Functional role of ALK-related signal cascades on modulation of epithelial-mesenchymal transition and apoptosis in uterine carcinosarcoma.

BACKGROUND: Anaplastic lymphoma kinase (ALK), which is a receptor tyrosine kinase, is essentially and transiently expressed in the developing nervous system. Recently, the deregulated expression of full-length ALK has been observed in some primary solid tumors, but little is known about its involvement in the tumorigenesis of uterine carcinosarcomas (UCSs). Here we examined the functional role of the ALK gene in UCSs. METHODS: Regulation and function of the ALK gene were assessed using two endometrial carcinoma cell lines. Expression of ALK and its related molecules were also investigated using clinical samples of UCSs. RESULTS: In cell lines, ALK promoter activity was significantly increased by transfection of Sox11 and N-myc, which are known to contribute to neuronal properties. Cells stably overexpressing full-length ALK showed an enhancement of EMT properties mediated by TGF-ß1 and HGF, along with an increase in phosphorylated (p) Akt and nuclear p65. Overexpression of p65 also led to transactivation of Twist1 gene, known as an EMT inducer. Finally, treatment of the stable ALK-overexpressing cells with doxorubicin resulted in inhibition of apoptosis with progressive increase in the expression ratio of both pAkt and bcl2 relative to total Akt and bax, respectively. In clinical samples, strong cytoplasmic ALK immunoreactivity and mRNA signals without rearrangement or amplification of the ALK locus were frequently observed in UCSs, particularly in the sarcomatous components. Further, ALK IHC score was found to be positively correlated with Sox11, N-myc, Twist1, and bcl2 scores. CONCLUSION: ALK-related signal cascades containing Akt, NF-κB, Twist1, and bcl2 may participate in initial signaling for divergent sarcomatous differentiation driven from carcinomatous components in UCSs through induction of the EMT process and inhibition of apoptotic features.

Action of physiologically active materials in human semen during aging.

We examined the activities of some physiologically active substances (enzymes) in human seminal plasma from subjects in their 20s, 30s, and 40s. The average volume of semen per ejaculation in this investigation did not differ significantly depending on the age of the subject. Two age dependent patterns of decrease of substances in semen were observed, and the substances including tissue kallikrein and prostate specific antigen (PSA) basic arginine amidase in human seminal plasma showing the first pattern (a significant decrease in the 40s as compared to the 30s) might be initially secreted from the prostate gland, and whereas the glands secreting the other group of substances including active form coagulation factor X (FXa) and plasminogen are not now known. The levels of these substances in semen decrease in the subjects in their 30s. The coagulation and liquefaction times of human semen from older subjects were both prolonged with those of semen from younger subjects, and that such alteractions ultimately cause the age dependent declines of the motility of sperm and the ability of fertility.

[A case of systemic lupus erythematosus with hemophagocytic syndrome and cytophagic histiocytic panniculitis].

A 23-year-old man, admitted because of high fever, polyarthralgia, butterfly rash and chest pain, was diagnosed as systemic lupus erythematosus (SLE) from the findings of positive antinuclear antibody and anti-DNA antibody. He was treated with 60 mg prednisolone daily, but as reducing the dose, white blood cell counts and platelet counts were decreased and fever, polyarthralgia, decrease of complements, increase of ferritin, hepato-splenomegaly and liver dysfunction were observed. Bone marrow specimen revealed phagocytosis of blood cells by histiocytes and he was diagnosed as hemophagocytic syndrome(HPS) due to active SLE. Methylprednisolone pulse therapy was effective temporarily, HPS recurred while reducing steroid, and cyclosporin was added. After a temporary remission, marked extensive swelling in the face appeared suddenly. Facial skin biopsy showed necrosis of fat cells and hemophagocytosis by histiocytes. Accordingly, he was diagnosed as panniculitis due to HPS and was treated successfully with intravenous cyclophosphamide pulse therapy and high dose of gammaglobulin. Several cases of HPS due to SLE have been reported recently, but this is a rare case of cytophagic histiocytic panniculitis (CHP) due to SLE.

Genes encoding nitrilase-like proteins from tobacco.

