Predictive value of discriminant analysis of monocyte ultrastructure in malignant lymphoma

Discriminant analysis was applied to morphometric data obtained from ultrastructural studies of blood monocytes from 20 normal subjects, 23 patients with Hodgkin's disease and 12 patients with non-Hodgkin's lymphoma. The aim was to assess the efficiency of predicting subject groups from such data. The analysis, performed on a microcomputer using a standard statistical package, considered nuclear volume, nuclear surface area, nucleolar volume, nucleolar surface area, nucleolar volume fraction, number of nucleoli per section, cell surface area, mitochondrial surface area and subject age. The overall agreement between predicted and actual subject groups was 64%; considering only normality and disease, the agreement was 80%. While the predictive value of such data from circulating monocytes would appear insufficient for diagnostic purposes, discriminant analysis as used here might be of value in indicating the state of host defense in malignancy.     

A new method of a morphometric study of the kidney using a computer-assisted morphometric complex "Diamorph"]

The use of a computer--based morphometric complex "DIAMORPH" for automated morphometric analysis of renal glomeruli is described. By combining the principles of morphological and functional approaches to diagnosis with the capabilities of present--day research equipment, a new informative morphometric parameter--the extent of capillary--mesangium contact--has been identified. With this parameter and its derivatives, it has been possible to differentiate, in a statistically significant way (p less than 0.01), healthy subjects (controls) from patients with manifest diabetes mellitus of up to I year in duration who could not be distinguished through light microscopic or conventional morphometric studies.     

Morphological,cytochemical, functional,and proliferative characteristics of four murine macrophage-like cell lines

The aim of the present study was to obtain objective data on the morphology and quantitative information about other characteristics of murine macrophage-like cell lines J774.1, PU5-1.8, WEHI-3, and P388-D1, and to compare the findings with those in resident and exudate macrophages collected directly from mice. Fetal fibroblasts were included to serve as controls.Evaluation of the morphological data showed that the cell lines J774.1 and WEHI-3 are almost identical in most respects, that the cells of P388-D1 differ widely from both of the former lines, and that the morphometric parameters of cell line PU5-1.8 occupy an intermediate position. The cells of the P388-D1 line show the most similarity to resident and exudate macrophages, and cell lines J774.1 and WEHI-3 the least. Fetal fibroblasts had divergent values for all morphometric parameters. Good correspondence was found when the quantitative data obtained by morphometric analysis of the cells in question were compared with the morphological pictures.No gross differences as to cytochemical characteristics were found between the cells of the four cell lines, except for 5′-nucleotidase activity. The occurrence of IgG receptors and the ingestion of EIgG were also similar, but the percentage of cells with C3b receptors was much lower in two of the cell lines (WEHI-3 and P388-D1) and the level of EIgMC ingestion was very much higher in one (J774.1) compared with both the other cell lines and the resident and exudate macrophages. The ingestion of opsonized bacteria and latex varied widely within and between the cell lines. Quantitative data on the binding of monoclonal antibodies by the cells of the macrophage cell lines and the resident and exudate macrophages showed a wide variation. The doubling time of the cell lines is on average 1 day; distinct differences were found between these lines with respect to the lag-time of proliferation after replating.Cluster analysis and statistical analysis of morphological and other characteristics gave insight into the degree of resemblance between the cells of the four cell lines on the one hand and the resident and exudate macrophages on the other.     

International study on Artemia. LIV. Morphological study of Artemia with emphasis to Old World strains. II. Parthenogenetic populations

Eleven morphometric and one meristic character in 15 parthenogeneticArtemia populations have been studied by using discriminant andcluster analysis as well as scanning electron microscopy.Discriminant analysis revealed five main groups of morphologicalpatterns: (i) the coastal Chinese populations together with apopulation from Kazakhstan, (ii) the inland Chinese salt lakepopulations, (iii) the Greek populations, (iv) one African populationfrom Namibia and (v) a Chinese population from Xuyu (Jiangsuprovince). Cluster analysis was not always in agreement withdiscriminant analysis and these results are discussed.     

