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Platelet pathology in patients with idiopathic scoliosis: Ultrastructural morphometry, agrregations, x-ray spectrometry, and biochemical analysis.

The fundamental similarity between platelets and muscle, suggested the possiblity of a shared defect in idiopathic scoliosis, a genetic disease with lateral deformity of the spine in which there is an elevation of calicum concentration in muscles and platelets. A variety of platelet tests revealed the following abnormalities: (1) Electron microscopic x-ray analysis and x-ray fluorescence spectrometry showed a 2- to 3-fold increase in calcium and phosphorus in whole cells and in individual dense bodies. (2) Electron microscopy morphometry revealed an increase in electron-opaque bodies in air-dried cells; granules and microtubules were unchanged. There were more large cells and membranous complexes. (3) Aggregations with epinephrine and ADP were depressed in some patients. (4) Proteins (total and contractile) and myosin. ATPase activity in centrifuged fractions of platelets were decreased in the cytosol and increased in the fraction containing membranes and granules. The correlated findings suggest that platelets in idiopathic scoliosis have a mild calcium transport defect related to membrane and/or contractile protein metabolism. This investigation also shows that platelets may be used to advantage in diagnosis and research of muscle diseases.

Actins↗

[Multivariable EDV-program for light and electron microscopic morphometry].

A computer program for morphometry and stereology in which conventional morphometric and stereological data acquisition (direct linear measurement, point counting technique) is used and which is designed for variable application of conventional and electron microscopy is presented. There are several program variants and magnification stages each designed for a different structural level. There is fixed number of essential subcellular, cellular and extracellular structures for different kinds of tissue (myocardial, liver and connective tissue), but this number can be enlarged with each stage of magnification and for each measured parameter by reserving free places for evaluating additional structures. Thanks to the extended program and to the fact that these free places are interchangeable, the program can be used independently of the kind of tissue examined and of the kind of morphological information sought. The program structure is realized by means of a master control and 4 external subprograms. The statistical analysis of the morphometric stereological parameters is calculated by image, case by case or group by group. The mean values of the calculated primary and secondary parameters, the standard errors and the standard error/mean value ratio are printed out. All common stereological parameters and the essential cellular and extracellular structures of different kinds of tissue are described in quantitative terms with the aid of the program.

Computers↗

[Possibility of the morphometry of the neuronal body and nucleus in cryostat sections of nerve tissue].

A method of fixation by the Carnoy liquid and of gallocyanine staining is proposed in order to obtain clear-cut boundaries of the neuron body, nucleus and nucleolus in cryostat sections of non-fixed frozen nerve tissue, for subsequent morphometry of the body and nucleus of nerve cells. In the resulting sections, the clearness of boundaries of the body and nucleus in a neuron is sufficient for measuring its diameter with the coefficient of variation not exceeding 2.5%.

Animals↗

Image processing in pathology. XII. Automated morphometry of tumour cell nuclei in hypernephroma: comparison between renal tumour and late metastases.

Changes of nineteen features of tumour cell nuclei in hypernephroma and three following late metastases were investigated by use of the automated image system. Histograms of nuclei features from primary tumours and metastases were compared. They showed objectively an increased cellular dedifferentiation during metastases. Various aspects of the capability of automated morphometry for investigation of tumours are discussed.

Adenocarcinoma↗

"Arabidobrassica": Evidence for intergeneric somatic hybrid nature from electron microscopic morphometry of chromatin.

The new method of species and hybrid characterization and determination by quantitative morphometry of chromatin texture in ultra-thin sections of nuclei was applied to the somatic hybrid "Arabidobrassica" and its parents, Arabidopsis thaliana and Brassica campestris. The somatic hybrid exhibits the average amount of condensed chromatin, if compared to the proportion of condensed chromatin in nuclei of the parent species. This result corresponds to similar findings in natural allopolyploids and is consistent with the assumption that "Arabidobrassica" is a true intergeneric hybrid.

Brassica↗

Ultrastructural morphometry of the secretory granules in thyroid medullary carcinoma.

The authors studied the ultrastructural morphometry of secretory granules in four cases of thyroid medullary carcinoma. An inhomogeneous granules population was found with values ranging from 70 to 450 nm and mean values between 171.4 +/- 31.8 and 222.7 +/- 28.1 nm, respectively. Plurimodal frequency distribution curves showed peaks at 80, 90, 130, 160, 200, 240 and 250 nm. Two possibilities explaining the variation in granules size are discussed: the size of granules could depend on the maturation degree and functional activity or, more probably, there are several kinds of granules containing various biogenic substances, such as calcitonin and ACTH. The granules might play a role in the genesis of amyloid fibrils as well, some granules being located around amyloid fibrils.

