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It is challenging to demonstrate the capsule of Streptococcus pneumoniae (S. pneumoniae) under immunoelectron microscopy because of the thick mucopeptide cell wall hampering proper fixation. A novel rapid freeze fixation method was established to observe the capsule of S. pneumoniae. A strain of serotype 3 of S. pneumoniae isolate was analyzed after rapid freezing. An ethanol freezing-substitution fixing method was applied and immunohistochemical staining with osmium tetroxide was tested. The capsule was confirmed using the serotype 3 specific polyclonal antibodies labeled with colloidal gold particles. To the best of our knowledge, this is the first report of S. pneumoniae capsule by immunoelectron microscope.

Metastatic calcification due to hypercalcemia in adult Tcell leukemia-lymphoma (ATL) associated with osteolytic change for activation of osteoclasts are reported. These cases of serum calcium were at a high level, 15.1 and 19.4 mg/dl (normal range 8.4-10.4 mg/dl). Metastatic calcification was detected in the tubules of kidneys, in the pulmonary alveolar septa of lungs, in the myocardium, in the muscular layer of stomach, in the lower portion of media of aorta, in the mucosa of stomach, in the tubules of testis, and in the liver by von Kossa's silver nitrate method for calcium. Scattered osteoclasts were seen around the cortex of the bone. ATL cells stained with parathyroid hormone-related protein (PTHrP) by immunohistochemical procedure. Thus, PTHrP is an important bone resorption-stimulating factor of hypercalcemia in ATL. We have investigated the incidence of acute pancreatitis in ATL, and in the other diseases. Of the 317,325 autopsy cases, 632 were ATL; the numbers of the acute pancreatitis cases of them were 1,833 (0.58%) and 25 (4.0%), respectively. The odds ratio from 2 x 2 table is 7.17 and Peason's Chi-square statistics with one degree of freedom is 126, which is highly significant. Therefore, it was suggested that there is a close correlation between acute pancreatitis and hypercalcemia in patients with ATL. We proposed a new theory for the correlation between acute pancreatitis and hypercalcemia in ATL that all the hypercalcemic patiens exhibited high levels of nephrogenous cyclic adenosine monophosphate (NcAMP) stimulating pancreatic secretion in the extralobular ductal system of the pancreas and thus resuling in acute pancreatitis due to occlusion of the pancreatic duct., Acta medica Nagasakiensia. 1998, 43(3-4), p.26-33

Human T-cell leukemia virus type-1 (HTLV-1) is the causative agent of adult T-cell leukemia (ATL). Hypercalcemia is common in patients with ATL. These patients rarely develop metastatic calcification and acute pancreatitis. The underlying pathogenesis of this condition is osteoclast hyperactivity with associated overproduction of parathyroid hormone-related protein, which results in hypercalcemia in association with bone demineralization. The discovery of the osteoclast differentiation factor receptor activator of nuclear factor-κB ligand (RANKL), its receptor RANK, and its decoy receptor osteoprotegerin (OPG), enhanced our understanding of the mechanisms of ATL-associated hypercalcemia. Macrophage inflammatory protein-1-α, tumor necrosis factor-α, interleukin-1, and interleukin-6 are important molecules that enhance the migration and differentiation of osteoclasts and the associated enhanced production of RANKL for osteoblast formation. In this paper, we focus on metastatic calcification and acute pancreatitis in ATL, highlighting recent advances in the understanding of the molecular role of the RANKL/RANK/OPG system including its interaction with various cytokines and calciotropic hormones in the regulation of osteoclastogenesis for bone resorption in hypercalcemic ATL patients.

Immunohistochemical investigation was carried out to examine the characteristics of nasopharyngeal carcinoma (NPC) using 12 biopsy cases obtained from southern China. These cases were divided into three groups according to their predominant pattern associated with the cell and the tissue differentiation using the World Health Organization (WHO) classification as follows: (i) 4 cases with squamous cell carcinoma, (ii) 4 cases with differentiated nonkeratinizing carcinoma, (iii) 4 cases with undifferentiated carcinoma. These three types of NPC cases were reacted with keratin and epithelial membrane antigen (EMA). All neoplastic tissues were stained with keratin stronger than EMA. Squamous cell carcinoma and differentiated non-keratinizing carcinoma cases were diffusely stained strongly positive by immunoperoxidase for keratin. On the other hand, most of undifferentiated carcinoma cases were weakly or negatively reacted with keratin immunohistochemistry, and a part of tumor cytoplasms were stained strongly positive by keratin immunoreaction. For EMA immunoperoxidase method, squamous cell carcinoma cases were diffusely positive and most of differentiated non-keratinizing carcinoma and undifferentiated carcinoma cases were partly positive. However, none of NPC cases reacted with immunoreactivity for laminin 200, laminin 400, fibronectin, collagen, laminin receptor, and laminin/collagen receptor. Therefore, immunohistochemical characteristics of NPC showed epithelial cell origin, arising from squamous cells in nasopharynx., Acta medica Nagasakiensia. 1993, 38(2-4), p.182-185

