Expression of estrogen receptor alpha (ER alpha) in the rat temporomandibular joint

Numerous epidemiological studies have pointed out a higher frequency of temporomandibular disorder (TMD) in women than in men, which indicates the involvement of a sex hormone, such as estrogen, in the pathogenesis of TMD. Although estrogen is known to play pivotal roles in osteoarthrosis or rheumatoid arthritis in systemic joints, there have been few reports about the role of estrogen in the temporomandibular joint (TMJ). The effect of estrogen is generally mediated by the estrogen receptors (ERs) ER alpha (the predominant type) and ER beta. In this study we examined the expression of ER alpha protein and mRNA in the TMJ of adult male rats by immunocytochemistry and in situ hybridization histochemistry. Intense ER alpha immunoreactivity was localized in the synovial lining cells, stromal cells in the articular disc, and chondrocytes in the TMJ. These ER alpha-immunopositive synovial lining cells are characteristic of cytoplasmic processes identified with confocal and immunoelectron microscopy, which indicates that they are synovial type B cells. In situ hybridization histochemistry confirmed intense signals for ER alpha in the synovial lining cells and the sublining fibroblasts at mRNA levels. The nuclei of chondrocytes showed an intense immunoreaction for ER alpha in the maturative and hypertrophic layers of the articular cartilage. In addition to the nuclear localization of ER alpha, a weak immunoreaction appeared in the cytoplasm of some ER alpha-positive cells. These findings support the hypothesis that TMJ tissue-at least in the male rat-has the potential to be an estrogen target tissue.     

Formation and distribution of the sural nerve based on nerve fascicle and nerve fiber analyses

The formation and distribution of the sural nerve are presented on the basis of an investigation of 31 legs of Japanese cadavers using nerve fascicle and fiber analyses. Nerve fibers constituting the medial sural cutaneous nerve were designated as 'T', whereas those constituting the peroneal communicating branch were designated as 'F'. In 74.2% of cases (23/31), the T and F fibers joined each other in the leg, whereas in 9.7% of cases (3/31) they descended separately. In 16.1% of cases (5/31), the sural nerve was formed of only the T fibers. The sural nerve gave off lateral calcaneal branches and medial and lateral branches at the ankle. The lateral calcaneal branches always contained T fibers. The medial branches consisted of only T fibers, whereas most of the lateral branches consisted of only F fibers (71.0%; 22/31). In addition to the T and F fibers, P fibers, which derived from the superficial and deep peroneal nerves, formed the dorsal digital nerves. The P fibers were entirely supplied to the medial four and one-half toes. However, they were gradually replaced by the T and F fibers in the lateral direction. The 10th proper dorsal digital nerve consisted of T fibers only (38.7%; 12/31), of F fibers only (19.4%; 6/31) or of both T and F fibers (38.7%; 12/31). These findings suggest that the T fibers are essential nerve components for the skin and deep structures of the ankle and heel rather than the skin of the lateral side of the fifth toe. The designation of the medial sural cutaneous nerve should be avoided and only the T fibers are appropriate components for naming as the sural nerve.     

Reactive oxygen species-producing site in hydrogen peroxide-induced apoptosis of human peripheral T cells: involvement of lysosomal membrane destabilization

In our previous study, we examined the effect of exogenous hydrogen peroxide, which causes a potent oxidative stress and has been demonstrated to be a potent apoptosis-inducer in many kinds of cells. We found that the addition of 1 or 10 mM hydrogen peroxide induced reactive oxygen species (ROS) formation, oxidative DNA damage, dysfunction of the mitochondrial membrane potential, and early apoptotic changes in the human osteosarcoma cell line HS-Os-1. We therefore concluded that intracellular ROS formation was involved in the hydrogen peroxide-induced apoptosis of HS-Os-1 cells. In contrast to the osteosarcoma cell line HS-Os-1, human peripheral T cells are considered to be easily susceptible to oxidative stress, because these cells lack peroxidase activity. Therefore, in this study, we investigated the site of ROS formation by utilizing MitoCapture, H2DCFDA (succinimidyl ester of dichloro-dihydrofluorescein diacetate), DAPI (4',6-diamidino-2-phenylindole), and LysoSensor. Our results showed that ROS formation was apparently diffusely distributed in T cells oxidatively stressed with 0.1 mM hydrogen peroxide. Moreover, lysosomal swelling and deformity, possibly revealing lysosomal membrane destabilization, were observed in these cells. Based on the above results, there exists an apoptotic cascade involving early lysosomal membrane destabilization in the hydrogen peroxide-induced apoptosis of human peripheral T cells. Therefore, the possible involvement of lysosomal protease leakage caused by hydroxyl radical formation in lysosomes (possibly resulting in mitochondrial membrane dysfunction) is considered to play an important role in hydrogen peroxide-induced T cell apoptosis.     

