Ovarian teratoma displaying a wide variety of tissue components in a broiler chicken (Gallus Domesticus): morphological heterogeneity of pluripotential germ cell during tumorigenesis.

Spontaneous ovarian teratoma was found in a seven-week-old female Chunky broiler chicken that was slaughtered for food. On post-mortem inspection, a spherical tumor mass attaching to a juvenile ovary was found in the abdominal cavity. Histopathologically, the tumor was comprised of immature mesenchymal stroma and a variety of mature tissue elements of mesodermal and ectodermal origin. In addition, there were multiple indistinguishable tissue elements, which showed no malignant cytological features but were unidentifiable as to corresponding embryological layer of origin. These heterogeneous teratoma tissues consisted of a variety of glandular, cystic, duct-like, and tubular structures, some of which exhibited a lining by a mixture of both keratinizing/non-keratinizing stratified squamous epithelial cells and cuboidal/columnar epithelial cells. The ovarian tetatoma was considered a benign and congenital one. The highly diverse differentiation of the teratoma might have manifested a morphological aspect of intrinsic character of the pluripotential germ cells during tumorigenesis.

Emphysematous Eosinophilic Lymphangitis in the Ruminal Submucosa of Cattle.

Twenty cattle (14 Holstein-Friesian, 3 Japanese Black, 3 Aberdeen Angus) ranging in age from 3 months to 8 years exhibited, at slaughter, emphysematous thickening of the ruminal submucosa owing to the appearance of numerous, contiguous, small gas bubbles. Microscopic changes in the ruminal submucosa consisted of (1) multiple cystic (emphysematous) lymphangiectasis that was frequently lined or occluded by granulomatous inflammatory infiltrates including macrophages, multinucleate giant cells, and eosinophils; (2) intralymphatic phagocytosis by macrophages and giant cells of eosinophils that showed positive labeling with the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling assay; and (3) an inflammatory infiltrate extending from the area of lymphangitis into surrounding tissue, as well as edema, hemorrhage, fibrin exudation, fibroplasia, or capillary proliferation throughout the lesional submucosa. In addition, 15 (75%) of the cattle had globular leukocyte infiltrates in the mucosal epithelia of the rumen.

Epidermoid cysts of the rumen in a Holstein-Friesian steer.

Epidermoid cysts were identified in the rumen of a 2-year-old Holstein-Friesian steer. Small nodules (<4 mm diameter) filled with paste-like, friable, white material were scattered in the rumen wall. Microscopically, the nodules were located in the submucosal connective tissue and comprised of cysts lined by stratified squamous epithelium surrounding keratin debris, exhibiting features identical to those of epidermoid cysts. The cysts were surrounded by chronic inflammation. The ruminal mucosa contained vesicles or cystic cavities filled by inflammatory exudate, in addition to empty cystic spaces lined by flattened endothelial cells, which were possibly of lymphatic origin. The development of these epidermoid cysts might have resulted from implantation of exfoliated squamous epithelial cells from damaged ruminal epithelium into the submucosal connective tissue, although the precise aetiology of this anomaly remains to be defined.

Maternal fat consumption during pregnancy and risk of wheeze and eczema in Japanese infants aged 16-24 months: the Osaka Maternal and Child Health Study.

BACKGROUND: One factor capable of modulating antenatal immune responses is diet. This prospective study examined the association between maternal intake of specific types of fatty acids, cholesterol, fish and meat during pregnancy and the risk of wheeze and eczema in the offspring. METHODS: Subjects were 763 mother-child pairs. Data on maternal intake during pregnancy were assessed with a diet history questionnaire. Data on wheeze and eczema based on criteria of the International Study of Asthma and Allergies in Childhood were obtained from a questionnaire completed by mothers 16-24 months postpartum. RESULTS: Higher maternal intake of alpha-linolenic acid and docosahexaenoic acid during pregnancy was independently associated with a reduced risk of wheeze, but not eczema, in the offspring (adjusted odds ratios (ORs) between extreme quartiles 0.52 (95% CI 0.28 to 0.97) and 0.37 (95% CI 0.15 to 0.91), respectively). Higher maternal intake of n-6 polyunsaturated fatty acids and linoleic acid during pregnancy was independently related to an increased risk of infantile eczema but not wheeze (adjusted ORs 2.25 (95% CI 1.13 to 4.54) and 2.11 (95% CI 1.06 to 4.26), respectively). No significant exposure-response relationships were observed between maternal consumption of total fat, saturated fatty acids, monounsaturated fatty acids, eicosapentaenoic acid, arachidonic acid, cholesterol, meat and fish and the ratio of n-3 to n-6 polyunsaturated fatty acid consumption during pregnancy and infantile wheeze or eczema. CONCLUSIONS: Maternal intake of alpha-linolenic acid and docosahexaenoic acid during pregnancy may be preventive against infantile wheeze. Maternal intake of n-6 polyunsaturated fatty acids, especially linoleic acid, during pregnancy may increase the risk of childhood eczema.

