[Unilateral peromelia of the left pelvic limb in a Brown Swiss calf]. / Unilaterale Peromelie der linken Beckengliedmasse bei einem Brown Swiss Kalb.

INTRODUCTION: Congenital deformities of the limbs occur sporadically in various species, but the cause is often unclear. The clinically healthy female Brown Swiss calf presented here showed a congenital peromelia of the left hind limb. The affected limb is twisted, disproportional and the bones distally of the metatarsus are missing. Karyotyping and genome sequencing did not indicate on a genetic cause of the anomaly. An infection with the Schmallenberg virus could not be ruled out. Furthermore, there was no evidence of further adverse environmental effects during pregnancy.

INTRODUCTION: Des malformations congénitales des membres, dont la cause est souvent peu claire, surviennent sporadiquement chez diverses espèces. Le veau Brown Swiss femelle présenté ici, tout en étant cliniquement sain, présentait une péromélie congénitale du postérieur gauche. Le membre concerné été en rotation interne, disproportionné et les os distalement au métatarse étaient absents. La détermination du caryotype et le séquençage de l'ensemble du génome n'ont apporté aucun élément parlant pour une cause génétique de l'anomalie. Il n'a pas été possible d'exclure une infection par le virus de ­Schmallenberg. D'autre part il n'y avait aucun élément évoquant d'autres influences environnementales néfastes durant la gestation.

Automated detection of cancer cells in effusion specimens by DNA karyometry.

BACKGROUND: The average sensitivity of conventional cytology for the identification of cancer cells in effusion specimens is only approximately 58%. DNA image cytometry (DNA-ICM), which exploits the DNA content of morphologically suspicious nuclei measured on digital images, has a sensitivity of up to 91% for the detection of cancer cells. However, when performed manually, to our knowledge to date, an expert needs approximately 60 minutes for the analysis of a single slide. METHODS: In the current study, the authors present a novel method of supervised machine learning for the automated identification of morphologically suspicious mesothelial and epithelial nuclei in Feulgen-stained effusion specimens. The authors compared this with manual DNA-ICM and a gold standard cytological diagnosis for 121 cases. Furthermore, the authors retrospectively analyzed whether the amount of morphometrically abnormal mesothelial or epithelial nuclei detected by the digital classifier could be used as an additional diagnostic marker. RESULTS: The presented semiautomated DNA karyometric solution identified more diagnostically relevant abnormal nuclei compared with manual DNA-ICM, which led to a higher sensitivity (76.4% vs 68.5%) at a specificity of 100%. The ratio between digitally abnormal and all mesothelial nuclei was found to identify cancer cell-positive slides at 100% sensitivity and 70% specificity. The time effort for an expert therefore is reduced to the verification of a few nuclei with exceeding DNA content, which to our knowledge can be accomplished within 5 minutes. CONCLUSIONS: The authors have created and validated a computer-assisted bimodal karyometric approach for which both nuclear morphology and DNA are quantified from a Feulgen-stained slide. DNA karyometry thus increases the diagnostic accuracy and reduces the workload of an expert when compared with manual DNA-ICM.

Interfertility and genetic variability among European and North American isolates of the basidiomycete fungus Chondrostereum purpureum.

The conspecificity of Finnish and western Canadian isolates of the decay fungus Chondrostereum purpureum was investigated by several approaches, including the assessment of genetic variability, mating and progeny analysis, and the analysis of selected phenotypic traits. Eight second-generation single spore strains per fungal isolate pairing were investigated with specific genetic markers developed for both Finnish and Canadian parental isolates. Tests of linkage disequilibrium were used to analyze whether these markers assorted independently among single spore strains. This procedure was similarly applied to the third-generation spore progeny. Finally, global non-metric multidimensional scaling was used to analyze independent random amplified microsatellite marker data to assess the genetic variability of the parental Finnish and Canadian isolates, and their second- and third-generation progeny. Our results revealed that the parental isolates from Finland and western Canada were genetically divergent, but no interfertility barriers were identified between these geographically distant fungi. Furthermore, parental genetic markers used in mating studies demonstrated that second- and third-generation spore progenies underwent normal meiosis and genetic recombination without linkage disequilibrium. Based on this work, the studied C. purpureum isolates from Finland and Canada can be considered as belonging to a single biological species, although genetic and limited phenotypic differentiation was observed.