Nitrilase (nitrile aminohydrolase, EC 3.5.5.1) catalyzes the hydrolysis of indole-3-acetonitrile (IAN) to indole-3-acetic acid (IAA). Arabidopsis thaliana genome has four nitrilase genes (NIT1, NIT2, NIT3 and NIT4). Three (NIT1, NIT2 and NIT3) of the four genes have high similarity. We have cloned two NIT4 homologs (TNIT4A and TNIT4B) from tobacco (Nicotiana tabacum). Genomic Southern hybridization, among other experiments, strongly suggests that tobacco has NIT4 homologs but not NIT1 to NIT3 homologs. Introduction of Arabidopsis NIT2 into tobacco conferred IAN-mediated growth inhibition, probably due to hydrolysis of IAN to IAA, while ectopic expression of TNIT4A had little effect on the sensitivity of transgenic plants to IAN. Nitrilase activity of TNIT4 proteins is discussed.

A case-control study evaluating occult blood screening for colorectal cancer with hemoccult test and an immunochemical hemagglutination test.

A case control study to evaluate the occult blood screening for colorectal cancer was conducted in a town where colorectal cancer screening had been performed by Hemoccult test during the early years and subsequently by an immunochemical hemagglutination test. All residents aged >/=40 years had been offered the annual screening. Case series consisted of 51 subjects with fatal colorectal cancer. Three controls per case were selected from the list of residents who were alive at the time of diagnosis of the corresponding case and had been living in the town, matched by gender and by age. The odds ratio (OR) of dying of colorectal cancer for those having their most recent screening histories with Hemoccult test or the immunochemical test during the preceding 1 year and 1-2 year segment before case diagnosis were 0.20 [95% confidence interval (CI): 0.08-0.49] and 0. 17 (95% CI: 0.04-0.75), respectively. The OR increased towards 1.0 as the number of years since the most recent screening increased. The OR of dying of colorectal cancer was calculated to be 0.19 (95% CI: 0.05-0.70) for those screened with the immunochemical test alone during the preceding 1 year after adjustment for previous screening histories with the Hemoccult test. Corresponding OR was 0.36 (95% CI: 0.11-1.17) for those screened with Hemoccult test during the preceding 1 year. These results suggest that screening for colorectal cancer by fecal occult blood testings or immunochemical test alone would reduce mortality and that efficacy of the screening would be higher for the immunochemical test than for Hemoccult test.

Total number and size distribution of hepatic metastases of carcinoma.

OBJECTIVE: To quantify the susceptibility of carcinoma to hepatic metastases by studying autopsy livers with carcinoma metastases, the primary sites of which were mostly the digestive organs. STUDY DESIGN: We developed a stereologic method of estimating the total number, N, and the size distribution of metastatic tumors in the liver based on a geometric model of spherical nodules with varying radii, r. This method proved to be sufficiently reliable by disector analysis simultaneously performed in some cases; it gave an approximate result. This method was applied to the 31 autopsy cases. Correlation and regression analyses were performed among N, the mean radius of nodules, rmean, and conventional pathologic features of the primary tumor. RESULTS: The estimates of N ranged from 10 to 3.2 x 10(5). A close negative correlation was confirmed between N and rmean. Neither significant correlation nor regression was observed among N and the other pathologic factors of the primary tumors. CONCLUSION: N turned out to serve as a useful index for evaluating the metastatic potential of a carcinoma. However, investigation has yet to be made to determine biologic factors in the primary tumor closely associated with N.

Effects of Nd:YAG laser irradiation on morphometry and lung function in elastase-induced emphysema in rats.

BACKGROUND & OBJECTIVE: Although thoracoscopic laser ablation therapy has been hailed as an effective surgical treatment for diffuse emphysema, no one has as yet made an in-depth study of the efficacy of this treatment. This investigation was undertaken to research the effects of laser pneumoplasty on an animal model of emphysema. STUDY DESIGN/MATERIALS AND METHODS: Eight weeks after elastase treatment, the rats' left lungs were irradiated using contact Nd:YAG laser. Pulmonary function tests were performed 4 weeks after irradiation and the lungs were prepared for histologic examination. RESULTS: Dense fibrous scars beneath the pleura were observed at 4 weeks after irradiation. Although mean linear intercept values of irradiated lungs were not much lower than those in the non-irradiated elastase-treated group, laser irradiation caused a significant decrease in lung volume. While there was no significant difference in quasistatic compliance, elastic recoil pressure of the lung increased to control levels at total lung capacity volume. CONCLUSION: We conclude that laser therapy does not cause normalization of compliance, or improvement in the deeper part of the emphysematous lung, but rather a peripheral volume reduction and "encasement effect" on the lungs as a result of fibrotic scars.

