Touichiro Takizawa

Quantitative Analysis of Mycobacterial and Propionibacterial DNA in Lymph Nodes of Japanese and European Patients with Sarcoidosis

ABSTRACT The cause(s) of sarcoidosis is unknown. Mycobacterium spp. are suspected in Europe and Propionibacterium spp. are suspected in Japan. The present international collaboration evaluated the possible etiological links between sarcoidosis and the suspected bacterial species. Formalin-fixed and paraffin-embedded sections of biopsy samples of lymph nodes, one from each of 108 patients with sarcoidosis and 65 patients with tuberculosis, together with 86 control samples, were collected from two institutes in Japan and three institutes in Italy, Germany, and England. Genomes of Propionibacterium acnes, Propionibacterium granulosum, Mycobacterium tuberculosis, Mycobacterium avium subsp. paratuberculosis, and Escherichia coli (as the control) were counted by quantitative real-time PCR. Either P. acnes or P. granulosum was found in all but two of the sarcoid samples. M. avium subsp. paratuberculosis was found in no sarcoid sample. M. tuberculosis was found in 0 to 9% of the sarcoid samples but in 65 to 100% of the tuberculosis samples. In sarcoid lymph nodes, the total numbers of genomes of P. acnes or P. granulosum were far more than those of M. tuberculosis. P. acnes or P. granulosum was found in 0 to 60% of the tuberculosis and control samples, but the total numbers of genomes of P. acnes or P. granulosum in such samples were less than those in sarcoid samples. Propionibacterium spp. are more likely than Mycobacteria spp. to be involved in the etiology of sarcoidosis, not only in Japanese but also in European patients with sarcoidosis.

In situ localization of Propionibacterium acnes DNA in lymph nodes from sarcoidosis patients by signal amplification with catalysed reporter deposition

Sarcoidosis is a systemic granulomatous disease of unknown aetiology. Many genomes of Propionibacterium acnes and P. granulosum have been detected in lymph nodes from patients with sarcoidosis. In situ localization of propionibacterial genomes in sarcoid lymph nodes may help to establish an aetiological link between sarcoidosis and these indigenous bacteria. Formalin‐fixed and paraffin‐embedded biopsy samples of lymph nodes from nine patients with sarcoidosis, nine patients with tuberculosis, and nine patients with non‐specific lymphadenitis as controls were examined by quantitative real‐time PCR (QPCR) for P. acnes and by in situ hybridization (ISH) that used catalysed reporter deposition (CARD) for signal amplification with digoxigenin‐labelled oligonucleotide probes that complemented 16S rRNA of P. acnes. The signals per 250 µm2 of tissue sections were counted from inside and outside the granulomas of sarcoidosis and tuberculosis and from control lymph nodes. The number of genomes by QPCR was examined for correlation with the mean signal count by ISH with CARD. In sarcoid samples, one or several signals were detected in the cytoplasm of some epithelioid cells in granulomas and of many mononuclear cells around granulomas. The mean signal counts were higher (p < 0.001) in granulomatous areas than in other areas of sarcoid lymph nodes. Even in their non‐granulomatous areas, counts were higher than in granulomatous areas (p = 0.0023) and non‐granulomatous areas (p < 0.001) of tuberculous lymph nodes and control lymph nodes (p = 0.0071). Correlation between the results by QPCR and ISH with CARD was significant (r = 0.86, p < 0.001). The accumulation of P. acnes genomes in and around sarcoid granulomas suggests that this indigenous bacterium may be related to the cause of granulomatous inflammation in sarcoidosis. Copyright

Absence of either gastric or intestinal phenotype in microscopic differentiated gastric carcinomas

Differentiated gastric carcinoma (DGC) corresponds roughly to the intestinal type of gastric carcinoma described by Laurén. It has been suggested that DGCs arise from intestinalized gastric mucosa, but recent findings regarding their mucin expression do not support this hypothesis. To evaluate the histogenetic relationship between DGCs and intestinal metaplasia, lesions that are as small as possible should be examined. Twenty‐five DGCs, ranging in their greatest dimension from 0.4 to 2.7 mm, were collected and divided into two groups by size. Group A consisted of 13 lesions less than 1.4 mm across, and group B of 12 lesions 1.4 mm or more. The presence of mucin and a brush border was assessed by immunostaining with antibodies against human gastric mucin, pyloric‐gland‐type mucin, Muc‐2 glycoprotein, and CD10 antigen, and the lesions were classified as having the gastric phenotype (G‐type), intestinal phenotype (I‐type), mixed gastric and intestinal phenotype (M‐type), or null phenotype (N‐type). Thirteen (52%) of the 25 lesions were N‐type, 5 (20%) lesions were G‐type, 5 (20%) were I‐type, and 2 (8%) were M‐type. Group A had a larger proportion of N‐type lesions than B (10/13, or 77%, vs. 3/12, or 25%; p = 0.027, chi‐square test for proportions). Group B had a larger proportion of G‐type lesions than A (5/12, or 42%, vs. 0/13, or 0%; p = 0.033). The phenotypes of the carcinomas and their surrounding mucosa were unrelated. Therefore, DGCs may arise from gastric mucosa affected by intestinal metaplasia or not, without having either the gastric or intestinal phenotype. Copyright