It is difficult to propose simple idea in which cell adhesion molecules can he related to tumor growth and metastasis. The integrins function as extracellular matrix receptors binding to such proteins as collagen, laminin, and fibronectin. In this study, the cell culture of human glioma cells were xenografted to nude mice and rats associated with laminin injection, resulting in tumor growth and metastasis in the large amount of laminin group. In nude mouse experiment, cultured glioma cells were xenografted in the back of cutaneous with 10μg and 100μg laminin. The tumor lump of the 100μg laminin group grew faster and larger than the 10μg laminin group. In nude rat experiment, cultured glioma cells were xenografted intravascularly in the tail with l0μg and 100μg laminin. Of the 6 cases, 4 (66.7%) showed metastasis in the lungs after 2 weeks in the 100μg laminin group. However, metastasis was not detected in 10μg laminin group. These results suggest that quantities of laminin may play an important role in tumor growth and metastasis., Acta medica Nagasakiensia. 1993, 38(2-4), p.164-167

Goodpasture's syndrome is characterized by pulmonary associated with renal abnormalities and a group of systemic diseases share immunopathogenic mechanisms which is an autoimmune antiglomerular basement membrane disease. Pulmonary renal syndrome is rather common in the western countries, but rare in Japan. The present two cases showed clinical and pathological features of rapidly progressive glomerulonephritis complicated with massive pulmonary hemorrhage. Most of the hemorrhage appeared acute with intact red blood cells in alveolar spaces, and also had variable numbers of hemosiderin-laden macrophages as an evidence of old hemorrhage. In one case of Goodpasture's syndrome, linear deposits of IgG were demonstrated in alveolar walls, glomerular capillary, Bowman's capsular basement membrane, and tubules of kidneys. These evidences indicate the present two cases to be the so-called Goodpasture's syndrome., Acta medica Nagasakiensia. 1993, 38(2-4), p.49-53

Using cultured glioma cell line, we demonstrated in vitro and in vivo that when low grade glioma was pass through glioma cell culture at 19th passage, these tumor cells were malignantly transformed. This cell line was established from tissue culture of neurosurgical specimen, pathologically diagnosed as glioma (low grade glioma; grade 1-11) with glial fibrillary acidic protein (GFAP) positive but vimentin negative. When glioma cell cultured at 19th passage was injected into xenograft nude mice, histopathological and immunohistochemical analysis demonstrated that low grade glioma cells were transformed to high grade glioma (grade IV). High grade glioma cell can produce vimentin, laminin and fibronectin, but can not produce GFAP. It is suggested that vimentin and GFAP are useful and important marker in differentiating low grade glioma from high grade glioma. The mechanism of malignant transformation of low grade glioma cells is not clear. The possible explanation may be that the low grade glioma gene include malignant potential originally, as a result, low grade glioma cells change gradually to high grade glioma., Acta medica Nagasakiensia. 1993, 38(2-4), p.79-82

Histopathological analysis was carried out to examine the characteristics of nasopharyngeal carcinoma (NPC) using 38 biopsy cases obtained from southern China and 34 cases obtained from Japan. These cases were divided into three groups according to their predominant pattern associated with the cell and the tissue differentiation using the World Health Organization (WHO) histological classification of tumors as follows: (i) squamous cell carcinoma; 6 Chinese cases and 5 Japanese cases, (ii) differentiated nonkeratinizing carcinoma; 25 Chinese cases and 7 Japanese cases, (iii) undifferentiated carcinoma; 7 Chinese cases and 22 Japanese cases. The age of Chinese patients ranged from 21 to 80 years with a mean of 46 years and Japanese patients ranged from 17 to 89 years with a mean of 53.8 years. The male to female ratio was 2.7:1.0 in China and 1.8:1.0 in Japan. The most common histological type of NPC in China was differentiated non-keratinizing carcinoma, however, it was undifferentiated carcinoma in Japan. The mechanism of cancer arising from the nasopharynx may be quite different between Chinese and Japanese cases. The Chinese NPC cases were considered as continuity shift from normal epithelium to differentiated nonkeratinizing carcinoma, to undifferentiated carcinoma sequence. On the other hand, the Japanese cases were considered discontinuity shift. Therefore, in Chinese cases, NPC formation mechanism may be that EBV infection and chemical carcinogens can act synergistically in the enhancement of both viral replication and malignant transformation. It is suggested that some Japanese cases are associated with Epstein- Barr virus infection, although, mechanism of formation of NPC in Japanese cases is not yet clear., Acta medica Nagasakiensia. 1993, 38(2-4), p.242-246