[Histological effects of short term endocrine therapy on prostatic cancer].

BACKGROUND: The objective of this study is to investigate the pathological changes which occurred in prostatic cancer shortly after the commencement of endocrine therapy. METHODS: Fourty-three patients underwent radical prostatectomy immediately after the short term endocrine therapy (treatment period was within one month) and the histological pictures of operative specimens were compared to those obtained from the pretreatment biopsy specimens. RESULTS: Degenerative changes of cancer cells, such as nuclear and cytoplasmic vacuole, collapse of the cytoplasm and the appearance of naked hyperchromatic nucleus were noticed after the short term endocrine therapy. Especially in the cases which were histologically evaluated to be poorly differentiated in the biopsy specimens, not only degenerative changes but also destruction of cancer nests caused by cell death were observed. The histological effects affected by short term endocrine treatment had no relation to the prognosis, but in the cases of stage D2, the pathological grade judged by post-therapeutic specimens were found to be useful for the prediction of prognosis. CONCLUSION: Endocrine therapy induces remarkable pathological changes in prostatic cancer within a very short time after beginning treatment.

Papillary adenoma of type II pneumocytes might have malignant potential.

Papillary adenoma of type II pneumocytes is a rare tumour. It is considered to be a benign neoplasm and is derived from immature cells in the bronchioloalveolar epithelium, however, its biological nature has not been elucidated. We report a case of an adenomatous tumour; a papillary adenoma of type II pneumocytes, which we regard as possessing malignant potential. Light microscopically, as well circumscribed, papillary tumour of predominantly cuboidal cells resembling type II pneumocytes was found, but Clara type and ciliated cells were also present. Immunohistochemically, the tumour cells reacted positively with antibodies to surfactant apoproteins (A, B), carcinoembryonic antigen, cytochrome P-450 1A1-2 and 2B1-2. Ultrastructurally, many osmiophilic lamellar bodies and electron-dense granules were demonstrated. Semi-serial sections revealed signs of transbronchial dissemination and vascular invasion. Morphometry using 12-dimensional cluster analysis disclosed features of the tumour cells which resembled those of pneumocyte type II adenocarcinoma. These findings suggest that the present case has some malignant characteristics and originates from immature bronchiolar or alveolar cells, with a potential to develop into both type II pneumocyte and Clara cell type adenocarcinomas.

Atypical adenomatous hyperplasia and adenocarcinoma of the human lung: their heterology in form and analogy in immunohistochemical characteristics.

BACKGROUND: In a previous study, morphometry and multivariate cluster analysis was performed on 97 lesions. These consisted of atypical adenomatous hyperplasia (AAH), considered to be an important lesion corresponding to a step of carcinogenesis for adenocarcinoma of the human lung, Clara cell type, and type 2 pneumocyte type adenocarcinomas. Although AAH and the two types of adenocarcinoma were re-classified into three clusters and AAH was defined in clear morphologic terms, the biologic nature of AAH has yet to be clarified. In the present study, immunohistochemical analysis was performed to gain a deeper understanding of the relationship between AAH and the two types of adenocarcinoma, and to compare the results with those obtained by morphometry. METHODS: The 97 lesions analyzed by morphometry were submitted to immunohistochemical analyses using antibodies against surfactant apoprotein A, urine protein 1, carcinoembryonic antigen and cytochrome P-450s (1A1-2, 2B1-2, 2E1). Also examined, as controls, were 17 lesions with adenomatous hyperplasia (AH), a non-neoplastic reactive change of bronchiolo-alveolar cells, 30 areas of normal Clara cells, and 36 areas of normal type 2 pneumocytes. The immunoreactivity was graded by introducing a semi-quantitative scoring system. RESULTS: Immunohistochemically, AAHs behaved quite similarly to the lesions classified as Clara cell type or type 2 pneumocyte type adenocarcinomas. For any of the antibodies employed, no significant difference in immunoreactivity was demonstrated among these lesions. CONCLUSIONS: The results suggest, in accordance with our previous morphometry, that AAH is a lesion closely related with Clara cell type and type 2 pneumocyte type adenocarcinomas, probably as their common precursor. However, the two types of adenocarcinomas, despite their characteristic morphologic features, are indistinguishable using the biological indicators applied in this study.