A morphometric analysis of cytological features of tall cell variant and classical papillary carcinoma of the thyroid

In order to assess whether morphometric parameters could be of value in distinguishing between tall cell variant and classical pattern of thyroid papillary carcinoma, the fine needle aspiration cytology (FNAC) samples of 14 cases were analysed using Arcimage 5 software on an Acorn computer. Histological examination of the specimens allowe classification of nine of them as classical pattern and the remaining five as tall cell variants. The nuclear diameter (NDD) and standard deviation distribution (NDSDD), th nuclear area (NAD) and standard deviation distribution (NASDD), and the nuclear/cytoplasmic ratio (NCR) were assessed on May-Grunwald-Giemsa stained smears. Statistical analysis was performed by use of one-way analysis of variance (ANOVA) of the two groups as identified by histology. Whilst NDD (P = 0.007), NAD (P = 0.015) and NADSD (P = 0.026) all appeared statistically significant, NDSD (P = 0.06) and NCR (P = 0.71) were not. The cytological diagnosis of papillary carcinoma is established and reproducible, but morphometric data on the thyroid have so far focused on the differential diagnosis between benign and malignant nodules. The choice of simple morphometric parameters appears to be helpful in the preoperative distinction between the classical pattern and tall cell variant of papillary carcinoma.     

Patterns of morphological and genetic variability in UK populations of the shore crab, Carcinus maenas Linnaeus, 1758 (Crustacea: Decapoda: Brachyura)

Previous research has identified extensive inter-population variability in the morphology of the shore crab (Carcinus maenas L.). To determine the source of this variation (genetic or environmental), morphological and genetic data were analysed from crabs collected from eight sites around the coast of the UK. Ten morphometric traits were measured from over 800 crabs and the degree of morphological similarity among sites was calculated using multivariate techniques. Allozyme electrophoresis was used to investigate patterns of genetic similarity. Extensive morphological variability was detected: eight out of the ten morphometric traits analysed were useful when discriminating between crabs from each site. Discriminant function analysis revealed that over 35% of individuals could be classified to their site of origin on the basis of their morphology. In contrast, the allozyme analysis revealed low levels of genetic variability, both within the meta-population and among the crab population at each site. Pairwise comparisons revealed a moderate correlation between the degree of morphological and genetic similarity of crabs at each site, which suggests that the observed phenotypic variability has a genetic component. However, only around 20% of the phenotypic variability detected was associated with the patterns of genetic similarity. This means that patterns of morphological variability in this species are largely determined by the local environmental conditions: local factors could have a within-generation selective influence on mean trait values or C. maenas may exhibit phenotypic plasticity.     

Morphometric study of two species of wood miceApodemus sylvaticus andA. flavicollis (Rodentia: Muridae): traditional and geometric morphometric approach

Skull shape and size were investigated in 311 specimens ofApodemus sylvaticus (Linnaeus, 1758) andA. flavicollis (Melchior, 1834), using traditional and geometric morphometric approach. We searched for potential morphometric diagnostic criteria between these morphologically similar mice and we an alyzed the interpopulation morphometric variation. Specific status of all individuals was identified by scoring four diagnostic allozyme markers. Discriminant analysis based on 16 cranial measure ments as well as that based on land mark data for ventral side of the skull discriminated in dividuals with 99.7% accuracy. The classification functions useful for practical discrimination of unknown specimens are given. Our data showed neither clear intraspecific geographical pattern among studied populations nor evidence for character displacement in syntopic populations ofA. sylvaticus andA. flavicollis.     

Morphometry in evaluating large cell malignant lymphomas in lymph node biopsies

The values of some morphometric cell parameters were analysed in 12 cases of large cell non-Hodgkin's lymphoma (centroblastic and immunoblastic lymphoma and histiocytic sarcomas) in order to test the utility of the morphometric method as an additional element to the histologic diagnosis and to its further accuracy. The nuclear and nucleolar area, the number of nucleoli per nuclear cross section and the nucleus to nucleolus ratio were investigated. The morphometric data were obtained on histograms using the Giemsa stained sections. Mean values and standard deviations were calculated and statistically analyzed. Statistical evaluation of data showed significant differences in the three groups. Our results proved the usefulness of the morphometric method as a helpful technique of differentiating the large cell malignant lymphomas.     