Interferon-{beta} inhibits bleomycin-induced lung fibrosis by decreasing transforming growth factor-{beta} and thrombospondin

Pulmonary fibrosis is the result of abnormal processes of repair that occur after lung injury. Transforming growth factor (TGF)-beta is a key molecule in the progression of pulmonary fibrosis. Although clinical use of interferon (IFN)-beta did not improve survival in patients with idiopathic pulmonary fibrosis, because some preclinical studies have suggested that IFN-beta is a potent inhibitor of fibrogenesis, beneficial effects of IFN-beta have been expected. We therefore attempted to determine effects of IFN-beta and investigated the mechanism of action of IFN-beta in bleomycin-induced pulmonary fibrosis. Bleomycin at Day 0 and IFN-beta for 4 wk were administered intravenously to ICR mice. At 28 d after bleomycin injection, histologic and chemical analysis was performed for evaluation of effects of IFN-beta. Tissue distribution and amounts of TGF-beta1 and thrombospondin (TSP)-1/2 were analyzed. IFN-beta attenuated prolylhydroxylase activity, resulting in inhibition of pulmonary fibrosis. Bleomycin-induced increase in TGF-beta1 in epithelial cells and extracellular matrix was attenuated by IFN-beta. TSP-1/2 was limited in platelets of control mice, but was present in foamy cells in fibrotic regions induced by bleomycin. These findings suggest that the antifibrotic effect of IFN-beta is inhibition of TGF-beta and its activation via decrease in TSP-1/2 in lung tissue and change in location of TSP-1/2 from platelets to foamy cells.     

Immunohistochemical study of tyrosine-hydroxylase-positive cells and fibers in the chicken spinal cord.

Tyrosine hydroxylase (TH)-positive cells and fibers were examined by immunohistochemistry in the chick spinal cord. TH-positive cells, which were located in laminae I, V and X, were most frequently found in the rostral part of the cervical spinal cord, with fewer cells being found in more caudal levels of the spinal cord. TH-positive cells located in lamina X, which were bipolar in shape, were mainly found in regions lateral as well as just ventral to the central canal. They had processes reaching to the central canal. The terminals of these cerebrospinal-fluid-contacting cells were oval in shape, and were most frequently found at the ventral wall of the central canal. There were dense clusters of TH-positive fibers in lamina X. A meshwork-like structure of TH-positive fibers was found over the lateral wall of the central canal. A high density of TH-positive fibers was also found in the medial part of laminae V-VII. In lamina IX, small numbers of TH-positive fibers were observed in the lateral motor column of the brachial spinal cord, and in the medial and lateral motor columns of the lumbosacral spinal cord. However, within the medial motor column of the brachial spinal cord TH-positive fibers were densely distributed around somal as well as dendritic profiles. Similar to our previous observations on serotoninergic fibers. TH-positive fibers were also differentially distributed in the ventral horn of the chicken spinal cord: a high density of TH-positive fibers was localized to specific regions of the spinal motor nucleus.     

Alpha 1-antichymotrypsin is present in diffuse senile plaques. A comparative study of beta-protein and alpha 1-antichymotrypsin immunostaining in the Alzheimer brain.

