navelbine), an alkylating investigational agent (PE1001), and an intercalating drug, i.e. adriamycin, on the proliferation (MTT test) and cell kinetics (CIA test) of the mouse MXT and the MCF-7 mammary cancer cell lines. Adriamycin and PE1001 decreased MXT and MCF-7 cell proliferation in a dose-dependent manner as assessed by both the MTT and CIA tests. Navelbine was highly cytotoxic in a dose-dependent manner. We demonstrated that Navelbine arrests cells in the M phase without altering the G2 phase. In sharp contrast, PE1001 arrest cells in the G2 phase without altering the M phase. An adriamycin-induced effect was apparent on S phase. Thus, the MTT test allows the screening of a great number of drugs analyzed at the cell proliferation level and, in the case of MTT positive drug-induced response, the CIA test enabled the drug-induced effect at cell cycle kinetic level to be investigated.

Adenocarcinoma↗
PubMed1994

[Modern image analysis processes as support for spatial reconstruction of histological structures].

Spatial reconstruction of anatomic and histologic structures is of increasing importance in biological and medical research. A special kind of disease--hereditary opalescent dentine (HOD)--offered a good chance for the use of a comfortable image-analysis system (IBAS, Kontron) in combination with a special software for three-dimensional reconstruction. New aspects of the morphologic organisation of a key structure of HOD--the giant dentinal canals--could be shown, thus supporting the hypothesis of a vasal origin.

Adolescent↗
PubMedDate not supplied

Adrenal cytomegaly: quantitative assessment by image analysis.

Adrenal cytomegaly was found in 23 or 0.8% of 2711 pediatric autopsies, including a 13-year-old girl with leukemia and a 20-year-old woman with cystic fibrosis. Cytomegaly of pancreatic islet and acinar cells and pituitary cells was also noted. DNA quantitation by image analysis indicates that cytomegalic cells of the adrenal cortex, pancreas, and pituitary contain increased amounts of DNA (hyperdiploid aneuploid), with adenocytes containing over 25 times the normal amount of DNA in some instances. These data and morphological features suggest that cytomegaly is a reflection of polyploidy due to either partial DNA replication or partial DNA replication plus other modes of polyploidization. Although the mechanism(s) by which these changes evolve remains unknown, regressive processes do not seem to be directly involved. We speculate that polyploidization is, in this setting, a response to physiological demand, a concept that also has relevance to the interpretation of morphology and DNA ploidy profiles of endocrine tumors.

Adolescent↗
PubMed1994

Differential intracellular processing of the anthracycline drug ME2303 in doxorubicin-sensitive (A2780) and -resistant (A2780AD) human ovarian cancer cells as studied with confocal laser scanning microscopy and image analysis.

Laser scanning confocal microscopy has been used to follow the uptake and efflux of the 2-fluoroglycoside of doxorubicin, ME2303, in live cultures of the human ovarian cancer cell line A2780 and its doxorubicin-resistant variant A2780AD. Our methods combine confocal laser scanning microscopy and image analysis to examine the dynamics of anthracycline drugs in cancer cells. Cytotoxicity determined by MTT dye reduction showed that A2780AD cells were more than 400 times less sensitive to doxorubicin compared to A2780 cells but almost 9 times more sensitive to ME2303 compared to doxorubicin. The naturally fluorescent drug was tracked within live cells at 37 degrees C to provide time-course information in relation to nuclear and Golgi-associated cellular domains, as indicated by BODIPY FL ceramide-associated fluorescence. In both cell types, ME2303 was characterised by strong nuclear membrane and perinucleolar fluorescence and as a localised punctate pattern within the nucleus. A2780AD cells accumulated ME2303 in their nuclei at a much reduced rate compared to the doxorubicin-sensitive cells, and ME2303 efflux from resistant cell nuclei was approximately twice as fast as from A2780 cells. The relative uptake of ME2303 into Golgi-associated domains and the nucleus were monitored simultaneously during the initial 35 min of exposure to 10 microM ME2303 and during the first 45 min of a "chase" culture following exposure to 20 microM ME2303. ME2303 was detectable within the Golgi-associated domains of A2780 cells several minutes later and in less relative concentration than in A2780AD cells 15 min into a chase culture. Our results suggest the direct involvement of differences in drug processing via the Golgi apparatus in the expression of P-glycoprotein-related drug resistance.

Coloring Agents↗
PubMed1992

Bradykinin-induced Ca2+ entry, release, and refilling of intracellular Ca2+ stores. Relationships revealed by image analysis of individual human fibroblasts.

