新里 昌功

After finding tonsil-like structures near the entrance of vagina of a laboratory shrew (Suncus murinus), which we subsequently designated as vaginal tonsils, we performed detailed immunohistochemical and developmental studies. The location of T and B cells in the vaginal tonsils differed from that in the palatine tonsils or that in the lymphoid nodes of other animals. The boundary between the germinal center region and the region encompassing follicular interfollicular tissue was not clear. B cells were widely distributed and very dense in the parenchyma, but they were scattered in the epithelial area (B cells were present in around 90% of the vaginal tonsil tissue). In contrast, T cells were scattered in the parenchyma and in the epithelial area (T cells were present in around 10% of the vaginal tonsil tissue). B cells were more prominent than T cells throughout the development of these structures and the epithelium was invaded by many immigrating cells. The size of the vaginal tonsils changed during postnatal development. Vaginal tonsils are structurally similar to other tonsils, and they may function to protect the vagina from infection.

High temperature- and pressure-treated garlic (HTPG) has been shown to have enhanced antioxidative activity and polyphenol contents. Previously, we reported that HTPG inhibited 1,2-dimethylhydrazine-induced mucin depleted foci (premalignant lesions) and O-6-methylguanine DNA adduct formation in the rat colorectum. In the present study, we investigated the modifying effects of HTPG on N-ethyl-N'-nitro-N-nitrosoguanidine (ENNG)-induced pyloric stomach and small intestinal carcinogenesis in mice. Male C57BL/6 mice were given ENNG (100 mg/l) in drinking water for the first 4 weeks, then a basal diet or diet containing 2% or 5% HTPG for 30 weeks. The incidence and multiplicity of pyloric stomach and small intestinal (duodenal and jejunal) tumors in the 2% HTPG group (but not in the 5% HTPG group) were significantly lower than those in the control group. Cell proliferation of normal-appearing duodenal mucosa was assessed by MIB-5 immunohistochemistry and shown to be significantly lower with 2% HTPG (but again not 5% HTPG) than in controls. These results in dicate that HTPG, at 2% in the diet, inhibited ENNG-induced pyloric stomach and small intestinal (especially duodenal) tumorigenesis in mice, associated with suppression of cell proliferation.

We investigated the distribution of T lymphocytes, B lymphocytes, and S-100 protein-immunoreactive dendritic-like in the anal tonsil of the laboratory shrew, Suncus murinus. In adult animals, T lymphocytes were located mainly at the periphery of the anal tonsil, especially around small blood vessels. B lymphocytes were located in the central and subepithelial region of the anal tonsil, which includes primary lymphoid follicles, and in which there are small numbers of scattered T lymphocytes. B and T lymphocytes were distributed over 72.7 and 27.3% of the tonsillar area, respectively. However, their areas of distribution were not clearly distinguished. The areas containing B lymphocytes were enriched in S-100 protein antibody-immunoreactive cells, which exhibited a dendritic shape. These S-100-positive cells appeared to be identical to the follicular dendritic cells (FDC) seen in the follicles of lymphoid organs. These results suggest that the anal tonsils constitute one of the gut-associated lymphoid tissues (GALT), and that a function of the anal tonsil includes the capture of intruding antigens that would generate protective antibody responses. Microsc. Res. Tech. 74: 819-824, 2011. (C) 2010 Wiley-Liss, Inc.

We studied the role of osteopontin (OPN) in host responses against rotavirus (RV) infection. OPN knockout (OPN-KO) suckling mice were more susceptible to RV (strain EW) infection and showed prolonged diarrhea duration compared to wild-type (WT) suckling mice. OPN in the small intestine of WT mice was expressed after 48 h post-infection. On day 2 postinfection, mRNA levels of interleukin-1 beta, tumor necrosis factor-alpha, and interleukin-15 in OPN-KO mice were lower than in WT mice, although mRNA expression of Th-1- and Th-2-related cytokines in the small intestine were nearly the same between OPN-KO and WT mice. These results suggested that OPN is involved in innate responses against RV infection.

It is well known that avian yolk sac is involved in both primitive and definitive erythropoiesis during embryonic development. Definitive erythropoiesis is first detected at about 4-5 days incubation and its maximum activity is reached between day 10 and 15 of incubation, ending between days 18 and 20 of incubation. We confirmed the definitive erythropoietic foci in the chicken yolk sac throughout the 5th to 19th day of incubation by histochemical light and electron microscopy. The definitive erythropoietic foci were observed in the yolk sac endodermal layer from day 5 until day 19, just before hatching. Ultrastructurally, definitive erythropoietic foci were observed extravascularly in the yolk sac endodermal cell layer in direct contact with the vitellolysis zone. These findings provide a basis for clarifying definitive erythropoiesis in vertebrates.