Diffuse coronary arteriodysplasia in a Holstein-Friesian cow.

The coronary arteries of a 6-year-old Holstein-Friesian cow showed dysplastic lesions characterized by the following vascular changes: irregular, aneurysmal dilation; intimal fibrosis associated with myointimal cells; aberrant internal elastic lamina; oedema and mast cell infiltrates; and variation in the thickness of the media, including tortuosity with irregular dilation of the lumen of the artery and a plexiform structure formed from the right coronary artery.

Spinal dysraphism in a newborn Holstein-Friesian calf.

Spinal dysraphism, not associated with vertebral defect or arthrogryposis, was found in a 3-day-old Holstein-Friesian calf that was clinically diagnosed as having encephalopathy. The dysraphic lesion occurred in the sixth (C6) and seventh (C7) segments of the cervical spinal cord. Microscopically, the lesion was characterized by hydromyelia, syringomyelia, anomaly of the ventral median fissure, abnormal running of the myelinated nerve fibers in the white column, and absence of the central canal due to a developmental defect of the ependymal cells.

Pretreatment with restriction enzyme or bovine serum albumin for effective PCR amplification of Epstein-Barr virus DNA in DNA extracted from paraffin-embedded gastric carcinoma tissue.

An association between Epstein-Barr virus (EBV) and gastric carcinoma has been studied through the EBV genome present in the carcinoma cells. Recently, we found that EBV DNA in paraffin-embedded gastric carcinoma tissue was detected effectively by PCR after pretreatment of the extracted DNA with a restriction enzyme, BamHI or EcoRI. Here, we show that the PCR amplification was also enhanced by pretreatment of the DNA with other restriction enzymes or with bovine serum albumin and several other proteins. Treatment with these proteins may remove a PCR inhibitor(s) in the DNA samples extracted from the paraffin blocks.

Evidence that expression of a mutated p53 gene attenuates apoptotic cell death in human gastric intestinal-type carcinomas in vivo.

To examine in vivo the validity of the results of experiments in vitro, we analyzed the relationship between p53 gene status and apoptotic cell death of human gastric intestinal-type adenocarcinomas. Surgical specimens were classified into two categories: 18 gastric cancers with nuclear p53 protein (A), and 17 gastric cancers without nuclear p53 protein (B). Polymerase chain reaction-single strand conformation polymorphism disclosed a shifted band that corresponded to a mutation in the p53 gene in 13 cases (72%) in category A and 3 cases (18%) in category B, the frequency being significantly higher in the former (P < 0.05). Apoptotic cells were identified from routinely stained sections and by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL). The TUNEL index [TI; (the number of TUNEL-positive apoptotic cells/the total number of tumor cells) x 100] was 3.8 +/- 1.4% in category A and 4.9 +/- 1.2% in category B, the value being significantly lower in the former (P < 0.05). The proliferating cell nuclear antigen index, defined similarly to the TI, was 56.4 +/- 16.3% in category A, and it was significantly higher than that in category B (P < 0.05). The immunohistochemically detected expression of p21CIP1/WAP1 did not differ between the two categories, while Bax-positive tumor cells were more frequently detected in category A. These results indicate that (1) expression of a mutated p53 gene attenuates apoptotic cell death of gastric cancer, in accordance with the previous in vitro finding that p53 gene mutation provides a possible selective advantage for tumor cell proliferation, and (2) apoptosis is related not only to expression of p53 and the stage of the cell cycle, but also to p53-independent and cell cycle-independent events.

Low frequency of apoptosis in Epstein-Barr virus-associated gastric carcinoma with lymphoid stroma.

