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Biomedical subjects

K Ostrowski

Publications and source records attributed to K Ostrowski.

At least 91 records · Page 5Linked to original sources
PubMed1986

Fourier analysis of the cell shape of paired human urothelial cell lines of the same origin but of different grades of transformation.

The rationale of the present investigation is the observations made by many authors of changes in the molecular structure of the cell surface during the multistep process of malignant transformation. These changes may influence cell-matrix and cell-cell interactions and thereby cause changes in cell adhesiveness and cell shape. The aim of the present work was to investigate whether the development of various grades of transformation in vivo and in vitro of human urothelial cells is accompanied by significant changes in cell shape as measured by Fourier analysis. The following transformation grades (TGr) have been defined (Christensen et al. 1984; Kieler 1984): TGr I = nonmalignant, mortal cell lines that grow independently of fibroblasts and have a prolonged life span. TGr II = nonmalignant cell lines with an infinite life span. TGr III = malignant and immortal cell lines that grow invasively in co-cultures with embryonic chick heart fragments and possess tumorigenic properties after s.c. injection into nude mice. Comparisons of 4 pairs of cell lines were performed; each pair was of the same origin. Two pairs--each including a TGr I cell line (Hu 961b and Hu 1703S) compared to a TGr III cell line (Hu 961a or Hu 1703He)--were derived from two transitional cell carcinomas (TCC) containing a heterogeneous cell population. Two additional cell lines classified as TGr II (HCV-29 and Hu 609) were compared to two TGr III sublines (HCV-29T and Hu 609T, respectively) which arose by "spontaneous" transformation during propagation in vitro of the respective maternal TGr II-cell lines.(ABSTRACT TRUNCATED AT 250 WORDS)

Carcinoma, Transitional Cell↗
PubMed1986

The application of flow cytophotometry in measurements of cell adhesion.

A common approach to the study of cell substrate interactions is the measurement of the attachment of cells to different substrates or to cultured cell layers. The evaluation of attachment is made either by scintillation counting of previously labelled adhering cells, or by light microscopy using the criterion of cell shape, sometimes refined by automatic image analysis. These methods have many drawbacks. This paper suggests the use of fluorescence-activated flow cytophotometry, (FC) which yields direct counts of the non-adhering cells. These "free" cells are removed after completion of the adhesion experiment from the microtitre plate wells. An internal standard, in the form of fluorescent polystyrene beads is added, allowing evaluation of the percentage of cells adhering to the well walls. Flow cytophotometry then produces data based on the analysis of large populations of cells. Unequivocal discrimination is obtained between the counted cells and counted fluorescent beads eliminating counting errors. The results can be processed on line by computer. A suspension of mouse splenocytes was used for the evaluation of the overall error of the method arising from inaccuracies in pipetting, interference of glutaraldehyde with ethidium bromide (EB) staining and instrumental error. Each adhesion experiment was terminated by staining and post-fixation and it was established that this introduces no change in cell counting, in comparison with the original unfixed cells. Prefixation, however, quenches the EB staining and would interfere with the counting procedure. The overall standard error of the technique was found to be 5%-10%.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1986

Objective evaluation of fibrosis in human testicular biopsies by analysis based on optical diffractometry.

Bilateral testicular biopsies were obtained from 27 patients and submitted to two different treatments for each of them: firstly, a classical fixation and colouring, permitting the histopathological diagnosis and subjective appreciation of the degree of fibrosis; secondly, a new objective technique based on optical diffractometry: this method is based on the analysis of light intensity distribution in the diffraction pattern of an original image. Using two different methods of discriminant analysis, we observed some errors due to the subjective examination; in particular, several fibrosed samples had been judged as 'normal' by the histopathologist. Moreover, we showed the existence of some heterogeneity between different slides from the same original biopsy.

Biopsy↗
PubMed1984

Fourier analysis of the shape of normal and transformed epithelial cells derived from human transitional epithelium.