Light and electron microscopic examination of first-trimester and term human placental tissues were performed to identify erythrocytes containing hemoglobin in the villous trophoblast cell layer. Erythrocytes were not identified in chorionic villous epithelium at week 7 of gestation. These cells first appeared in the villous cytotrophoblast at week 8, and continued to be present in the villous cytotrophoblast until week 9, as shown by benzidine staining. At week 12 gestation, a cluster of erythrocytes was present in a villous syncytial sprout. At 40 and 41 weeks gestation, erythrocytes were located in the villous cytotrophoblast cell layer. Electron microscopic observations focused on the cytoplasm of villous cytotrophoblast at week 8, the syncytial sprout at week 12 and the cytotrophoblast cell layer at term, confirmed the presence of erythrocytes at an extravascular location, as observed by light microscopy.

Osteopontin (OPN) knockout mice (OPN-KO mice) died of Plasinodium chabaudi chabaudi infection, although wild-type (WT) mice had self-limiting infections. OPN was detected in the WT mice at 2 days postinfection. OPN-KO mice produced significantly smaller amounts of interleukin-12 and gamma interferon than WT mice produced. These results suggested that OPN is involved in Th1-mediated immunity against malaria infection.

We examined the modifying effect of freeze-dried whole-leaf Aloe arborescens Miller var. natalensis Berger (Kidachi aloe in Japan designated as 'ALOE') on azoxymethane (AOM)-induced intestinal carcinogenesis in rats. Male F344 rats (4 weeks old) were fed basal diet or experimental diet containing 0.2% or 1% ALOE for 28 weeks. Starting two weeks later, the animals received subcutaneous injections of AOM once weekly for 10 weeks. The incidence of colorectal adenocarcinomas in the 0.2% (but not 1%) ALOE group showed a strong tendency for decrease (p = 0.056) from the control group. Further, the adenocarcinoma incidence in the entire intestine (small and large intestines) in the 0.2% ALOE group was significantly (p = 0.024) decreased compared to the control value. However, there were no significant differences in tumor multiplicities of colorectal or entire intestines among the 3 groups. In addition, we also studied the safety of long-term ingestion of ALOE as a health food or natural thickening stabilizer. Rats were fed the basal diet or 1% ALOE diet for 35 weeks without AOM treatment. Feeding with 1% ALOE did not affect most hematological and serum biochemical parameters in the rats. These results indicate that a low level of ALOE ingestion might have a mild suppressive effect on intestinal tumor growth without harmful side effects.

Common subungual tumors include fibrokeraroma, Koene's tumor, glomus tumor, epidermoid cyst, Bowen's disease and squamous cell carcinoma.(1) Solitary neurofibromas in this area are rare. Subungual neurofibromas normally grow without obvious symptoms. As nails conceal subungual tumors, only a proportion of these tumors are visible. We suggest that subungual neurofibromas might have been underreported. A rare case of solitary subungual neurofibroma not associated with von Recklinghausen's disease is reported. Only a few cases of solitary neurofibroma have previously been reported in the literature. Other cases of this tumor might go unnoticed due to lack of symptoms.

We previously observed Langerhans cells (LC) in the squamous metaplastic urinary bladder in rats fed a vitamin A-deficient diet. Here we report the finding of LC in the iliac lymph nodes, one of the abdominal lymph nodes draining to the urinary bladder. LC had previously only been observed in the superficial and hilar lymph nodes, but never in the abdominal lymph nodes. This is also the first observation of Birbeck granule (BG)-positive LC in the iliac lymph nodes. LC were not found in the transitional mucosa of the urinary bladder. In the present experiments, the LC were observed in both the squamous metaplastic mucosa and lamina propria of the urinary bladder, and also in the iliac lymph nodes, which drain into the urinary bladder. BG-positive LC may mature not only in the squamous epithelia, but also in squamous metaplastic mucosa. LC in the lamina propria of the urinary bladder might migrate to the iliac lymph nodes, which are regional lymph nodes, where they could function in antigen presentation. LC in the skin play a role in antigen-antibody interactions, and the squamous metaplastic mucosa of the urinary bladder may be a similar environment, in which LC in the mucosa can migrate to the regional lymph nodes.