In order to understand the character of Epstein-Barr virus (EBV)-associated gastric carcinoma with lymphoid stroma (GCLS), we examined cell proliferation and death in comparison with gastric carcinoma without prominent lymphoid stroma (conventional gastric carcinoma). The EBV-encoded small RNA I (EBER-1) and apoptotic cells were examined by in situ hybridization (ISH) and by terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick end-labeling (TUNEL), respectively, on formalin-fixed paraffin-embedded surgical specimens from 22 GCLSs and 23 conventional gastric carcinomas. Immunostaining was performed for the detection of Ki-67 antigen (Ki-67), P53 protein (P53) and bcl-2 oncoprotein (BCL-2). The percentages of apoptotic and Ki-67-positive cells were expressed as apoptotic indices (AI) and Ki-67 labeling indices (KI), respectively. EBER-1 was detected in 19 (86%) GCLSs in contrast to none in conventional carcinomas. Mean AI was 1.8 +/- 0.6 in the EBER-1-positive GCLSs and 3.3 +/- 1.7 in the conventional gastric carcinomas, the value being significantly lower in the former (p < 0.01). Mean KI was 40.0 +/- 10.8 in the GCLSs and 48.5 +/- 9.1 in the conventional gastric carcinomas, the value being significantly lower in the former (p < 0.05). P53 expression showed no significant difference between the 2 categories of carcinoma.

Apoptosis in human gastric mucosa, chronic gastritis, dysplasia and carcinoma: analysis by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labelling.

We examined the existence and distribution of apoptotic cells in human gastric mucosa, chronic gastritis, adenomatous dysplasias and carcinomas in 15 surgically removed stomachs in which dysplasia and carcinoma were found simultaneously. Serial sections were cut for immunohistochemistry for p53 oncoprotein and Ki-67 antigen, and terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick end labelling (TUNEL). TUNEL signal-positive apoptotic cells were rare in normal mucosa, while a few apoptotic cells were noted in gastritic mucosa and intestinal metaplasia, intermingled with Ki-67 antigen-positive cells forming a generative cell zone. This suggests the cell-cycle-dependent apoptosis of gastric mucosa. The frequency of apoptotic cells per crypt was higher in incomplete than in complete metaplasia, implying greater underlying DNA damage in the former. TUNEL indices (TI: percentage of TUNEL-positive cells in tumour cells) were slightly higher in adenomatous dysplasias (4.9 +/- 2.1) than in carcinoma (3.9 +/- 1.1), but there was no no statistical difference. Ki-67 indices (KI: percentage of Ki-67 antigen-positive cells in tumour cells) were significantly (P < 0.05) higher in carcinomas than in dysplasias. Thus, gastric adenomatous dysplasias were characterized by relatively higher TI and lower KI, which might reflect a more static growth potential. The expression of p53 oncoprotein in cancer cells is thought to be an apoptosis-suppressing event, although its precise role remains to be elucidated. Overall, these results indicate that apoptosis plays a crucial part in the morphogenesis of gastritic mucosa including intestinal metaplasia, and that the process is correlated both with tumourigenesis and with proliferative activity.

Apoptotic cell death and its relationship to carcinogenesis in colorectal carcinoma.

BACKGROUND: Apoptotic cell death plays an important role in the proliferation and turnover of cells in various tumors. The relationship between apoptosis and cell proliferation was studied to determine each of their roles in colorectal carcinogenesis. METHODS: Apoptotic cells were identified by the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) method. The occurrence of apoptosis was examined in colorectal cancer that had invaded the submucosa. Specimens were obtained from 38 cases of cancer with adenoma and 29 cases of cancer de novo. Apoptotic indices (AIs) as percentages of TUNEL-positive cells relative to the number of tumor cells and Ki-67 labeling indices (PI) were investigated. The relationship between the frequency of apoptosis and the expression of p53 and c-myc proteins was also investigated. RESULTS: In cancer with adenoma, the ratio of AI/PI in adenoma cells was significantly higher than that of cancer cells (P < 0.0001). Mean AIs of cancer with adenoma were significantly higher than those of cancer de novo particularly in the flat-type cancers (P < 0.05). Among p53 negative tumors, the ratio of AI/PI for cancer de novo was significantly lower than that for cancer with adenoma (P < 0.05). AI of cancer de novo was lower than that of cancer with adenoma in cases with overexpression of c-myc protein (P < 0.05), whereas there was no significant difference in the ratio of AI/PI between cancer de novo and cancer with adenoma. CONCLUSIONS: Colorectal carcinogenesis is related to the inhibition of apoptosis and to the augmentation of proliferative activity both in cancer with adenoma and in cancer do novo. A reduction of the rate of apoptosis as compared with that of cell proliferation might explain the rapid-growing nature of cancer de novo particularly in cases with the flat-type appearance.