Geographic patterns of genetic and morphological divergence amongst populations of a river crab (Decapoda, Potamonautidae) with the description of a new species from mountain streams in the Western Cape, South Africa

Recent systematic research has revealed that Potamonautes brincki comprises two genetically and morphologically distinct population groups. The systematic affinities between these population groups have remained uncertain. In the present study, the relationship between the population groups was examined. Eleven populations were collected from high mountain streams in the Western Cape, South Africa and used in the genetic and morphological analyses. Allozyme electrophoresis of 13 protein coding loci separated two main population groups: group A (Cape Peninsular groups) and group B (Hottentot's Holland) at I  = 0.73. Two additional genetic groups were evident, with group B being conspecific to group A, and group D being conspecific to group C. Morphological examination of pleopod 1 and the terminal segment of the mandibular palp showed considerable differences between the two main population groups, with groups A and B being similar and groups C and D being similar. The morphometric data for the four main groups were examined using discriminant functions analysis and the two main groups were compared using analyses of covariance. Discriminant functions analysis showed a moderate degree of overlap between the groups. Additional morphometric data showed a clear discrimination between the two main population groups. The genetic and morphometric data sets exhibited congruent patterns of variation and the data showed the presence of a species boundary. A new freshwater crab species, P. parvicorpus sp. n., is described. The results of the present study are discussed in the light of historical and contemporary factors that are likely to have contributed to speciation.     

Is formalin fixation and ethanol preservation able to influence in geometric morphometric analysis? Fishes as a case study

Geometric morphometric analysis has increased in the recent years, turning into a powerful tool to explore shape and size variation. Several biological studies use specimens that have been through some kind of preservation, mainly formalin preservation, commonly used in biological collections. This study analyzed the effect of preservation in shape on two fish species: Eucinostomus argenteus and Pomadasys corvinaerformis. Twenty-nine individuals of E. argenteus and twenty-five of P. corvinaeformis were collected, photographed twice, preserved in 10 % formalin for 1 week, and then transferred to 70 % ethanol for 83 days. We evaluated three levels of error: (1) error of landmark digitalization, (2) error of taking the picture and storage in JPEG format, and (3) the formalin and ethanol fixation error using Procrustes ANOVA, Discriminant Analysis, and Principal Component Analysis. Significant difference between treatments was observed on both species with Procrustes ANOVA and Discriminant Analysis. In addition, Principal Component Analysis showed a separation between groups of treatment on both species. These results represent the first evidence of preservation effects in studies of geometric morphometrics and show that according to the statistical test utilized, the fixation could affect the shape variations in different ways and could lead the researcher to false results or wrong conclusions. Other methods to explore the shape variation of organisms previously fixed should be tested in order to assess their influence in geometric mophrometric studies.     

Is there any potential link among caspase-8, p-p38 MAPK and bcl-2 in clear cell renal cell carcinomas? A comparative immunohistochemical analysis with clinical connotations

Background

Management of endometrial precancerous lesions has been of much debate due to inconsistencies in their classification, natural history and histologic diagnosis. Endometrial hyperplasia constitutes a wide range of histomorphologic features associated with high intra and interobserver diagnostic variability. Although traditional microscopic diagnosis is by far the most applicable method and the gold standard for histomorphologic diagnosis, digitized image analysis has been used as a powerful adjunct to maximize the histologic data retrieval and to add some detailed objective criteria for correct diagnosis in difficult cases.

Methods

A series of 100 endometrial curettage specimens with diagnosis of endometrial hyperplasia or well differentiated adenocarcinoma were blindly reviewed by 5 pathologists; their intra and interobserver reproducibility determined and further compared to the objective morphometric data i.e. D-score and volume percent of stroma (VPS).

Results

The results were assessed using the weighted kappa statistics. Mean intraobserver kappa value was 0.8690 (99.44% agreement). Mean interobserver kappa values by diagnostic category were: simple hyperplasia without atypia: 0.7441; complex hyperplasia without atypia: 0.3379; atypical hyperplasia: 0.3473, and well-differentiated endometrioid carcinoma: 0.6428; with a kappa value of 0.5372 for all cases combined. Interobserver agreement was in substantial rate for simple hyperplasia (SH) and well differentiated adenocarcinoma (WDA) but was in fair limit for complex hyperplasia (CH) and atypical hyperplasia (AH). Intraobserver agreement was almost perfect. The specimens were divided in two groups according to the computerized morphometric analysis: Endometrial Hyperplasia (EH) ( D Score ≥ 1 or VPS ≥ 55%) and Endometrial Intraepithelial Neoplasia (EIN) (D-Score < 1 or VPS < 55%). Morphometric findings were closely compatible with routine WHO classification made by one expert pathologist; however; diagnosis of (CH) and (AH) made by other pathologists were not concordant with morphometric data.