We examined the modifying effects of freeze-dried whole-leaf Aloe arborescens Miller var. natalensis Berger (Japanese name, Kidachi aloe; designated as 'ALOE') on 1,2-dimethylhydrazine (DMH)-induced colorectal tumorigenesis in mice. Female ICR mice (7-weeks old) were given a basal diet or a diet containing 1, 0.5 or 0.1% ALOE for 32 weeks. One week later, all mice were injected i.p. with DMH (20 mg/kg, once weekly for 10 weeks) or vehicle (1 mM EDTA solution, pH 6.5). At 32 weeks, animals were killed by exsanguination, and the colorectums were processed for histological examination. The administration of ALOE (1, 0.5 or 0.1% in diet) did not induce diarrhea or reduction of body weight. In mice given DMH and 1% ALOE (Group 2), the incidence and multiplicity of colorectal adenomatous hyperplasias were significantly decreased as compared with mice given DMH alone (Group 1) (both p < 0.05), whereas the incidence and multiplicity of tumors (adenoma and adenocarcinoma) in Group 2 tended to be lower than those in Group 1. In addition, the incidence and multiplicity of the colorectal proliferative lesions (the total of adenomatous hyperplasias, adenomas and adenocarcinomas in mouse colorectum) in Group 2 were significantly decreased as compared with Group 1 (both p < 0.01). No colorectal proliferative lesions were found in animals that did not receive DMH. These results indicated that ALOE reduces the incidence and multiplicity of DMH-induced colorectal proliferative lesions, especially adenomatous hyperplasia, in mice.

Approximately 30 cases of carcinoid tumor of the kidney have been reported in the English literature, including three cases found as components of teratomas. Renal composite tumors associated with somatostatinoma have not been described. A 53-year-old female presented with an incidentally found right renal cystic lesion. Computed tomography demonstrated a cystic lesion associated with a solid nodule in the right kidney and postcontrast dynamic MRI revealed enhancement of the solid nodule. The patient underwent radical nephrectomy for the kidney lesion and is now well without recurrence 21 months after the operation. From the histopathological findings we diagnosed the cystic lesion as a composite tumor composed of mucinous cystadenoma and carcinoid tumor. Immunohistochemistry demonstrated the majority of cells of in carcinoid portion to be positive for antisomatostatin staining. The present case is the first documented composite tumor of mucinous cystadenoma and somatostatinoma of the kidney.

The mouse visceral yolk sac (VYS) is widely known to play an important role as erythropoietic tissue during embryonic periods. Mouse VYS from embryonic days 9 to 12 was examined by light microscopy, electron microscopy and histochemical analysis with benzidine to detect the presence of hemoglobin with special reference to the development of VYS, the disappearance of the blood islands in VYS, and the appearance of a novel structure in the form of erythrocyte-like globules in VYS endodermal cells. The villous appearance of VYS became complicated by the development of VYS endodermal cells. The blood islands positive for the benzidine reaction were light microscopically detected on embryonic days 9, 10, and 11. They disappeared on embryonic day 12, however. Erythrocyte-like globules positive for the benzidine reaction were not observed in VYS endodermal cells on embryonic days 9, 10, and It, but then appeared on embryonic day 12, by light and electron microscopy. Erythrocyte-like globules in VYS endodermal cells, which appear after the disappearance of blood islands in VYS, may participate in erythropoiesis during embryonic development.

Background: Generally, Langerhans cells deliver antigen information from the skin to the draining lymph nodes via lymph vessels. Methods: By immunohistopathology, we investigated the delivery route of Langerhans cells in human skin using CD1a and S-100 protein antibodies. Results: We noted CD1a- and S-100-positive Langerhans cells in the lymph vessels of the dermis. These were shaped like dendritic cells and presented with some lymphocytes, melanophages, melanin granules and lymph in the same vessels. Conclusion: These observations support the concept that Langerhans cells deliver antigen peptides to regional lymph nodes via afferent lymph vessels. Copyright (C) 2003 S. Karger AG, Basel.