Computer-assisted pathology of intraepithelial adenocarcinoma and related lesions: 3-D distribution, structural aberration and discrimination.

To discriminate among intraepithelial neoplasms, we have been relying on tissue microscopy, but pathologists' subjectivity sometimes impairs diagnosis. Even an individual pathologist is sometimes unable to reproduce exactly his or her own previous diagnosis. Are various atypical lesions classifiable in a reproducible way, and if they are, how? The reliability of a diagnosis will be strengthened if we can define the "natural" categories inherent in cells and tissues. Morphometry and statistical analysis using a computer can provide answers. Atypia, a morphological feature of carcinoma, is essentially multivariate. Quantification of a tissue feature requires reducing it to a set of ten or more quantities, including size, shape and position of the nucleus, nucleolus, and the cell itself. The grade of aberration from the norm can be assessed only by a synthetic approach, using a computer for multivariate cluster analysis. This classification has been attempted in adenocarcinoma and related lesions of the lung and pancreas. The categories thus established are reproducible, because the lesions fall into divisions according to their forms. We can also examine the organ distribution of intraepithelial neoplasms by three dimensional (3-D) computer-assisted mapping. To reach a higher level of reliability, as many meaningful features as possible should be taken into account. Particularly, we emphasize the significance of architectural pattern as a biomarker for intraepithelial glandular neoplasms. Computer-aided 3-D structural analysis visualizes the basic skeleton of these neoplasms around which the cells adhere. Instead of the dichotomous tree pattern of normal glands, the tumors basically harbor a 3-D network, tubular or porous, which increasingly deviates from the norm along with the transition from adenoma to well to moderately to poorly differentiated adenocarcinoma. This structural aberration, if recognizable on 2-D sectional images, will serve as a surrogate endpoint biomarker for glandular tumors.

Quantitative cytopathology of endometrial lesions.

Morphometric and multivariate statistical methods were used to discriminate endometrial carcinoma from benign cells in cytologic studies. Clumps of epithelial cells that appeared most diagnostically relevant were selected from aspirated samples of 70 endometrial cancer patients. The cells' cytologic character was reduced to a combination of five quantitative parameters--nuclear size, degree of anisokaryosis, nuclear from index, homogeneity of nuclear chromatin texture, and regularity of nuclear arrangement. The 5-variate cluster analysis demonstrated that the 70 cases could be classified into three definite groups: Group A (17 cases) was characterized by cells of small nuclear size, slight anisokaryosis, homogeneous chromatin texture, and regular nuclear arrangement; Group C (12 cases) by cells of large nuclear size, marked anisokaryosis, heterogeneous chromatin texture, and irregular nuclear arrangement; and Group B (41 cases) by cells of intermediate parameter values. Group C was derived from 10 cases of adenocarcinoma and 2 of atypical hyperplasia, while Groups A and B were not derived from any cases of malignancy. The computer-assisted morphometric statistical method can objectively classify the endometrial cells into malignant and benign, with improved validity and reproducibility. The cytopathologic finding, if detected by this method, may serve as a surrogate endpoint biomarker.

Morphometric and multivariate statistical detection of cancer cells in endometrial cytology.