[INFLUENCE OF AGGRESSION AND RELATED PSYCHOLOGICAL CHARACTERISTICS OF ATHLETES IN THE FREQUENCY OF NUCLEAR VIOLATIONS IN EXFOLIATED CELLS OF THE ORAL CAVITY].

The frequency of cells with abnormal nuclear morphology (micronuclei, perinuclear vacuo-les, notches, protrusions, such as «tongue¼ and «broken egg¼) in the buccal epithelium of wrestlers on different days of the competition period was identified. The largest number of violations observed on the 3rd day after the competition. It was conducted psychological testing of athletes and it was determined 16 psychological characteristics associated with the aggressiveness of athletes. It was revealed the effect of psychoemotional state on the cytogenetic apparatus athletes. It was found associations of the dynamics of reactive anxiety athletes and frequency of nuclear aber- 327 rations. The hypothetical scheme of influence human aggression and related psychological characteristics of its genetic apparatus by neurohumoral system was constructed.

[Karyometry of BBN-induced precancerosis of the urothelium : An experimental analysis]. / Karyometrie BBN-induzierter Präkanzerosen des Urothels : Eine experimentelle Analyse.

BACKGROUND: The morphology of experimental precancerous lesions of the urinary bladder has been interpreted quite differently by various authors. OBJECTIVES: The aim of this investigation was to quantify these lesions by karyometry and, thus, to gain a more reliable understanding of the process. MATERIALS AND METHODS: A total of 60 Wistar rats were fed with N­butyl-N-(4-hydroxybutyl)nitrosamine (BBN) at a concentration of 0.05 % in their drinking water to induce preneoplastic changes of the urothelium. After the second week of BBN exposition, 6 animals were killed every 2 weeks up to week 20. Smears of the scraped off urothelium of 3 urinary bladders of each group were analyzed cytologically and karyometrically. RESULTS: BBN exposition led to statistically significant changes of the karyometric values using the χ2 test to differentiate the control animals from the ones that had ingested BBN and the 2­week groups from each other. These changes consisted mainly in significant deviations of the size of the nuclear area within the different groups. CONCLUSION: Morphological and karyometrical analysis showed that biologically relevant stages in the development of chemically induced urothelial precancerous lesions could be realized much earlier than had been assumed in recent publications. Karyometric analysis offered a valid basis to describe the early morphologic alterations of carcinogenesis.

Severe dysplasia can be distinguished from moderate and mild dysplasia of bronchial mucosa by changes in Ki-67 index.

Preneoplastic lesions on small bronchial biopsy specimens may cause a diagnostic dilemma. The aim of this study was to estimate karyometric variables and the Ki-67 index of preneoplastic bronchial lesions and squamous cell carcinoma of the lung. The study was performed on endoscopic samples of squamous cell carcinoma (n = 22), normal appearing mucosa surrounding carcinoma (n = 10), bronchial dysplasia of mild (n = 7), moderate (n = 6), and severe grade (n = 6), carcinoma in situ (n = 17), and normal mucosa from patients with chronic bronchitis (n = 26). Karyometric analysis was done using the image analyzer ImageJ 1.47q. Ki-67 activity was also quantified by ImageJ 1.47q with the plugin Cell Counter. The highest values of nuclear area were found in squamous cell carcinoma, and differences were statistically significant compared to normal mucosa, all grades of dysplasia and normal appearing mucosa surrounding carcinoma (p < 0.01). The Ki-67 index was significantly higher in squamous cell lung carcinoma compared to normal mucosa, mild and moderate dysplasia and normal appearing mucosa surrounding carcinoma (p < 0.01). The Ki-67 index was significantly higher in severe dysplasia than in mild and moderate dysplasia (p < 0.01). In conclusion, the Ki-67 index is a useful parameter for more objective grading and can be of prognostic value to determine the biological potential of preneoplastic bronchial lesions.