Proliferative activity of nasopharyngeal carcinoma (NPC) using 20 biopsy cases obtained from southern China were analyzed by immunolabelling with the monoclonal antibody of proliferating cell nuclear antigen (PCNA), the numbers of interphase silver stained nucleolar organizer regions (AgNORs), and mitotic indices. Nucleolar organizer regions (NORs) are loops of ribosomal DNA (rDNA) occurring in the nucleoli of cells, which transcribe to ribosomal RNA (rRNA) and ultimately direct ribosome and protein formation. PCNA is a nuclear protein synthesized in the nucleus late in G1 phase, peaking during S phase and subsiding during G2 and M phases of the cell cycle. These may indicate that PCNA and AgNORs are present at different stages of the cell cycle. The authors found that 28% of differentiated non-keratinizing carcinoma type of NPC and 18% of undifferentiated carcinoma type of NPC were positively stained by PCNA. The mean AgNOR counts for differentiated non-keratinizing carcinoma type of NPC, and undifferentiated carcinoma type of NPC were 1.8 and 1.2, respectively. The mean mitotic counts for differentiated non-keratinizing carcinoma type of NPC and undifferentiated carcinoma type of NPC were 3.4 and 2.1, respectively. Therefore, it is suggested that PCNA labelling immunoreactivity may be correlated with AgNORs score and mitotic indeces, thus in NPC, these number and size have been regarded as markers of cell proliferative ability., Acta medica Nagasakiensia. 1993, 38(2-4), p.200-205

Immunohistochemical analysis was carried out to examine infiltration of lymphocytes in the tumor nests of nasopharyngeal carcinoma (NPC) using 38 biopsy cases obtained from southern China. These cases were divided into 3 groups according to their prominent pattern associated with the cell and the tissue differentiation using the World Health Organaization (WHO) classification as follows: (i) 6 cases of squamous cell carcinoma (16%), (ii) 25 cases of differentiated non-keratinizing carcinoma (66%), (ii) 7 cases of undifferentiated carcinoma (18%). All tumor tissues reacted diffusely with MB-1, however, these did not react with L26 (CD20), 4KB5 (CD45R), MT-1, and leukocyte common antigen (LCA). In the tumor nests, normal lymphocytes were observed in squamous cell carcinoma type (50%), differentiated non-keratinizing carcinoma type (72%), and undifferentiated carcinoma type (86%). Therefore, it is suggested that a large number of infiltrating non-neoplastic T-cells may be related to good prognosis. T-cell-rich B-cell lymphomas (TCRBCLs) are recently described as unusual non-Hodgkin's lymphomas associated with predominance of reactive T-cells. Therefore, the mechanism of T-cell lymphocyte infiltration adjacent to the tumor cell is considered that these T-cell infiltrates may be the passive response to cytokine secretion by the tumor cells., Acta medica Nagasakiensia. 1993, 38(2-4), p.189-192

The risk of occupational transmission of human immunodeficiency virus (HIV) to health care workers with intensive exposure to HIV infected patients was considered, because the author autopsied an acquired immunodeficiency syndrome (AIDS) patient in February 1990. A few case reports suggested that the risk of HIV infection incurred when health care workers are exposed to blood or body fluids of HIV infected patients through accidental needle-stick exposure or contamination of an open wound or mucous membrane. For the prevention of AIDS, the pathologists and staff must use double-gloves, gown, mask, apron, shoes cover, medical cap, undershirt, pant, and goggle when in contact with AIDS patient-related materials during postmortem procedure and treatment of surgical speciments., Acta medica Nagasakiensia. 1991, 36(1-4), p.85-88

Two cases of metastatic calcification due to hypercalcemia in adult T-cell leukemia-lymphoma (ATLL)associated with osteolytic change for activation of osteoclasts are reported. These cases of serum calcium were at a high level, 16.2 and 19.4mg/dl (normal range 8.4-10.4mg/dl). In our cases, metastatic calcification was detected in the tubules of kidneys, in the pulmonary alveolar septa of lungs, in the myocardium, in the muscular layer of stomach, in the lower portion of media of aorta, in the mucosa of stomach, in the tubules of testis, and in the liver by von Kossa's silver nitrate method for calcium. Scattered osteoclasts were seen around the cortex of the bone. Roentgenograms showed osteolytic change in the skull, in the bilateral ulna, in the radius, in the humerus, in the tibia, and in the fibula. Therefore, hypercalcemia in ATLL may be caused by bone resorption-stimulating factors which promote the differeniation of osteocalast cells, resulting in calcium increases in the serum., Acta medica Nagasakiensia. 1991, 36(1-4), p.24-28