Hematological studies have revealed the importance of the visceral yolk sac (VYS) in the primitive erythropoiesis of mouse embryos at an early stage before day 12. We examined the possibility of the occurrence of extra-embryonic erythropoiesis at a stage later than embryonic day 12 by light and electron microscopic analyses. Surprisingly, a novel structure in the form of erythrocyte-like globules was observed in the VYS endodermal cells. They were consistently present in the VYS endodermal cells from embryonic day 12 until day 18 (birth is day 19), by immunocytochemical and enzyme histochemical analyses. They were immuno-positive for mouse erythrocyte antibody and also positive for the benzidine reaction showing the presence of hemoglobin. The erythrocyte-like globules were shown to be the erythrocytes present in the cytoplasm. These results indicated that erythropoiesis in the VYS endodermal cells continues from the early embryonic stage, as primitive erythropoiesis, until the late stage.

A 73-year-old man who had been undergoing chronic haemodialysis (CHD) for 3 years developed haemophagocytic syndrome (HPS) that might have been triggered by Epstein-Barr virus (EBV) infection. The patient finally died of miliary tuberculosis (TB) reactivation that promoted the progression of HPS. Immunological abnormalities in patients undergoing CHD may be notable. The early diagnosis of TB reactivation may be important for reducing the mortality in cases of HPS, as a high incidence of TB is encountered in patients undergoing CHD. In contrast, the simultaneous occurrence of an EBER-positive hybridization signal with T cell-specific immunolabelling of CD45RO cells was well detected in the spleen and lymph nodes, and interferon gamma was elevated in the serum. These findings coincide with the reported preferential expansion of T cells rather than B cells in EBV infection, and support the hypothesis that systemic hypercytokinaemia caused by the proliferation of EBV-infected T cells may play a crucial role in the development of HPS.

The dermal histology and the regional draining superficial lymph node of a new mutant strain of hairless rats (ISh) were investigated. The homozygote ISh rat was characterized as having naked and wrinkled skin. The comedo-like cysts in the skin resembled human acne and vulgaris. Histopathologically, many Langerhans' cells positive for anti-protein kinase C type II antibody (PKC-II) were recognized in the epidermis. The superficial lymph nodes were significantly larger than those of the heterozygote. The paracortex of these lymph nodes was expanded and the number of Langerhans' cells mainly increased in these areas. These lymph node lesions were similar to those of human dermatopathic lymphadenopathy. The ISh rats should be a very useful animal model for studying dermatopathic lymphadenopathy in humans. Furthermore, they may be a valuable animal model for investigating the mechanisms of not only maturation, movement and migration of Langerhans' cells, but also of their function for antigen presentation.

In the present study, immunohistochemical and immunoelectron microscopic techniques were used to differentiate Langerhans cells (LC) from interdigitating cells (IDC) in the lymph nodes (LN) of dermatopathic lymphadenopathy. The majority of the dendritic cells that existed in the LN of dermatopathic lymphadenopathy were positive for OKT-6 (CD 1a) antibody. It was concluded that these dendritic cells were not IDC, but LC. Electron microscopically, LC in these LN contained a few Birbeck granules (BG). In order to prove the fact that these dendritic cells were LC, the existence of BG was investigated ultrastructurally by examining serial sections, and immunoelectron microscopically for CD 1a positive cells. Most of the LC in the lymph nodes we examined were negative for the anti-proliferating nuclear antigen (PCNA) antibody. This finding may mean that LC in the LN are fully developed cells and do not divide in the LN. Langerhans cells may migrate from the skin lesions to the paracortical areas in the LN, which then may become enlarged.

Langerhans cells (LCs) in the squamous metaplastic epithelium of the trachea and the urinary bladder of vitamin A-deficient rats were examined. Both round, immature cells which were considered to be precursor LCs, and mature LCs with prominent dendritic processes, were recognized in the present study. Both cells were positive for anti-protein kinase type II antibody (PKC-II), and although the former did not have Birbeck granules (BGs), the latter did. The precursor LCs without BGs first appeared in the early stage of squamous metaplasia. As the differentiation of squamous metaplasia progressed, the number of BG bearing LCs increased, and the precursor cells became mature LCs in the later stage of squamous metaplasia of both the trachea and the urinary bladder. In the later stage of squamous metaplasia of the urinary bladder, LCs with BGs also appeared in the lamina propria. These findings indicate that precursor cells which probably migrate from the bone marrow, disperse into the metaplastic squamous epithelium in morphologically immature forms, and mature into BG bearing LCs as a result of contact with squamous epithelium.