High Proportion of Nuclear Phenotype Identifies Aggressive Cutaneous Squamous Cell Carcinoma.

OBJECTIVE: To develop a quantitative histopathology algorithm to predict which patients with cutaneous squamous cell carcinoma (cSCC) were likely to experience recurrence or metastases. STUDY DESIGN: This retrospective study of cSCC lesions compared patients with aggressive disease (n = 40) and those with nonaggressive disease (n = 35). Based on a previous study using nuclear karyometry, we determined that aggressive lesions had a high proportion of a specific nuclear phenotype. The proportion of those nuclei was used to derive an aggressiveness score for each lesion. The mean age of patients was similar in both groups, as were the locations of index lesions. RESULTS: The mean aggressiveness scorefor cases with aggressive lesions was 0.60 ± 0.21 and was 0.28 ± 0.35 for those with nonaggressive lesions. The overall accuracy in properly characterizing lesions was 72%. The area under the receiver operating characteristic curve was 0.80 ± 0.05. In general, the aggressive nuclear phenotype is represented by elevated levels of chromatin clumps and short linear segments of dark chromatin/intense pixels. CONCLUSION: These data suggest that discriminant functions may be utilized to distinguish between aggressive and nonaggressive lesions at the time of diagnosis.

Classification in karyometry: performance testing and prediction error.

Classification plays a central role in quantitative histopathology. Success is expressed in terms of the accuracy of prediction for the classification of future data points and an estimate of the prediction error. The prediction error is affected by the chosen procedure, e.g., the use of a training set of data points, a validation set, an independent test set, the sample size and the learning curve of the classification algorithm. For small samples procedures such as the "jackknife," the "leave one out" and the "bootstrap" are recommended in order to arrive at an unbiased estimate of the true prediction error. All of the procedures rest on the assumption that the data set used to derive a classification rule is representative for the diagnostic categories involved. It is this assumption that in quantitative histopathology has to be carefully verified before a clinically generally valid classification procedure can be claimed.

Effect of statins and aerobic physical exercise on liver function in dyslipidemic rats: morphometric study / Efecto de las estatinas y ejercicio físico aeróbico sobre la función hepática en ratas dislipémicas: estudio morfométrico

The present study was carried out to evaluate the effect of statins associated with physical exercise (PE) in liver cells in dyslipidemic rats through cariometry. The animals were divided into six groups: animals subjected to a hypercholesterolemic diet (HD), simvastatin, with (G1) and without (G2) physical exercise (PE); HD submitted (G3) or not (G4) to PE, and commercial food diet (F) with (G5) and without (G6) PE. Histological analysis of the liver was performed by staining the slides with hematoxylin and eosin. The cariometric study included measuring the major and minor diameters of the hepatocytes nuclei. The Shapiro-Wilk test was also performed. To determine the differences among the groups, the Kruskal-Wallis Test with Dunn's post-test were conducted. The significance level was set at 5 percent. No difference was found in the hepatocytes nuclei between G5 and G6. When these groups were related with G3 and G4, reduced nuclei were observed. There was no difference between G1 and G6. The comparison between G6 and G2 showed that the nuclei in G2 were smaller. No difference was detected between G5 and G1. Changes were observed in the nuclei shape in G2 in comparison to G1. Considering G2 and G3, a decrease in the size of nuclei was observed in G3. On the other hand, G2 showed changes in shape in the comparative analysis with G4. The size and shape of G1 nuclei were larger than G3 as well as changes in shape were observed when compared to G4. G4 showed smaller nuclei than G3. Therefore, F, associated or not with the practice of PE, does not alter the size and shape of the hepatocytes nuclei; HD combined with sedentarism influences changes in the morphometric parameters of hepatocytes; and the association of simvastatin and PE seems to protect the hepatocytes nuclei with regard to HD.