Carcinoma↗

Computed tomographic morphometry of the lumbosacral spine of dogs.

In a 5-year prospective study, computed tomographic (CT) morphometry of the lumbosacral vertebral canal was performed on 42 large-breed dogs (21 controls and 21 dogs with lumbosacral stenosis). Dogs were allotted to 4 groups. Group 1 (n = 13) consisted of cadaver specimens obtained from dogs that died or were euthanatized for reasons unrelated to the spine; group 2 (n = 8) consisted of live dogs with no history of clinical signs related to the spine and with normal neurologic examination findings; group 3 (n = 10) consisted of dogs with surgically confirmed lumbosacral stenosis; and group 4 (n = 11) consisted of dogs with suspected lumbosacral stenosis that were managed conservatively. The CT scans were performed, using 5-mm contiguous slices obtained perpendicular to the vertebral canal, from the midbody of the 5th lumbar vertebra through the caudal endplate of the sacrum (L5-S3). Lumbosacral lordosis was minimized in all dogs by positioning them in dorsal recumbency with the hind limbs flexed. A tuberculin syringe calibration phantom was placed within the scanning field of view, parallel to the axis of the spine. In each dog, 11 CT slice locations within the lumbosacral spine were evaluated. At each slice location, sagittal plane diameter, dorsal plane diameter, and transverse area of the vertebral canal, vertebral body, and calibration phantom were measured, using the CT computer's software programs for distance and area calculation. Window/level settings were constant, and all measurements were made by the same operator (JCJ). Accuracy of calibration phantom CT measurements was 100% for sagittal and dorsal plane diameter and was 85% for transverse area.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Quantitative nuclear morphometry, Markovian texture descriptors, and DNA content captured on a CAS-200 Image analysis system, combined with PCNA and HER-2/neu immunohistochemistry for prediction of prostate cancer progression.

One hundred and twenty-four localized prostate cancer patients operated on at Johns Hopkins Hospital (JHH) since 1975 were identified. The sample was optimized for evaluation of prostate cancer progression. Based upon accurate clinical histories, these radical prostatectomy patients included 50 progressors and 74 non-progressors using appearance of serum PSA as an indication of recurrence (mean follow-up = 8.6 +/- 1.8 years, range 7-15 years). All patients included in the study had no involvement of their seminal vesicles or lymph nodes at the time of prostatectomy. Average time to progression was 3.6 +/- 2 years, range of 1-8 years. Using paraffin-embedded specimens, several five micron sections were cut and placed on Probe-On slides; one slide was H&E-stained and the other was Feulgen-stained. The H&E and Feulgen-stained slides were screened and "dotted" by pathologists at JHH and CytoDynostics, Inc. A CAS-200 Image analysis system (Cell Image Systems, Elmhurst, IL) equipped with a Cell Measurement Program version 1.2 beta, was used to capture the Feulgen-stained images and to perform the calculations. From the "dotted" areas, 150 cancer cells were selected for measurement of DNA content and 27 nuclear morphometric shape and size factors, including 21 Markovian chromatin texture variables. Additional sections were used for immunochemistry staining with an alkaline phosphatase streptavidin-biotin complex stain to detect and quantitate cancer cells binding monoclonal antibodies directed against proliferating cell nuclear antigen (PCNA) and HER-2/neu antigen. All data were entered into a statistical program (STATA) for further analysis and univariate and multivariate statistical analysis was performed using logistic regression and its stepwise variant. The biomarkers of greatest utility to detect progressors when analyzed univariately included post-operative Gleason score (p = < 0.0001), HER-2/neu antigenicity (p = 0.0147), CAS-200 DNA ploidy (p = 0.008), and twelve Markovian nuclear texture and shape features (p = < 0.0001), whereas PCNA (p = 0.160) failed. The optimal set of nuclear morphometry progression tumor features were selected using backward stepwise logistic regression estimate analysis which drops variables due to collinearity. Although post-operative Gleason score is a strong univariate predictor of progression, DNA ploidy and HER-2/neu contributed significantly to further stratification of higher risk groups within the low Gleason score subpopulation. The best Markovian features combined with post-operative Gleason score generated sensitivity = 90%, specificity = 96%, positive predictive value = 94%, negative predictive value = 93% and the area under the receiver operator curve was 0.975.

Biomarkers, Tumor↗

Automated sperm morphometry analysis (ASMA) in the rabbit.