AGE‐ASSOCIATED CHANGES IN MICROVASCULATURE OF HUMAN ADULT TESTIS

Age‐associated architectural changes of the human testicular micro‐vasculature from 70 autopsy cases were stereoscopically examined with a silicone‐rubber injection technique. In the testis of a young subject, the interlobular main arteries run straight. The coiling phenomena of the interlobular centripetal or centrifugal arteries, which are commonly seen in adult testis, have been so far considered as physiological transformation of the vasculature. It was confirmed that the coiling changes in the interlobular main arteries of the human testis appear as an age‐dependent alteration of the vasculature closely related to the volume of the gland. The practical importance of the spiralling or coiling of arteries is that it results in a considerable reduction of blood flow. The age‐related coiling of the interlobular arteries is virtually accompanied by varying degrees of collapse of the peritubular capillary networks. The reduction of blood supply to the seminiferous tubules plays an active role in promoting aging of the testis. These stereoscopical observations of age‐related transfiguration of testicular microvasculature were ascertained also by histometrical examinations.

Loss of uroplakin III expression is associated with a poor prognosis in patients with urothelial carcinoma of the upper urinary tract

To investigate the association between the expression of uroplakin III (UPIII) and the prognosis of patients with urothelial carcinoma of the upper urinary tract, as uroplakins are urothelium‐specific markers of terminal urothelial differentiation.

Schwannoma arising from the tentorium at an unusual location: case report.

OBJECTIVE AND IMPORTANCE We present a case of schwannoma attached to the tentorium. CLINICAL PRESENTATION A 41-year-old woman without evidence of neurofibromatosis presented with a 3-month history of headache, positional vertigo, and truncal ataxia. Magnetic resonance imaging revealed an extra-axial cystic mass lesion in the left anteromedial cerebellar region with a dural tail sign. INTERVENTION The tumor was removed completely by retrosigmoid craniotomy. Dense adhesion of the tumor to the inferior surface of the tentorium was confirmed during surgery. On light microscopic study, this neoplasm was composed of spindle cells and showed palisaded structures. Immunohistochemically, the tumor cells stained positive for S-100 protein and vimentin. Reticulin staining revealed a pericellular pattern of distribution of reticulin fibers. Electron microscopy confirmed the presence of a basement membrane encompassing the tumor cells. From these findings, the tumor was diagnosed as a schwannoma arising from the tentorium. CONCLUSION To our knowledge, this case report is the first to describe a schwannoma arising from the tentorium. Our case report indicates that schwannoma is a possible pathology in the differential diagnosis of dura-based tumors.

Basement Membrane Matrix Modifies Cytokine Interactions between Lung Cancer Cells and Fibroblasts

Objective: Proliferation of fibroblasts (desmoplastic reaction) in the lung adenocarcinomas is an important phenomenon that correlates with metastases and poor prognosis. Because basement membranes are often involved in the desmoplastic areas and many cytokines have binding capacity to basement membrane molecules, we hypothesized that basement membrane modify the paracrine effects between cancer cells and fibroblasts via the fibrogenic cytokines and this hypothesis was experimentally investigated. Methods: The effects of conditioned media derived from ten lung carcinoma cell lines and normal airway epithelial cells on DNA synthesis of fetal lung fibroblasts were determined. We focused on fibroblast growth factor 2 (FGF-2) as the candidate paracrine cytokines and examined their diffusion through an experimental basement membrane matrix model, Matrigel™. Results: All the conditioned media promoted DNA synthesis of fetal lung fibroblasts. Detection by ELISA methods and the neutralizing antibodies suggested that FGF-2 was one of the responsible factors for the growth promotion. Diffusion of FGF-2 across the polycarbonate membrane was suppressed by coating with Matrigel. When FGF-2-secreting A549 cells were covered with Matrigel, FGF-2 was stored in Matrigel and its diffusion into the culture media was significantly reduced. Binding of FGF-2 to Matrigel was completely blocked by a basic protein, protamine sulfate. In the presence of protamine sulfate in Matrigel overlaid on A549 cells, diffusion of FGF-2 increased 7-fold as much as that without overlaid Matrigel. Conclusion: These results suggest that the basement membrane acts as a barrier to the diffusion and a reservoir of cytokines secreted by cancer cells, and that the subsequent degradation of the basement membrane by cancer cells could release the stored cytokines and promote growth of fibroblasts.