El presente estudio se realizó con la finalidad de evaluar el efecto de las estatinas asociadas con el ejercicio físico (PE) en las células del hígado, en ratas con dislipidemia a través de cariometría. Los animales fueron divididos en seis grupos: animales sometidos a una dieta hipocolesterolemiante (HD), simvastatina, con (G1) y sin (G2) ejercicio físico (PE); HD enviado (G3) o no (G4) para educación física y dieta comercial (F) con (G5) y sin (G6) PE. El análisis histológico del hígado se realizó por tinción de los portaobjetos con hematoxilina y eosina. El estudio cariométrico incluyó la medición de los diámetros mayor y menor de los núcleos de hepatocitos. Se realizó la prueba de Shapiro-Wilk. Para determinar las diferencias entre los grupos, se realizó la prueba de Kruskal-Wallis con Dunn. El nivel de significación se fijó en 5 por ciento. No se encontraron diferencias en los núcleos de hepatocitos entre G5 y G6. Los núcleos fueron observados cuando estos grupos estaban relacionados con G3 y G4. No hubo diferencia entre G1 y G6. La comparación entre G6 y G2 mostró que los núcleos en G2 eran más pequeñas. No se detectaron diferencias entre el G5 y G1. Se observaron cambios en la forma núcleos en G2 en comparación con G1. Considerando G2 y G3, se observó en G3 una disminución en el tamaño de los núcleos. En el análisis comparativo con G4, G2 mostró cambios en la forma . El tamaño y forma de los núcleos G1 eran más grandes que G3, así como cambios en la forma se observaron cuando se compararó con G4. G4 mostraron núcleos menores que G3. Por tanto, F, asociados o no a la práctica de PE, no altera el tamaño y la forma de los núcleos de hepatocitos; HD combinada con influencias sedentarismo cambios en los parámetros morfométricos de los hepatocitos, y la asociación de simvastatina y PE parece proteger a los hepatocitos con respecto a la HD.

[Basic aspects of medical genetics]. / Grundlagen der medizinischen Genetik.

The human genome consists of 23 pairs of chromosomes that contain 20 000-25 000 genes. Genetic disorders can be caused by different mechanisms, and therefore the confirmation of a suspected diagnosis requires knowledge of the underlying defect, so that the correct test can be applied. Monogenic diseases are caused by disturbances in a single gene, and currently only targeted diagnostic testing is available following a specific clinical suspicion. Chromosomal disorders usually involve multiple genes, so that the symptoms are often less specific. Specialists in Medical Genetics FMH are trained in creating a clinical genetic differential diagnosis, requesting the according laboratory test, interpretating the results and providing expert genetic counseling in presymptomatic and prenatal diagnosis. In Switzerland, specific legal principles and ethical guidelines must be taken into account.

Le génome humain est constitué de 23 paires de chromosomes et contient 20 000 à 25 000 gènes. Les maladies génétiques peuvent être causées par différents mécanismes, or pour confirmer un diagnostic présumé et utiliser le test adéquat, il est nécessaire de connaître le défaut génétique sous-jacent. Les maladies monogéniques sont causées par des perturbations dans un seul gène, et actuellement seul un test diagnostique ciblé peut être réalisé suite à une suspicion clinique spécifique. Les anomalies chromosomiques impliquent généralement plusieurs gènes, donc les symptômes sont souvent moins spécifiques. Les spécialistes en génétique médicale FMH sont formés à reconnaître cliniquement les diagnostics génétiques, à demander les analyses correspondantes, à en interpréter les résultats et à donner un conseil génétique adapté en diagnostic présymptomatique et prénatal. En Suisse, des principes juridiques et éthiques spécifiques doivent également être pris en compte.

Karyometry and quantitative immunohistochemical analysis of the urothelium in tissue sections: a feasibility study based on chromatin remodeler DAXX immunostaining.

The aim of this study was to investigate the feasibility of applying a software traditionally used in the field of engineering to pathology, in particular to tissue sections from normal urothelium (NU) immuno-stained for the chromatin remodeler DAXX (death domain-associated protein). The study included 5 cases of NU. Images were recorded with a Nikon digital camera. The nuclear area and the intensity of nuclear staining were analyzed with a software package developed in LabVIEW environment. The nuclear size is 14.8 plus or minus 6.5 square microns. The nuclei in the cells adjacent to the stroma are slightly smaller than in the intermediate cells by a factor of 0.86. The mean nuclear area of the nuclei in the superficial cell layer in NU is identical to the nuclei in the intermediate cell layers. For each nucleus intensity of nuclear staining is calculated based on the gray value of the individual picture elements in the green color plane. The mean and standard deviation of nuclear gray value are 106 plus or minus 15. The mean value in the nuclei adjacent to the stroma is slightly greater by a factor 1.02 and 1.04 compared to the intermediate and superficial cell layers. In conclusion, this exploratory study shows that karyometry and quantitative immunohistochemical analysis can be done accurately by using a digital camera commonly available to pathologists and an image analysis software routinely used in the field of engineering.