Conclusion

It may be necessary to make some revisions in WHO classification for endometrial hyperplasia and precancerous lesions.     

Computerized interactive morphometry as an aid in the diagnosis of pleural effusions

A morphometric study of cytologic preparations from patients with benign and malignant (mesothelioma and carcinoma) pleural effusions is reported. The routine cytologic smears from these specimens were studied with a new system of video-based computerized interactive morphometry (CIM) that allows the measurements of real-time images of cell profiles by the simple procedure of touching the two extreme points of a diameter of interest on a touch-sensitive screen. For each cell, the nuclear profile diameter (NPD) and the cytoplasmic profile diameter (CPD) are measured and categorized into classes with 2-microns intervals; the NPD/CPD ratio is also calculated. The mean NPD is calculated for the specimen after measurement of 100 cells. The data were interpreted by two independent methods: a statistical method of discriminant analysis that classifies the lesions as benign, carcinoma or mesothelioma and provides a probability statement of membership in a particular diagnostic class and an ad-hoc algorithm that categorizes the effusions as benign or malignant based on hierarchic analysis. A data base derived from study of the first 24 cases was constructed and utilized for the test classification of the second 24 cases, which were treated as specimens of unknown diagnosis. The discriminant analysis correctly classified 21 of the 24 test cases into their proper diagnostic groups. The algorithm for a computer-generated pathologic diagnosis correctly identified 47 of the 48 cases as benign or malignant. The technical advantages of video-based CIM over the existing morphometric methods are discussed.     

The crowding effect and morphometric variability in Echinostoma caproni (Digenea: Echinostomatidae) from ICR mice

Population density, or crowding, was examined to determine its effect on the morphometric variability of Echinostoma caproni (Digenea) in ICR mice. Six mice were infected with 25 and 100 metacercariae, and a single mouse was infected with 300 metacercariae. All mice were infected at necropsy 22 days postinfection with recoveries of 77%, 69%, and 7.3%, respectively. Whole mounts were prepared, and 31 characters were evaluated (25 direct measurements and 6 ratios). Univariate and multivariate statistical analysis revealed significant differences between adult worms from all 3 groups. Twenty-seven of 31 characters showed significant within-group differences, with the primary differences between worms from 25/100 versus 300 metacercariae infections. Discriminant function analysis yielded a 100% correct classification based on infection size, which is consistent with studies on distinct species of Echinostoma. The low recovery from the mouse infected with 300 metacercariae suggests inflammatory expulsion of juvenile worms and the possibility of immunity as a factor in the crowding effect. These results suggest that external factors may affect morphometric variability of digenetic trematodes to a larger degree than previously recognized.     

Identification of components of fibroadenoma in cytology preparations using texture analysis: a morphometric study

S. Singh and R. Gupta Identification of components of fibroadenoma in cytology preparations using texture analysis: a morphometric study Objectives: To evaluate the utility of image analysis using textural parameters obtained from a co‐occurrence matrix in differentiating the three components of fibroadenoma of the breast, in fine needle aspirate smears. Methods: Sixty cases of histologically proven fibroadenoma were included in this study. Of these, 40 cases were used as a training set and 20 cases were taken as a test set for the discriminant analysis. Digital images were acquired from cytological preparations of all the cases and three components of fibroadenoma (namely, monolayered cell clusters, stromal fragments and background with bare nuclei) were selected for image analysis. A co‐occurrence matrix was generated and a texture parameter vector (sum mean, energy, entropy, contrast, cluster tendency and homogeneity) was calculated for each pixel. The percentage of pixels correctly classified to a component of fibroadenoma on discriminant analysis was noted. Results: The textural parameters, when considered in isolation, showed considerable overlap in their values of the three cytological components of fibroadenoma. However, the stepwise discriminant analysis revealed that all six textural parameters contributed significantly to the discriminant functions. Discriminant analysis using all the six parameters showed that the numbers of pixels correctly classified in training and tests sets were 96.7% and 93.0%, respectively. Conclusion: Textural analysis using a co‐occurrence matrix appears to be useful in differentiating the three cytological components of fibroadenoma. These results could further be utilized in developing algorithms for image segmentation and automated diagnosis, but need to be confirmed in further studies.     