Synchronous and metachronous primary malignancies in organs other than the stomach in patients with early gastric cancer.

BACKGROUND/AIMS: We investigated the outcomes of patients with early gastric cancer, with special reference to the prognosis of patients with synchronous or metachronous primary malignancies in organs other than the stomach. PATIENTS AND METHODS: Among 890 patients with early gastric cancer, 97 (10.9%) had synchronous or metachronous primary malignancies in organs other than the stomach. Ten-year survival rates were compared between patients who had additional malignancies and patients who had early gastric cancer but no other malignant disease (control group). RESULTS: Synchronous primary malignancies were detected in 32 patients and metachronous primary malignancies were detected in 65 patients (17 had developed before gastrectomy and 48 developed after gastrectomy). Hepatic cell carcinoma, lung cancer and colorectal cancer were frequently detected between 2 and 24 years after gastrectomy. The 10-year survival rate was 80.8% for 769 patients in the control group but it was only 49.7% for the 92 patients with additional malignancies. Moreover, metachronous malignant disease was found more over 10 years after gastrectomy in 30 of the 48 cases (62.5%). CONCLUSIONS: These results suggest the importance of long-term follow-up for detection of metachronous carcinomas at sites other than the stomach for patients with early gastric cancer.

Aneuploidy of tumor cells in cases of gastric cancer with esophageal invasion: another indicator of poor prognosis.

The prognosis of patients with gastric cancer with esophageal invasion is extremely poor. To evaluate factors related to this poor prognosis, we analyzed 200 patients with gastric cancer located in the upper third of the stomach. These patients underwent gastrectomy and were divided into two groups in terms of the presence (E[+] group; n = 62) or absence (E[-] group; n = 138) of histological evidence of esophageal invasion. Even when apparently curative surgery was performed, the 5-year survival rate of patients with E[+] gastric cancer (45.8%) was significantly lower than of patients with E[-] gastric cancer (71.6%). In the E[+] group, the 5-year survival rate of patients who had tumors with infiltrative growth and DNA aneuploidy was only 10.0%. These patients had a high frequency of peritoneal metastasis at operation (5/16; 31.3%); even when apparently curative operations were performed, 50% of these patients died from peritoneal metastatic recurrence within 2 years after surgery. Gastric adenocarcinoma with esophageal invasion accompanied by infiltrative growth and DNA aneuploidy had a high potential for peritoneal metastasis. This combination is associated with the most pessimistic prognosis for patients with gastric cancer with esophageal invasion.

Hepatocholangioadenoma in a pig.

A Landrace sow had several neoplastic nodules in the liver. Histopathologically, the individual nodules consisted of two types of closely associated neoplastic cells; one was of hepatocellular type, the other of cholangiocellular type. Based on this finding, the term "hepatocholangioadenoma" is considered to be appropriate for this tumour. Previously reported in man, the dog and the duck, a mixed type of primary hepatic neoplasm has now been found in the pig.

Lactate dehydrogenase isoenzyme patterns in bovine liver tissue.

Liver lactate dehydrogenase (LDH) isoenzymes in normal and diseased cows were analyzed electrophoretically. This method (histoelectrophoresis) was improved for the direct analysis of tissue LDH isoenzymes. The mean values of LDH 1 through LDH 5 in the livers of normal cows were 31.7, 24.8, 27.3, 12.8, and 3.3%, respectively. In cases with hydropic degeneration of the liver, the patterns revealed increases of LDH 1 and LDH 2 as compared to normal cows. The patterns showed a decrease of LDH 1 and an increase of LDH 2 in fatty change of the liver. Congestion of the liver alone decreased LDH 1 and increased LDH 3, LDH 4 and LDH 5. Necrosis of the liver decreased LDH 1 and LDH 2, and increased LDH 3, LDH 4 and LDH 5. It was suggested that the functional hepatocellular damage due to anoxia might be a important factor of the change of liver LDH isoenzyme patterns. We have attempted to standardize the LDH isoenzyme patterns by using a computer under various conditions. In cases with hydropic lesions, the diagnostic sensitivity (DS) was 53% (28 of 53 cases) and the predictive value of positive tests (PVPT) was 100% (28 cases of 28 cases selected). In cases with congestive or necrotic lesions, DS was 41% (9 of 22 cases) and PVPT was 69% (9 cases of 13 cases selected).