New methods of specimen preparation were developed and a new method of objective, automated sperm morphometry analysis (ASMA) was performed to reduce the technical variation in the rabbit sperm morphology assay. The optimal staining procedure was a modified GZIN stain, which allowed the ASMA instrument to accurately recognize the distal end of the sperm head and to achieve the highest sperm recognition rate (94%). Washing and resuspending sperm to a standard concentration increased the number of sperm per microscopic field in low-concentration samples and reduced the field-to-field variation in all samples. Washing also decreased the number of sperm recognition errors by the ASMA instrument. Mean metric measurements for all sperm were: length, 7.38 microns; width, 3.91 microns; width/length, 0.53; area, 22.10 microns; and perimeter, 19.20 microns. Within-specimen coefficients of variation (CVs) ranged from 0.8% to 5.5% and between-animal CVs ranged from 2.7% to 7.5%. The use of standard specimen preparation techniques and ASMA technology can significantly reduce the technical variation in the rabbit sperm morphology assay.

Animals↗

[Multiparametric cytometric analysis of hepatocellular carcinoma and its allied lesions combining DNA ploidy analysis with morphometry using DAPI/HP double staining].

Hepatocellular carcinomas (HCC) of various differentiation, Edmondson (Edm) grade I-IV, often based upon viral infectious hepatitis transformed through regeneration, nodular and/or dysplastic adenomatous hyperplasia. Aspiration cytology, even needle biopsy for such lesions, is very difficult to interpret. We have been studying multiparametric cytologic analysis using DAPI/Hematoporphyrin (HP) staining for the cece samples provided from exactly trimmed 50-microns thick paraffin sections, employing a new software program which we developed, combining DNA ploidy and cell protein content analysis with morphometry. DAPI indicates DNA content and nuclear size and HP indicates cell protein content and cell size. From 21 cases of HCC, 43 nodules or masses including multiple lesions and nodule in nodule were analyzed. Surrounding non-neoplastic hepatic tissues were examined as the control specimens. On DNA histogram, cases with the peak over 2.4 c are identified as aneuploid (Aneupl). Cases of Edm I, or II, 6 of 8 samples showed Aneupl., all of 11 cases of Edm II, 11 cases of Edm III, and one of Edm IV showed Aneupl. Multinodular foci in the same cases and also nodule in nodule often showed different modes of Aneupl. This suggested heterogenetic carcinogeneity. Noncarcinomatous foci often showed Aneupl. modes, which may indicate malignant transformation or malignant potential.

Carcinoma, Hepatocellular↗

Morphometry of experimental lung contusion: an improved quantitative method.

Alterations of lung tissue following pulmonary contusion are mainly assessed qualitatively. Quantitative examinations were limited up to now, to investigations in X-ray and computer tomography images, since morphometry of contused and normal biotic lung tissues seemed to be methodologically unfeasible. In order to overcome this shortcoming, the lungs of pigs were examined after unilateral lung contusion. In semithin sections, the thickness distribution of the interstitium and the volume distribution of the alveolar cavities were studied. Using an image analysis system a sequence of detection, erosion and skeletonisation steps were used to recognize the septal tissue correctly. The percent distributions were calculated from chord measurements after a sequence of erosion steps. It turned out that this technically simple procedure enabled highly reliable and valid results to be obtained, concerning the morphometric analysis of the alterations in the contused, as well as in the non-injured lungs.

Animals↗

Nuclear morphometry in differential diagnosis of renal oncocytoma and renal cell carcinoma.

PURPOSE: Morphometric distinction of renal oncocytoma and renal cell carcinoma was attempted. MATERIALS AND METHODS: Nuclear morphometric measurements were done from sections of 16 histologically diagnosed renal oncocytomas and renal cell carcinoma controls (grade 1 or 2, stage T1 to 2N0M0). RESULTS: Oncocytomas could be distinguished from carcinomas on the basis of nuclear morphometry in more than 80% of the cases. Especially, the shape descriptors (form factors) were significantly different between oncocytomas and carcinomas. CONCLUSIONS: Morphometrically determined nuclear shape descriptors can be used to support diagnostic decisions in problematic cases.

Adenoma, Oxyphilic↗

[The value of nuclear morphometry in analysing heterogeneity of grade-II colorectal adenocarcinomas].

A nuclear morphometric analysis was performed on the histologic sections of 39 grade-II, 17 grade-I and 8 grade-II colorectal adenocarcinomas. The parameters selected were nuclear area, nuclear perimeter, nuclear volume and standard deviation of nuclear area. The results showed that grade-II colorectal adenocarcinomas were a group of heterogeneous tumours and they could be divided into various subgroups by nuclear morphometry so that their biological behaviour could be better reflected.

Adenocarcinoma↗

Comparison of soft tissue facial morphometry in children with Class I and Class II occlusions.