Mucin core protein expression by colorectal mucinous carcinomas with or without mucus hyperplasia

BackgroundIn the categorization of colorectal mucinous carcinomas, much attention has been paid to the amount of extracellular mucin, but little to that of intracellular mucin. Perhaps this factor would be useful in further categorization.MethodsWe estimated the amount of intracellular mucin morphologically, and classified colorectal tumors (tubular adenomas, mucinous carcinomas, and nonmucinous carcinomas) into two categories each, those with and without intracellular mucus hyperplasia. From preliminary results, we chose a range of 50% or more for the ratio of intracellular mucus to the total area of tumor cells for our definition of such hyperplasia. Then, mucin expression in the different categories was examined immunochemically with antibodies to the mucin core proteins MUC1, MUC2, MUC5AC, and MUC6.ResultsMUC1 expression was found in none of the 40 adenomas, 8 (17%) of the 48 mucinous carcinomas (with little difference in those with and without mucus hyperplasia), and 4 (16%) of the 25 nonmucinous carcinomas. MUC2 was found in all mucinous carcinomas. MUC5AC was found in 18 (86%) of the 21 mucinous carcinomas with mucus hyperplasia and 18 (90%) of the 20 adenomas with mucus hyperplasia, but in only 9 (33%) of the 27 mucinous carcinomas without mucus hyperplasia, 5 (20%) of the 25 nonmucinous carcinomas, and 2 (10%) of the 20 adenomas without mucus hyperplasia.ConclusionsMucinous carcinomas with and without mucus hyperplasia may arise from adenomas with and without mucus hyperplasia, respectively. The amount of intracellular mucin may be a morphologic reflection of the origin of colorectal mucinous carcinomas.

Correlation between the expression of telomerase reverse transcriptase and proliferative activity in breast cancer cells using an immunocytochemical restaining method

Telomerase activity is thought to contribute to the immortality of cancers. Recently, some investigators described a correlation between the activity of telomerase and the proliferative activity of cancer cells. The aim of this study was to evaluate the correlation between the expression of telomerase‐associated protein and proliferative activity. Telomerase reverse transcriptase (TERT) is one of the proteins that correlates with telomerase activity. We investigated TERT protein and its mRNA, and examined the correlation between the TERT protein and Ki‐67, which reflects proliferative activity with immunostaining, and its mRNA, which correlates with telomerase activity, using in situ hybridization. Imprint smears from 17 invasive ductal adenocarcinomas were investigated. In most cases positive for TERT mRNA, the percentage of TERT protein‐positive cells was also high and was closely related to mRNA (P = 0.024). The positive rates of TERT for the cases with lymph node metastasis were significantly higher than those for the cases without metastasis (P = 0.046). The positivity of TERT protein also correlated significantly with the Ki‐67‐positive rate (r = 0.82). As the proliferation activity increased, the number of cells positive for both proteins also increased (r = 0.89). In conclusion, it was suggested that the expression of TERT  protein is associated with the expression of Ki‐67, and is concerned with maintenance of the high proliferative activity in cancer cells with aggressive proliferation.

Focal Atrophy of the Seminiferous Tubule in the Human Testis

Three types of focal atrophy of the seminiferous tubule, A, B and A+B type, were recognized in the 250 testes obtained from human subjects who died of acute death. The A‐type consists of complete hyalinized scarring of the tubule and the B‐type of varing degrees of tubular collapse. The A+B type shows combined atrophy of A and B‐type in which B‐type occurs peripherally to A‐type occlusion in sequence and might be induced by local obstruction of luminal flow. Focal tubulitis is uncommon but has been observed in the A+B type atrophy which is characterized by sperm retention and the presence of sperm‐phagocytic macrophages in the lumen. Areas around the rete testis and periseptal zones of the testis are preferential sites for tubular atrophy. B‐type atrophy as an abortive form of A‐type is predominant in the younger groups of the second to third decades, and the incidence of A‐type increases after the fifth decade. It is suggested that A and B‐type are brought about by locally limited ischemia in relation to circulatory disturbance with arteriolar hyalinosis which commonly appears focally in the postpuberal testis.