The aim of this paper is to show the possibility of objective mathematical description of changes occurring in the shape of cells in the process of transformation. The evaluation of the changes in cell shape of the chosen cell lines differing in transformation grade was performed by the use of Fourier analysis of the shape. Any two-dimensional contour can be described with specific accuracy in a mathematical manner using the closed form Fourier series of cosines. The components forming the analysed shape, called harmonics, are independent and uncorrelated measures of their contribution to the total shape. The shape of each cell can be represented by the spectrum of harmonic amplitudes. To quote the paper by Healy-Williams and Williams (1981): "The observed shape is partitioned into series, where gross shape, as elongation or triangularity, is measured by the harmonic amplitudes of the lower harmonic order and increasingly fine scaled surface sculpture is measured at higher orders". The statistically evaluated results allow the objective comparison of the cell shapes of several compared cell lines differing in transformation grades. Malignant transformation is supposed to be a multistep process. The different grades of transformation could be defined by several parameters as changes in the morphology of the cells, their ability to compete with fibroblasts, their life span, their angiogenic potency, their invasiveness in vitro and their tumorigenicity in nude mice. In this paper several human urothelial cell lines of normal and tumor origin differing in their transformation grade (TGr I-III) were compared by the use of Fourier analysis of their shape. TGr I cultures have finite life span but do not need intermittent collagenase treatment to prevent fibroblast overgrowth. TGr II cultures acquire infinite growth potential, here defined as capacity to survive at least 70 passages. They are neither tumorigenic nor invasive. TGr III cultures show infinite growth transformation, increased angiogenicity and ability to invade normal host tissue in vitro. They produce progressively growing tumors in nude mice. The following human uroepithelial cell lines differing in the degree of transformation were studied and compared by statistical evaluation of the harmonic amplitudes describing mathematically the cell shape: Two cell lines derived from human transitional cell carcinoma (TCC): 1. Hu 1703S classified as TGr I, 2. Hu 1703He classified as TGr III. It was found that these two cell lines differ in all harmonics. Two cell lines derived from morphological normal human bladder epithelium: 3. HCV-29 classified as TGr II.(ABSTRACT TRUNCATED AT 400 WORDS)

Carcinoma, Transitional Cell↗
PubMed1984

Crystallinity of mineral deposited in arterial walls in the course of arteriosclerosis in diabetics and in patients with normal carbohydrate metabolism.

A comparison was made of the amount and crystallinity of mineral deposited in the course of the arteriosclerosis in arterial walls in diabetic patients and in individuals with normal carbohydrate metabolism. The macroscopically unchanged tunica intima, fibro-lipidic plaques and bone-like lamellae were taken from aorta thoracalis , aorta abdominalis, arteriae femorales and arteriae coronariae in the course of the autopsy of 17 insulin dependent diabetic individuals and of 9 persons with arteriosclerosis, but with normal carbohydrate metabolism, called control group. The total inorganic constituents in the three kinds of samples were determined by ashing of dried samples at 600 degrees C for 6 h. Crystallinity of mineral was defined as the ratio of the spins connected with the radiation-induced stable paramagnetic centers present in the crystalline lattice of hydroxyapatite crystal, to the total ash content of the sample. An increase in the amount and in the crystallinity of deposited mineral was observed when consecutive stadia of development of arteriosclerotic lesions were compared. This phenomenon resembles the maturation of mineral observed in the course of bone development.

Aged↗
PubMed1983

Synthesis and secretion of proteoglycans by cultured chondrocytes. Effects of monensin, colchicine and beta-D-xyloside.

Chondrocytes, isolated from elastic ear cartilage of young rabbits, were grown in monolayer cultures in Ham's F-12 medium. Synthesis and secretion of macromolecules were monitored by labelling with radioactive precursors and the effect of monensin and other experimental agents was investigated. Monensin caused an inhibition of the incorporation of precursors into macromolecular material and a moderate intracellular accumulation when used in higher concentrations. The effect was more pronounced for 35SO4 than for 3H-labelled glucose or proline. p-Nitrophenyl-beta-D-xyloside alleviated this inhibition to some extent, but there was a concomitant increase in the amount of intracellular labelled material. Colchicine and monensin together caused a severe inhibition of the incorporation of 35SO4 and a marked shift of the label to the intracellular compartment. Colchicine also increased the sensitivity of the cells to monensin, lowering the minimal effective concentration about one order of magnitude. The latter results are consistent with the idea that cytoplasmic microtubules have a stabilizing function on the secretory pathways and, that their removal by colchicine, causing a 'randomizing' of the Golgi complex, makes these pathways more vulnerable to monensin.

Animals↗
PubMed1983

Application of optical diffractometry in studies of cell fine structure. Comparison of arterial smooth muscle cells in contractile and synthetic state.

Arterial smooth muscle cells in contractile and synthetic state were analyzed by optical diffractometry. Cell sections (80-90 nm) were photographed in an electron microscope and diffraction patterns of the plates (negatives) were produced using a helium-neon laser. Radial and angular distributions of light intensity in the diffractograms were measured and digitized using an electronic detector plate consisting of ring- and wedge-shaped photosensitive elements; radial distributions provide information about size of structures and distances between them and angular distributions about spatial orientation of structures in the images. Micrographs of nuclei and cytoplasm were analyzed separately (40-50 plates in each group). Computerized statistical analysis of radial distributions of light intensity showed that the nuclear chromatin pattern differed between cells in contractile and synthetic state. The probability that the observed difference could have arisen purely by chance was less than 10(-5). Computer-aided classification to the a priori known cell group was correct in 96.5% of the cases. Analysis of radial distributions of light intensity similarly showed marked differences in cytoplasmic structure between cells in contractile state (dominated by bundles of myofilaments) and synthetic state (dominated by cisternae of rough endoplasmic reticulum). The probability that the observed difference could have arisen purely by chance was less than 10(-5). Computer-aided classification to the a priori known cell group was correct in 92.0% of the cases. In contrast, analysis of angular distributions of light intensity did not indicate any statistically significant differences between contractile and synthetic state cells. A likely reason is that both myofilaments and cisternae of rough endoplasmic reticulum were arranged in parallel. The results demonstrate that optical diffractometry is a useful method for image analysis in studies of cell fine structure. It provides information about size and orientation of structures with poorly defined shape and is particularly well suited for studies on cell differentiation and effects of pharmacological and other experimental treatments on cell fine structure. It represents an alternative and a complement to stereology for quantitative and objective evaluation of morphological data.