OBJECTIVE: To successively examine intranuclear inclusions and nuclear grooves in the same papillary thyroid cancer specimens using a light microscope (LM), scanning electron microscope (SEM) and transmission electron microscope (TEM). STUDY DESIGN: We stained cells by the Papanicolaou method after fixation in 1.25% glutaraldehyde for LM and then attempted to observe them successively by SEM-TEM after fixation in 2% paraformaldehyde and 2% osmium tetroxide. RESULTS: On SEM, intranuclear inclusions were observed as elevated parts, like hills, and nuclear grooves were observed as deep fissures or shallow cracks, sometimes with a few in one cell. On TEM, both intranuclear inclusions and nuclear grooves seemed formed by the nuclear membranes. Intranuclear inclusions also possessed cytoplasm and/or cytoplasmic organelles within some expanded areas in the nuclear grooves. CONCLUSION: In was evident from our three-step technique that intranuclear inclusions and nuclear grooves were essentially the same structures. © 1996 The International Academy of Cytology Acta Cytologica.

The present study shows that Langerhans cells can be differentiated from interdigitating cells at the light microscopic level. Superficial lymph nodes and skin taken from necropsies and the lymph nodes of dermatopathic lymphadenopathy (DPL) were used for this experiment. Sections of lymph node and skin were embedded using the acetone, methyl benzoate and xylene (AMeX) method and dendritic cells were immunostained with anti S-100 protein antibody (S-100, and OKT-6 (CD1a) using the restaining method. Langerhans cells in the skin were positive for both CD1a and S-100. Dendritic cells positive for both CD1a and S-100, and dendritic cells positive for S-100, but not for CD1a were observed in superficial lymph nodes. In normal superficial lymph nodes, there were more interdigitating cells than Langerhans cells. The majority of the dendritic cells in the DPL were Langerhans cells. We conclude that the S-100 and CD1a positive cells are Langerhans cells, and the S-100 positive-CD1a negative cells are interdigitating cells.

The identification of cells in body cavities of cancer patients is sometimes difficult to make. In order to make a definite cytological diagnosis, we observed the same cells by using light microscopy (LM)-scanning electron microscopy (SEM)-transmission electron microscopy (TEM). In this study we first stained cells by the Papanicolaou method after fixation in 1% glutaraldehyde for LM, and then attempted to observe them successively by SEM-TEM after fixation in 1% paraformaldehyde and 1.25% OsO4. Our method and procedures in examining successively one and the same cells in body cavity fluids by using LM, SEM, and TEM ensured accurate discrimination among adenocarcinoma cells, mesothelial cells, and macrophages. The results of this study suggest that LM-SEM-TEM may be of diagnostic value in distinguishing among mesothelial cells, macrophages, and adenocarcinoma cells. This method also succeeded in disclosing differences between the ultrastructure of the cell surfaces, and those of the cytoplasm, and of the nuclei. It is desirable that LM-SEM-TEM observation can be introduced into various aspects in order to obtain an improvement in the diagnosis by cytologic examination, the judgment of therapeutical effects, drug selection, and prognostic presumption. (C) 1994 Wiley-Liss, Inc.

The migration and maturation of Langerhans cells (LCs) in rat tracheal squamous metaplasia due to vitamin A deficiency were investigated immunohistochemically and electron microscopically. In the early stage of metaplasia, i.e. basal cell hyperplasia, no LCs with Birbeck granules (BGs) could be found, but there were desmosome-free cells which had the morphological charcteristics of immature LCs. They were clearly different from inflammatory cells such as macrophages and lymphocytes, and were, therefore, considered to be precursors of LCs. In the stage of stratification, small numbers of Ia- and protein kinase C type II (PKCII)-positive cells were recognized. Ultrastructually they were immature LCs with ovoid nuclei, many free ribosomes and few dendrites. The cytoplasm was dark and a few BGs and atypical granules (AGs) could be seen in the Golgi area. In the early stage of cornification, LCs with partially intended nuclei, prominent nucleoli and well-developed Golgi complexes were found. There were many BGs and AGs and structures transitional between them in the Golgi areas. In epithelium showing mature squamous metaplasia, many Ia- and PKCII-positive dendritic cells could be seen. Most of these were typical mature LCs with lobulated nuclei, clear cytoplasm and prominent dendritic processes. The number of BGs and AGs was fewer than in the LCs found in the early stage of cornification, and these granules were distributed throughout the cytoplasm. In the final stage, where the basal cells had differentiated into a flatter epithelium, few LCs could be seen. These findings suggest that the precursors of LCs without BGs migrate into metaplastic squamous epithelium and mature into LCs forming BGs after exposure to the microenvironment of the squamous epithelium.