Estresse pós-privação de sono: análises morfométricas do fígado / Stress caused by sleep deprival: liver morphometric analysis

OBJETIVO: Avaliar o efeito da exposição ao estresse pós-privação de sono em características celulares e nucleares do fígado de ratos. MÉTODOS: 16 ratos (Wistar) machos adultos (200-260 g) foram mantidos em ciclo de luz controlado recebendo dieta com quantidades usuais de sal e livre acesso à água e alimento. Os animais foram divididos em dois grupos de oito animais (grupo experimental e grupo controle). Os animais permaneceram na mesma gaiola (dois de cada vez) durante sete dias e, após esse período, foram pesados e separados. O animal do grupo controle continuou na mesma gaiola e o animal do grupo experimental foi colocado em aparato de privação de sono. Após as 96 horas, os dois ratos foram pesados,anestesiados, sacrificados com dose excessiva de anestésico e os fígados foram retirados. RESULTADOS: Não há diferença significante entre o grupo experimental e grupo controle em relação ao peso, embora haja diminuição de peso no grupo experimental. Nas análises cariométricas, houve diferença significante em relação ao diâmetro menor (p=0,03) e volume (p=0,04) do núcleo dos hepatócitos do grupo experimental. Nas análises esteriológicas, houve diferença significante do grupo experimental no núcleo (maior, p=0,036), do citoplasma (menor p=0,009) e outras estruturas (maior p=0,008). CONCLUSÃO: O estresse parece contribuir para alteração na estrutura celular hepática.

OBJECTIVE: To evaluate the effect of stress induced by sleep deprivation on nuclear and cellular features of rat liver. METHODS: 16 adult male Wistar rats (200-260 g) in controlled light cycle received diet with customary salt quantity and free access to water and food. The animals were divided into two groups with 8 animals each (experimental group and control group). Animals stayed in the same box (two at a time) during seven days andafter that period they were weighted e separated. The control group animal continued in the same box and the experimental group animal were transferred to a sleep deprivation apparatus. After the 96 hours the animals were sacrificed by an excessive anesthetic dose; animals were weighted and their liver extracted. RESULTS: There was no significant difference between experimental and control group regarding weight, although therewas a decrease of weight on the experiment group. Karyotipical analysis showed significant smaller diameter (p=0.03) and volume (p=0.044) of hepatocyte nuclei in experimental group. Stereological analyses showed significant differences in experimental group for nucleus (larger, p=0.036), cytoplasm (smaller, p=0.009) and other structures (larger, p=0.008). CONCLUSION: Stress seems to contribute to alteration of hepatic cell structure.

A randomized phase IIb presurgical study of finasteride vs. low-dose flutamide vs. placebo in men with prostate cancer. Efficacy monitored by karyometry.