Lys-Bradykinin (BK), a mitogen for human foreskin fibroblasts (HSWP cells) (Owen, N. E., and Villereal, M. L. (1983) Cell 32, 979-985), elicits a rapid, transient elevation of intracellular free Ca2+ concentration ([Ca2+]i) in these cells. We have used image analysis of fura-2-loaded HSWP cells to examine the BK-induced [Ca2+]i changes in individual cells. BK-stimulated Ca2+ entry and release of intracellular Ca2+ stores can be distinguished by stimulating cells in the presence or absence of extracellular Ca2+, or by inhibiting Ca2+ entry with 5 mM NiCl2. BK-sensitive intracellular Ca2+ stores can be depleted by exposure of the cells to BK in Ca(2+)-free medium; refilling of the stores requires extracellular Ca2+. A component of BK-stimulated Ca2+ entry persists after removal of agonist, but inactivates with a t1/2 of approximately 5 min. Although previous studies have attributed the Ca2+ entry which persists after agonist removal to a "capacitative Ca2+ entry" pathway activated by the depletion of the intracellular Ca2+ stores, we find that a large component of this BK-stimulated Ca2+ entry is not due to capacitative Ca2+ entry since (1) ionomycin can deplete the BK-sensitive intracellular Ca2+ stores without appreciably stimulating Ca2+ entry and without inhibiting the BK-stimulated Ca2+ entry and (2) this Ca2+ entry pathway inactivates at a time when the Ca2+ pools are still empty and a capacitance entry pathway should still be open. On the other hand, refilling of the intracellular Ca2+ stores can occur after the noncapacitative Ca2+ entry component has inactivated or when it is inhibited by Ni2+; in these cases refilling occurs without a detectable elevation of [Ca2+]i suggesting that refilling of internal Ca2+ pools might occur by a capacitative route.

Biological Transport↗
PubMed2002

[Quality assurance of powders for bioceramic applications with image analysis].

Bone substitute materials are very demanding with respect to their quality, in particular, a high toughness and durability are required. In case of producing the composites by sintering of mechanically generated powder mixtures, a high efficiency of mixture is demanded, too. The characterisation of such particle systems is difficult. An effective method is the image analysis supported by statistical evaluation of both the size and the shape distributions of particles and agglomerates. As it is proved by first results, powder mixtures of hydroxyapatite can be described by the geometry of voids in the agglomerate structure using the shape factor weighted by area of the respective image object. As a measure of the efficiency of mixture, the degree of reproducibility was introduced.

Bone Substitutes↗
PubMedDate not supplied

Micro-Raman imaging analysis of monomer/mineral distribution in intertubular region of adhesive/dentin interfaces.

It is generally proposed that bonding of resins to dentin results from infiltration of the adhesive monomers into the superficially demineralized dentin. However, it is still not clear how well the mineral phase of dentin is removed and how far each monomer penetrates into the thin zone of "wet" demineralized dentin. The quality and molecular structure of adhesive/dentin interfaces formed under "wet" bonding conditions are studied using 2-D Raman microspectroscopic mapping/imaging techniques. Micro-Raman imaging analysis of the adhesive/dentin interface provides a reliable and powerful means of identifying the degree and depth of dentin demineralization, adhesive monomer distribution, and flaws or defects in the pattern of adhesive penetration. The image of mineral reveals a partially demineralized layer on the top of dentin substrate. Adhesive monomers readily penetrate into dentin tubules and spread into intertubular region through open tubules. The extent of adhesive monomer penetration is higher in the intertubular regions close to tubules as compared to the middle regions between the tubules. The diffusion of resin monomers differs substantially. In a comparison with a hydrophilic monomer, the hydrophobic monomer resists diffusion into the demineralized intertubular dentin area.

Absorption↗
PubMed2006

A computer-based, image-analysis method to quantify HIV-1 infection in a single-cycle infectious center assay.

Rapid and precise quantitation of the infectivity of HIV is important for molecular virologic studies as well as for measuring the activities of antiviral drugs and neutralizing antibodies. In the present study, an indicator cell line and image-analysis software were used to quantify HIV infectivity. Cells of the P4R5 line, which express the receptors for HIV infection as well as beta-galactosidase under the control of the HIV-1 long terminal repeat, were infected either with CXCR4- or CCR5-using viruses, including primary isolates, then stained 2 days later with X-gal to turn infected cells blue. Digital images of monolayers of the infected cells were captured using a high resolution CCD video camera and a macro video zoom lens. A software program was developed to process the images and to count the blue-stained foci of infection. The assay was applied to assess the infectivity of site-directed viral mutants, and to measure the activity of antiviral drugs and neutralizing antibody. The results indicate that the described method allows for the rapid quantitation of infected cells over a wide range of viral inocula with reproducibility, accuracy and relatively low cost.

Anti-HIV Agents↗