Endometrial carcinoma was discriminated in cytologic studies using morphometric and multivariate statistical methods. On aspirated samples from 70 cases (10 well-differentiated adenocarcinomas, 4 hyperplasias and 56 normal controls), clumps of epithelial cells that could be regarded as the most diagnostically relevant were selected in each case. Their cytologic character was reduced to a combination of five quantitative parameters (nuclear size, degree of anisokaryosis, nuclear form index, homogeneity of nuclear chromatin texture and regularity of nuclear arrangement). The five-variate cluster analysis demonstrated that the 70 cases could be classified into three groups: group A (17 cases) was characterized by cells with small nuclear size, slight anisokaryosis, homogeneous chromatin texture and regular nuclear arrangement; group C (12) by cells with large nuclear size, marked anisokaryosis, heterogeneous chromatin texture and irregular nuclear arrangement; and group B (41) by cells with intermediate parameter values. Group C was derived from 10 cases of adenocarcinoma and 2 of atypical hyperplasia, while groups A and B were not derived from any cases of malignancy. This preliminary study indicated that morphometric-statistical classification can be of great help in improving the cytodiagnostic validity and reproducibility of endometrial carcinoma, and it awaits further, practical testing within a significantly larger series of malignancy cases.

Varying grades of epithelial atypia in the pancreatic ducts of humans. Classification based on morphometry and multivariate analysis and correlated with positive reactions of carcinoembryonic antigen.

To establish an adequate statistical classification of epithelial atypia in the pancreatic duct, a total of 78 areas of duct epithelia with varying grades of atypia were subjected to nine-parameter morphometry and nine-dimensional multivariate cluster analysis. The material was derived from 53 pancreases resected for various epithelial tumors or acute or chronic pancreatitis. The result was correlated with immunohistochemical findings in which the pattern of intraepithelial distribution of carcinoembryonic antigen changed with the degree of ductal atypia. Finally, atypical cells classified by cluster analysis and immunohistochemistry were subjected to computer-aided three-dimensional mapping to visualize their distribution in the ductal tree. Cluster analysis demonstrated that the various epithelial forms were classifiable into Clusters 1, 2, and 3, representing ordinary epithelia and mild and severe dysplasias, respectively. The last category was created so as to include not only in situ and invasive carcinoma but the so-called borderline atypical lesions. The reproducibility of this classification was proved by two sorts of discriminant analyses. Also, the grades of atypia shown by the clustering proved to correlate with the reaction patterns for carcinoembryonic antigen. In the computer-aided three-dimensional mapping, severely dysplastic areas were shown surrounded by zones of mild dysplasia, justifying the assumption of a stepwise carcinogenesis in the pancreatic ducts.

[Flow cytometric analysis of prostatic adenocarcinomas after hormone therapy].

A DNA ploidy pattern of prostatic adenocarcinoma was studied by flow cytometry using the formalin-fixed, paraffin-embedded tissues obtained from 65 patients. These patients received hormone therapy preoperatively. Forty cases (61.5%) showed a diploid pattern, and 25 (38.5%) an aneuploid pattern. Comparing this ploidy pattern with histological classification, the aneuploid pattern was found in 13 cases (52%) of poorly differentiated group, 12 cases (48.0%) of moderately differentiated group and 0 case (0%) of well differentiated group. Thus, rate of aneuploid pattern increased with an increase in the grade of malignancy. Five years survival rate was 62.6% in the diploid group while it decreased to 30.9% in the aneuploid group. There was a significant difference between the two groups. In 23 cases with stage D, the ploidy pattern of primary region could be compared with that of metastatic region (lymph nodes). The prognosis of patients who exhibited the aneuploid pattern in metastatic region was significantly worse than that of other patients. These data suggested that flow cytometric DNA analysis of prostatic cancer materials after hormone therapy also produces meaningful information in predicting the prognosis of prostatic cancer.

[Morphometrical and statistical detection of cancer cells].

Morphometrical and multivariate statistical methods were applied to detection of carcinoma cells in cytologic studies. A total of 70 clumps of epithelial cells were selected from endometrial samples (10 adenocarcinomas, 4 hyperplasias and 56 normal controls), and their cytologic characters were reduced to a combination of five quantitative parameters (nuclear size, degree of anisokaryosis, nuclear from index, homogeneity of nuclear chromatin texture and regularity of cellular arrangement). The 5-variate cluster analysis demonstrated that the 70 clumps could be classified into three definite groups; A(17), B(41), and C(12). Group C, characterized by cells of large nuclear size, marked anisokaryosis, heterogeneous chromatin texture and irregular cellular arrangement, was derived from 10 samples of adenocarcinoma and 2 hyperplasia, while Groups A and B were not derived from any malignant samples. In conclusion, morphometrical-statistical classification can be of great aid in improving the cytodiagnostic validity and reproducibility.