OBJECTIVE: Presurgical, window of opportunity trials have been proposed as a model to assess the activity of preventive and therapeutic interventions in a cost-effective manner in prostate cancer (CaP). The aim of the study was to explore karyometry as a method for monitoring the efficacy of intervention with preventive agents in patients with CaP. MATERIALS AND METHODS: The material used in this investigation was from the 2F study, i.e., an Italian prospective randomized phase IIb presurgical study of finasteride vs. low-dose flutamide vs. placebo in men with CaP. Image analysis was performed in 16 cases treated with finasteride, 24 with flutamide, and 20 with placebo. For all these cases, CaP and normal looking secretory epithelium were present in the pretreatment biopsies as well as the post-treatment ex-vivo biopsies obtained from the radical prostatectomy specimens. RESULTS: To establish a direction of nuclear change from normal to malignancy, i.e., the so-called line of progression, a discriminant function was derived with the normal looking epithelium in the pretreatment biopsies as one endpoint, and the CaP in the pretreatment biopsies as the other. The discriminant function was then applied to the post-treatment groups. The increase in relative nuclear area was the dominant feature. In the placebo group, 15 out of 20 CaP (75%) cases had a higher discriminant function score at the end of study, with a significant increase of the mean score by 90%. The flutamide treated CaP cases had increased discriminant function scores in 19 out of 24 cases (79%) and an increase of the mean score by 43%; the 5 cases with lower scores involved only minor reductions. In contrast, the finasteride treated CaP cases had increased discriminant function scores for 8 out of 16 cases (50%), but the increase in the mean score was by only 8%. CONCLUSION: This exploratory study establishes that karyometric monitoring can track the results of subtle nuclear changes induced by preventive interventions in men with CaP, thus allowing assessment of agent activity in a cost-effective manner.

Differential diagnosis of follicular tumor by expert systems based on a set of quantitative features of thyrocyte nuclei and aggregates.

OBJECTIVE: To develop expert systems for classification of follicular thyroid tumor at a preoperative stage. STUDY DESIGN: Fine needle aspiration biopsy of the thyroid gland with a histologic conclusion of follicular cancer and follicular adenoma were the object of the morphometric study. General sample size was 4500 nuclei and 3000 aggregates. RESULTS: Quantitative regularities of pathologic changes in thyrocyte nuclei and aggregates in follicular cancer and follicular adenoma were revealed. Threshold values and weighting coefficients of quantitative features of thyrocyte nuclei and aggregates characterizing cancer made the basis of the two expert systems. Expert systems included standard 2-D S-matrix containing threshold values of nuclei and aggregates in cancer and their weighting coefficients as well as 1-D scientific X-matrix designed for filling with quantitative features of the studied object. The diagnosis was verified by the value of a diagnostic index by means of comparing feature values in the corresponding elements of S- and X-matrices. After that, a diagnostic index was calculated taking into account the features' weighting coefficient. CONCLUSION: The developed expert systems based on a set of quantitative features of thyrocyte nuclei and aggregates will allow assessing the malignant potential of a follicular thyroid tumor at a preoperative stage.

Karyometric comparison of splenic and gastric marginal zone lymphomas.

BACKGROUND: Marginal zone lymphomas are indolent B-cell lymphomas associated with autoimmunity and chronic inflammation. The two most frequent variants are mucosa associated lymphoid tissues marginal zone lymphomas and splenic marginal zone lymphomas. The aim of the study was to determine if it is possible to classify splenic and gastric lymphomas according to karyometric features. METHODS: The material consisted of 16 splenic and 14 gastric lymphomas. The measurements were done with the AnalySIS image analysis system. In each case at least 100 nuclei were selected, and 19 different geometric parameters were measured. RESULTS: On statistical analysis, the nuclei of splenic and gastric lymphomas showed differences in most parameters, but significant overlap of the values was present. Neural networks were trained and used for classification of the data. By this method, the nuclei were properly classified with a sensitivity of 0.75 and specificity of 0.71. In addition, in all the cases the majority of the nuclei were properly classified, thus allowing correct classification of all the cases into "splenic" or "gastric". CONCLUSION: These results support the view that mucosa-associated lymphoid tissue lymphomas and splenic marginal-zone lymphomas are separate entities.

Karyometry of nuclear phenotypes in cutaneous squamous cell cancer.

OBJECTIVE: To establish the karyometric characteristics of the two main nuclear phenotypes in cutaneous squamous cell cancer (cSCC) lesions. STUDY DESIGN: The clinical materials comprised 75 cases of cSCC, 38 with aggressive lesions and 37 with nonaggressive lesions. High-resolution images of 100 nuclei per case were recorded. Data were partitioned into four subgroups covering the range of lesion progression. Four discriminant functions were derived to distinguish aggressive from nonaggressive lesions. The most typical nuclei from the phenotype predominant in aggressive lesions and nonaggressive lesions were separated out by thresholding on the discriminant function score axes. For these homogeneous sets of nuclei the karyometric features were computed. RESULTS: The nuclear populations in cSCC lesions are a very heterogeneous set. There are two axes of dispersion, along the line of lesion progression and between aggressive and nonaggressive lesions. The analysis faces the difficulty that lesions from both diagnostic categories contain nuclei of the same two phenotypes with the difference between categories consisting only of differences in proportion of the two phenotypes. CONCLUSION: The nuclei of the aggressive phenotype I and nonaggressive phenotype II have substantially different chromatin patterns and can be distinguished with > 90% correct recognition rate.