Animals↗
PubMed1982

Optical diffraction as a tool for semiautomatic, quantitative analysis of tissue specimens.

Optical diffraction was tested on electron micrographs of normal and malformed myelin sheaths as a method for semiautomatic quantitative analysis of tissue specimens. Both normal and malformed myelin sheaths were chosen for the analysis because of their characteristic internal structure and its alteration as a result of malformation. Optical diffraction patterns were obtained by means of an optical diffractometer coupled with a digital detector. The spacing and arrangement of the components of various types of myelin sheath were automatically calculated and determined and the results were verified with discriminant analysis. Out of 27 parameters of the radial and out of 25 parameters of the angular distributions of diffracted light intensity, 6 and 11, respectively, were found to have good discriminative power and were used for classification of myelin sheaths. The accuracy of automatic classification was tested by comparison with myelin sheath types of known origin. The samples visually similar by their appearance, e.g. control and regenerating myelin sheaths, were automatically classified with accuracy of 69%, whereas others were classified appropriately with 88-100% accuracy. It is believed that this kind of analysis may successfully be applied for specimens of other tissues and/or organs.

Animals↗
PubMed1982

Application of the optical Fourier transform for analysis of the spatial distribution of collagen fibers in normal and osteopetrotic bone tissue.

Optical Fourier analysis was applied for evaluation of the differences between normal and pathologically changed bone tissue. Collagen fibers were used as markers of bone structure. To prove the usefulness of this technique for objective mathematical analysis of these differences the spatial spatial distribution of collagen fiber bundles was evaluated in normal and osteopetrotic bone. The variation in the spatial distribution of collagen fiber bundles in cross sections of femur diaphyses was evaluated quantitatively by optical diffraction three groups of Fatty Orl-op strain rats, i.e. phenotypically normal animals, osteopetrotic (op/op) mutants and op/op-mutants cured by transplantation of normal syngenic bone marrow. The histological sections of decalcified bone were stained with Sirus-Red and then photographed under polarizing microscope. The Sirus-Red staining was used to enhance the natural birefringency of collagen fibers. Diffractograms obtained from microphotographs of selected bone section areas, i.e. outer and inner circumferential lamellae and haversian bone of normal and cured op/op animals as well as whole cortical bone and woven bone filling the medullary cavities in op/op mutants were analysed separately. Diffractograms contain summarized information on the size and relative position of these structures in histological sections. The radial and angular distribution of light energy were evaluated for each diffractogram with an electronic detector. The obtained distributions were described by several sets of parameters concerning the position, level and shape of local maxima and minima. Out of these parameters five with the highest discriminant power were chosen for further mathematical analysis. This analysis was based on the calculation of the position of centroids in the multidimensional space described by the linear functions of the chosen parameters for each of the evaluated bone section areas. The centroids (mean values of discriminant scores of each group) represent the centers of gravity of the analysed groups, while the separation of the centroids tested by the F-test illustrates the differences between the respective groups of selected bone section areas. A high level of separation of centroids was found when osteopetreotic bone was compared with normal one, what means that the spatial distribution, size and interstructural distances between the collagen fiber bundles in bone tissue in these two groups of animals differ markedly. A similar situation was observed when osteopetrotic bone was compared with bone tissue obtained from op/op mutants cured by transplantation of normal syngenic bone marrow. On the other hand, the level of separation of centroids was low when bone tissue of cured op/op mutants was compared with the control one, a finding which corresponds to the less pronounced histological differences between these two groups of animals. Computer-aided classification on single microphotographs of selected bone section areas to the known a priori type of bone tissue was performed...

Animals↗
PubMed1982

Polarizing microscopy of Picrosirius stained bone sections as a method for analysis of spatial distribution of collagen fibers by optical diffractometry.