Three-dimensional soft tissue facial morphometry was investigated in a sample of 167 children aged 6 to 9 years by using a new noninvasive computerized method. For each child, 16 cutaneous facial landmarks were automatically collected by a system consisting of two infrared CCD cameras, real-time hardware for the recognition of markers, and software for the three-dimensional reconstruction of the x, y, and z coordinates of landmarks. From these landmarks, 15 linear and 10 angular measurements and five linear distance ratios were computed. For each age class, mean values were computed for all children with a bilateral Angle Class I occlusion (modified according to Katz) and compared with values obtained in children with a bilateral Class II occlusion. Most of the differences involved three-dimensional angular measurements: Class II children had more convex faces in the sagittal plane and a less prominent mandible than did Class I children. No differences were found in the linear measurements. Only the lower facial height ratio was different between the two occlusion groups, but the difference was not consistent among all the age groups.

Cephalometry↗

Combined strategy of conventional cytogenetics, fluorescent in situ hybridization and chromosome morphometry for analysis of parotid gland tumor.

The limiting factors in conventional cytogenetic analysis of cell culture, especially of solid tumors, include insufficient metaphases, overgrowth of abnormal mitotic cells by normal cells, and suboptimal quality of harvesting and banding. Despite the availability of numerous protocols to induce G-banding, as well as Q-, R-, and C-banding, occasions still arise in which the analysis is severely limited by these factors and incomplete conclusions are often drawn as to the precise nature of the chromosomal abnormality, if indeed any can be detected. By adopting a rational approach of (1) close monitoring of cultures and rapid harvesting as soon as it is feasible, and (2) analysis of available metaphases by a combination of the GTG technique, fluorescent in situ hybridization (FISH), and chromosome morphometry using a graphic arts tool, a significant improvement in success rate may be more readily achieved. Here pathological and cytogenetic data are presented of a case of parotid gland carcinoma ex mixed tumor with the karyotype of 46, XX, del(5)(q12), dir ins(8;5)(q12;q12qter), add(12)(p13)/46, XX. This case is utilized to illustrate the importance of application of our combined strategy.

Chromosomes, Human, Pair 12↗

[Usefulness of nuclear morphometry as predictive factor of progression in bladder papillary carcinoma].

Lately, nuclear morphometry methods are being used to analyze the malignant potential of tumoral cells. This paper presents a retrospective study of a series of 163 patients with the three histological grades (WHO), of superficial papillary transitional cell carcinoma (pTa-pT1) of the bladder with a mean follow-up of 60 months. A morphometric analysis of the initial tumour biopsies was made. Mean nuclear area (NA) and its standard deviation (NASD) were measured in a semi-automatic image analyzer (MOP-videoplan, Kontron). NA and NASD show increasingly higher values with increased grade (p < or = 0.0001). NA and NASD values raise in those progressing to muscle-infiltrant stage; tumours were, then, divided into two morphometric groups: small NA (< 50 microns 2) and large NA (> 50 microns 2); the difference between them being statistically significant. The likelihood of no tumoral progression at 30, 60 and 90 months was, respectively, 96, 94 and 92 percent, form small NAs, and 76, 65 and 64 percent for large NA (p < or = 0.0001).

Carcinoma, Transitional Cell↗

The site, severity, and distribution of bronchiolar obstruction in lungs with chronic obstructive pulmonary disease. Morphometry and computer-assisted three-dimensional reconstruction of airways.

Autopsy lungs from 10 patients with various types of chronic obstructive pulmonary disease were submitted to morphometry and computer-aided three-dimensional reconstruction of airways to study, by the former technique, the severity and the site of obstruction in the bronchioles and to visualize, by the latter, how far the whole airway bed was involved. The site of obstruction in the bronchioles was shown to be different between diffuse panbronchiolitis and bronchobronchiolitis obliterans. There was also a case where the entire bronchiolar tree was mildly but uniformly stenosed with mucous hypersecretion, chronic inflammation, and fibrosis, a pattern considered to correspond to small airways disease. Thus, the combination of the above techniques proved to be useful in studying the state of the airway bed in chronic obstructive pulmonary disease, a task not accessible by the usual microscopic examination of a single lung section.

Adult↗

[The computed tomographic morphometry of the brain in minimal brain dysfunction in children 5 to 7 years old].

Computed tomography (CT) morphometry of the brain in preschool children in the critical period reflects the dynamic process of the brain formation, multidirectional laterality of the hemispheres. Close relationships between neuropsychological and CT results demonstrate general tendency of the neuro-ontogenetic process comprising reciprocal interaction of functional activity and structural arrangement of the brain compartments.

Aging↗