Karyometry in atypical endometrial hyperplasia: a Gynecologic Oncology Group study.

OBJECTIVES: Treatment for atypical endometrial hyperplasia (AEH) is based on pathologic diagnosis. About 40% of AEH is found to be carcinoma at surgery. This study's objective is to derive an objective characterization of nuclei from cases diagnosed as AEH or superficially invasive endometrial cancer (SIEC). METHODS: Cases from GOG study 167A were classified by a central pathology committee as AEH (n=39) or SIEC (n=39). High resolution digitized images of cell nuclei were recorded. Features of the nuclear chromatin pattern were computed. Classification rules were derived by discriminant analysis. RESULTS: Nuclei from cases of AEH and SIEC occupy the same range on a progression curve for endometrial lesions. Cases of AEH and SIEC both comprise nuclei of two phenotypes: hyperplastic characteristics and premalignant/neoplastic characteristics. The principal difference between AEH and SIEC is the percentage of premalignant/neoplastic nuclei. When this percentage approaches 50-60% superficial invasion is likely. SIEC may develop already from lesions at the low end of the progression curve. CONCLUSIONS: AEH comprises cases which may constitute a low risk group involving <40% of AEH cases. These cases hold a percentage of <20% of nuclei of a preneoplastic phenotype. AEH cases from the central and high end of progression have >40% of nuclei of preneoplastic phenotype. Nuclei of the preneoplastic phenotype in AEH lesions are almost indistinguishable from nuclei in SIEC, where this percentage exceeds 60%. The percentage of nuclei of the preneoplastic phenotype in AEH esions might serve as criterion for assessment of risk for the development of invasive disease.

[Results of karyometry of acinar adenocarcinoma of the prostate].

The paper presents the results of karyometry of acinar adenocarcinoma of the prostate in the biopsy specimens with total Gleason scores of 6, 7, and 8-10. There are statistically significant differences in the morphometric characteristics of tumor cell nuclei (the long and short diameters of nuclei and nucleoli, their area and perimeter) in the biopsy specimens as compared with those with total Gleason scores of 6 and 7, as well as in those of the nucleoli as compared to those with total Gleason scores of 6 and 7 and 7 and 8-10. The findings suggest that there are karyometric differences in the cells of prostatic acinar adenocarcinoma in the groups with total Gleason scores of 6, 7, and 8-10.

Automation of the buccal micronucleus cytome assay using laser scanning cytometry.

Laser scanning cytometry (LSC) can be used to quantify the fluorescence intensity or laser light loss (absorbance) of localized molecular targets within nuclear and cytoplasmic structures of cells while maintaining the morphological features of the examined tissue. It was aimed to develop an automated LSC protocol to study cellular and nuclear anomalies and DNA damage events in human buccal mucosal cells. Since the buccal micronucleus cytome assay has been used to measure biomarkers of DNA damage (micronuclei and/or nuclear buds), cytokinesis defects (binucleated cells), proliferative potential (basal cell frequency), and/or cell death (condensed chromatin, karyorrhexis, and pyknotic and karyolytic cells), the following automated LSC protocol describes scoring criteria for these same parameters using an automated imaging LSC. In this automated LSC assay, cells derived from the buccal mucosa were harvested from the inside of patient's mouths using a small-headed toothbrush. The cells were washed to remove any debris and/or bacteria, and a single-cell suspension prepared and applied to a microscope slide using a cytocentrifuge. Cells were fixed and stained with Feulgen and Light Green stain allowing both chromatic and fluorescent analysis to be undertaken simultaneously with the use of an LSC.