Cross sections of femur diaphysis obtained from control and osteopetrotic rats were stained with hematoxylin-eosin (HE) and Picrosirius (SR). Analogous selected areas of bone sections photographed under a polarizing microscope were analysed by optical diffractometry. Since the collagen fibers are a good marker for the structure of bone tissue, their spatial distribution evaluated by optical diffractometry provides information on the tissue architecture. The Picrosirius staining technique enhances the natural birefringency of collagen fibers. Therefore, in the polarizing microscope, pictures of high contrast are obtained. This procedure, by increasing the amount of information in the image, increases the quantity of the data obtained by optical diffractometry in comparison with the HE staining method. The results obtained prove that SR staining combined with polarizing microscopy might be useful for optical diffractometry in analysis of the spatial distribution of collagen fibers in all connective tissues, where they could serve as markers of tissue architecture.

Animals↗
PubMed1981

Crystallinity of tissue mineral as evaluated by electron spin resonance spectometry.

Crystallinity of bone mineral as well as mineral deposited in the course of pathological calcification was measured by the use of electron spin resonance (esr) spectometry in tissue samples previously irradiated with ionizing radiation. Defects caused by irradiation in the crystalline lattice of hydroxypatite crystals are paramagnetic in nature, stable and connected with the crystalline fraction of tissue mineral only. The crystallinity coefficient, defined as the ratio of these paramagnetic centers to the total ash content, changes in bone in the course of development, ageing and in pathology. The accuracy and sensitivity of esr method developed for estimation of crystallinity of tissue mineral are discussed. Examples are given for application of the esr method for the research of normal and pathologically changed mineralizing tissues. The sensitivity of the esr method permitted the measurements of crystallinity of single isolated osteons in different stages of calcification, as well as the evaluation of crystallinity of mineral deposits in subcellular fractions, e.g., calcium-loaded mitochondria. The developed technique might be applied in clinical research for diagnosis and monitoring of therapy in selected bone diseases.

Aging↗
PubMed1981

[Diagnosis of cysticercosis using the intradermal test].

56 healthy rabbits were inoculated with the active Cysticercus each into lateral ventricle of the brain, the white substance of the brain and into Spatium subarachnoideale. Rabbits were killed after 32 weeks p.i. Then their brains were macro- ev. microscopically investigated. The positive seroreactions (ringpreciptitation reaction and the latex-test) were observed at the beginning of the second week after infection. Curve of the titers was revealed two tops: first one between 6-7 weeks, second one 16-22 weeks after infection. All rabbits except one reacted positive on the application of the Cysticercus antigen. The control group consisted of the 8 rabbits revealed negative serological and immunological reactions. Three groups of the cerebral cysticercosis, dependent on the localization of the cysticerci in the brains, were observed: oedema form, hydrocephalous form and tumor form. These forms as well as the symptoms are similar to the human cerebral cysticercosis.

Animals↗
PubMed1980

Crystallinity of human pineal calcospherulites.

Crystallinity of mineral in human pineal calcospherulites was determined by electron spin resonance spectrometry after irradiation of the samples with gamma rays in a 60Co-source. The radiation-induced stable paramagnetic centers in the crystalline lattice of hydroxyapatite crystals were used as a marker of the crystalline fraction and related to the total mineral content. The crystallinity of pineal sand is higher than that of compact bone. The numerical value of the crystallinity coefficient depends on both the average crystal size of hydroxyapatite and the percentage of the crystalline fraction in the total amount of mineral. Literature data show that the average size of hydroxyapatite crystals in pineal sand are smaller than in bone tissue. It is, therefore, concluded that the higher crystallinity of pineal acervuli is due to the lower percentage of the submicrocrystalline fraction in their mineral.

Aged↗
PubMed1980

Heterotopically induced osteogenesis in osteopetrotic rat mutants. A preliminary report.

Heterotopic osteogenesis was induced in three groups of Fatty Orl-op strain of rats, i.e., osteopetrotic (op/op) mutants, op/op mutants cured by previous transplants of normal bone marrow, and phenotypically normal littermates. Allogenic decalcified bone matrix was used as inductor. The osteopetrotic mutants from heterotopic ossicles containing bone and bone marrow under the influence of inductor. After four weeks the signs of remodelling processes of induced bone tissue are observed: osteoclasts are found in the Howship lacunae, and bone marrow is formed in the spaces adjacent to the bone trabeculae. The induced bone marrow contains megakaryocytes, cells of the erythropoietic and granulopoietic lines as well as some infiltrating lymphocytes. The important finding is that the amount and the histological structure of bone tissue and bone marrow induced heterotopically in the osteopetrotic animals were similar to that observed in both control groups. It is concluded that 1) osteopetrotic mutants have the same potency to form heterotopic bone tissue under the influence of inductor as the control animals, and 2) remodelling processes in four-week-old, induced ossicles are normal and differ in this respect from defective rebuilding of the orthotopic bones of op/op hosts.

Animals↗