Environmental factors influencing zooplankton species composition of lakes in north-central Ontario,Canada

To assess the relative importance of lake chemistry, morphometry and zoogeography on limnetic zooplankton, we collected zooplankton, water, and morphometric data from 132 headwater Canadian Shield lakes in 6 regions across north-central Ontario. A subset of these lakes (n = 52) were fished with gill nets. We clustered lakes based on their zooplankton species composition (presence/absence). Discriminant analysis was employed to determine how well lake characteristics could predict zooplankton community types. Correct classification of zooplankton communities for three models ranged from 72 to 91%. Lake size, lake location, and buffering capacity were ranked as the most important factors separating lake groups. Fish abundance (CPUE) was not significant in distinguishing between zooplankton communities. Though the range of lake sizes was limited (1–110 ha), larger lakes tended to support more species. Lake location (zoogeography) also influenced species composition patterns. Although Algoma lakes tended to be larger (\-x = 18.0 ha, other lakes \-x = 2.5 ha), they supported relatively depauperate zooplankton communities. Buffering capacity was ranked third in the discriminant analysis models, but pH and alkalinity were not significantly different between lake groups.     

Taxonomic re-evaluation of the jaguar

The purpose of this report is to re-examine jaguar (Panthera onca) subspecies using morphometric skull characters and multivariate statistical methods to analyze differences. Eleven skull characters used historically to define jaguar subspecies were measured on 170 specimens in U.S. natural history museums. Relationships among the eight recognized subspecies of jaguar were examined. using Multigroup Discriminant Analysis (MDA) and univarite statistics. Discriminant analyses of these skull characteristics does not indicate distinct groups, supporting the null hypothesis. This analysis was conducted to add to systematic studies such as DNA analysis to assist the Felid Taxon Advisory Group (Felid TAG) and the American Zoo and Aquarium Association (AZA) establish valid taxonomic differences in the jaguar. Zoo Biol 16:107–120, 1997. © 1997 Wiley-Liss, Inc.     

Separation of murine neutrophils and macrophages by thermoresponsive magnetic nanoparticles

Magnetic particles have been used widely in both biotechnological and medical fields, including for immunoassay, enzyme immobilization, drug transport, and immunological diagnosis. Especially particles with bioactive molecules such as antibodies and streptavidin are very useful tools for cell separation. Here we report affinity selection of neutrophils and macrophages from peritoneal inflammatory cells performed by thermoresponsive magnetic nanoparticles conjugated with macrophage-specific anti-F4/80 antibody. The magnetic nanoparticles, which are capped with thermoresponsive polymers, are aggregated by heating the particles over 30 degrees C and show their intrinsic magnetism. The neutrophils are concentrated approximately 90% by these magnetic nanoparticles without any activation, indicating that this novel cell separation method could fulfill a wide range of applications in analysis of the isolation of fragile cells such as neutrophils.     

Morphometric analysis of Alaskan members of the genus Potentilla sect. Niveae (Rosaceae)

A morphometric study of Potentilla nivea, P unijlora, and P hookeriana, as well as the close relative of the latter, P furcata, has been carried out, and the quantitative data subjected to Canonical Discriminant Analysis. The four taxa belong to the arctic-alpine section Niveae of Potentilla, and material for the analysis was collected in Alaska, U. S. A. The a priori defined groups are based on petiole hair type, the qualitative, and only, character traditionally used to distinguish taxa within Potentilla sect. Niveae. The hair types recognized previously by taxonomists have been vaguely defined, and the intraspecific variation of other morphological characters has never been discussed. Ordination by canonical discriminant analysis was performed to characterize mean differences among species, to obtain insight into group differences, and to estimate character weights from correlations between canonical variates and original variables. The four taxa differ significantly in the canonical analysis of six quantitative characters. Leaflet length, incision depth (length of leaflet teeth), and ovule number are shown to be the most important discriminators. A key to the four taxa, taking into account the intra- vesus interspecific variation, as well as character weights, is provided.