[Two cases of primary sclerosing lipogranuloma in scrotum--review of 63 cases reported in Japan].

We reported two cases of primary sclerosing lipogranuloma in the scrotum. We performed tumor resection in both cases, but in one of the two cases tumor recurrence was observed 7 days after the removal. Sixty-three cases have been reported in our country, and we discuss the diagnosis and treatment with reference to previous reports.

Atypical adenomatous hyperplasia of the lung and its differentiation from adenocarcinoma. Characterization of atypical cells by morphometry and multivariate cluster analysis.

BACKGROUND: Atypical adenomatous hyperplasia (AAH) of the human lung is considered to be an important lesion preceding adenocarcinoma, but its difference from well-differentiated adenocarcinomas is so subtle as to cause diagnostic uncertainty. In view of this, the authors undertook to establish reproducible microscopic criteria for AAH, Type II pneumocyte type, and Clara cell type adenocarcinomas. METHODS: Twelve-parameter morphometry of atypical cells was performed on 97 lesions selected from 303 surgical specimens of lung by routine microscopic examination: all were considered by premorphometry examination to correspond to one of the above three diseases. Measurements were performed on photomicrographs using a digital image analyzer. The data of morphometry were subjected to 12-variate cluster analysis using a mainframe computer. RESULTS: It was demonstrated that the lesions were classifiable into three groups: Cluster 1 (Type II cell adenocarcinoma and AAH), Cluster 2 (AAH), and Cluster 3 (Clara cell adenocarcinoma). Whereas the latter two were created as homogeneous clusters, Cluster 1 was a mixture of Type II tumors and AAH. CONCLUSIONS: AAH in the strict sense of the word is definable by the features of those classified into Cluster 2, with atypia milder than overt adenocarcinomas. These AAH are likely to correspond to one of the steps of carcinogenesis forgoing the final one. The lesions, diagnosed as AAH before morphometry and being assigned to Cluster 1 and therefore not separable from Type II carcinoma, are considered to have been Type II carcinoma from the very beginning, which were however underdiagnosed in routine microscopic examination.

Development of the human colonic adenocarcinoma from adenoma as a histopathologically continuous process.

The adenocarcinoma sequence of the human colon is usually divided into three separate steps according to the grade of epithelial dysplasia, i.e., adenomas with mild, moderate and severe dysplasia. In an attempt to re-examine whether or not this sequence can be morphologically separable as usually assumed, a total of 192 epithelial lesions including adenomas with various grades of dysplasia were analyzed, relying on morphometrical and multivariate-statistical techniques. Histologic features of epithelial lesions were characterized by 10 quantitative parameters and subjected to 10-variate cluster analysis. In the result of computations, separation of neoplastic lesions into different groups proved to be rather ambiguous, not justifying the classification into three groups as generally expected. It was concluded that adenocarcinoma can develop from adenoma as a seemingly continuous process which may involve more steps than usually assumed.

[The grading of cellular and structural atypia in cancer and related lesions using multivariate analysis].

In morphologically diagnosing cancer, pathologists' attention is focussed on the presence or absence of atypia, i.e., the form of cells and tissues deviated from the norm, and if it is present, on its grade. However, due to retarded development of techniques for the objective evaluation of its grade, separation of cancer from premalignant lesions still meets great ambiguity. Particularly, the boundaries of "dysplasia" with cancer and noncancerous lesions are not defined in clear morphological terms and are susceptible to a strong between-observers fluctuation. In this paper we outlined our recent efforts to establish in various organs a statistically most adequate, and therefore reproducible, classification of atypical lesions, resorting to morphometry and multivariate analysis. Two examples were given: atypia of the pancreatic duct epithelia as an object for the study of purely cellular abnormalities, and hepatocellular carcinoma, which required also to quantify the atypia of tissue structures, The classifications thus established proved to be the most adequate, in that, they closely reflected the grade of malignancy in a clinical as well as biological sense, as shown by the clear between-categorier differences in oncogene mutation, DNA ploidy pattern, the patients' prognoses, and so on.