Immunocytochemical examinations of the brains of patients with senile dementia of the Alzheimer type, Down''s syndrome, and Gerstmann-Sträusslar-Scheinker disease were performed to clarify the relationship between alpha 1-antichymotrypsin and senile plaque amyloids. Alpha 1-antichymotrypsinlike immunoreactivity was enhanced by protease digestion but not by formic acid pretreatment. Almost all of the diffuse plaques and some small amyloid deposits, which were widely distributed in the cerebral cortex and/or subcortical regions of SDAT brains, were labeled by alpha 1-antichymotrypsin immunostaining. All types of senile plaques, eosinophilic tangles, and some neurons and astrocytes were labeled by alpha 1-antichymotrypsin staining. Diffuse plaques in a Down''s syndrome frontal lobe and in a senile dementia of the Alzheimer type cerebellum also were labeled by alpha 1-antichymotrypsin immunostaining. However neither subpial amyloid deposits nor subcortical small amyloid deposits could be detected by alpha 1-antichymotrypsin immunostaining. Kuru plaques in a Gerstmann-Sträusslar-Scheinker disease cerebellum also were not labeled by alpha 1-antichymotrypsin immunostaining. These results suggest that alpha 1-antichymotrypsin is associated with the early to late stages of amyloid deposition and senile plaque formation in senile demential of the Alzheimer type.     

Experimental haemarthrosis in rhesus monkeys: morphometric, biochemical and metabolic analyses

The effects of a single episode of massive haemarthrosis in rhesus monkeys were studied. Autologous whole blood was injected into a femorotibial joint of 16 anaesthetized monkeys, equally divided into four groups and killed 7 days, 2, 3 and 6 months post-injection (PI). Synovial membrane and femoral articular cartilage were analysed morphometrically and articular cartilage was further analysed biochemically and metabolically. At 7 days PI, morphometric evaluation revealed a significant increase (P less than 0.05) in synovial membrane cellularity and synovial intimal thickness of injected joints versus control joints. This change was no longer evident 2 months PI. There was also an overall (n = 16) significant increase (P less than 0.05) in femoral articular cartilage cellularity in injected joints. The average chondrocyte lacuna area of injected joints was not statistically different from the control joints. Biochemical analyses of femoral articular cartilage revealed a significant decrease in hexosamine concentration (P less than 0.05) of injected joints. There was no significant difference between the injected and control joints in hydroxyproline or total protein concentration. Metabolic analyses revealed a significant increase (P less than 0.05) in cartilage collagenous protein production by injected joints compared with control joints. There were no significant differences in cartilage or secreted total protein production between injected and control joints. There were also no significant differences in cartilage or secreted proteoglycan production between joints. Morphometric evaluation of articular tissues following massive haemarthrosis has quantified a temporary hyperplastic reaction. A significant decrease in cartilage hexosamine concentration in haemarthrotic joints suggests this is a crucial biochemical event in the pathogenesis of blood-induced cartilage destruction.     

HIV infection of CD4-CD8+ T-cell line derived from patients with HAM

This studies have attempted human immunodeficiency virus (HIV) infection in four CD4+ or CD8+ T-cell lines derived from HTLV-I associated myelopathy virus (HAM) patients. Not only CD4+ cell line but also CD8+ cell line could be infected with HIV and CD4+ cell line showed a higher susceptibility than CD8+ cell line on HIV infection. HIV antigen in early stage after HIV inoculation was detected by using enzyme-linked immunosorbent assay (ELISA) rather than indirect immunofluorescence assay (IFA). HTLV-I producing CD4+ and CD8+ cell lines became to express two viral antigens (HTLV-I and HIV) after HIV inoculation. The results indicated that CD4-CD8+ T-cell line from patient with HAM can be infected with HIV. So that, we have found that other epitopes except for CD4 antigen may be associated with HIV infection.     

Effect of SA-103 on experimental allergic models in vivo and in vitro--comparison with disodium cromoglycate.

The anti-allergic action of N-[4-(4-methoxyphenyl)-2-thiazolyl]-1H-tetrazol-5-carboxamide (SA-103) was investigated and compared with that of disodium cromoglycate (DSCG). 1) Oral administration of SA-103 (0.1-10 mg/kg, p.o.) showed dose-dependent inhibition of 48 hr homologous passive cutaneous anaphylaxis (PCA) in rats. The inhibition rate (50%) of the compound at 1 mg/kg was comparable to that of DSCG at 1 mg/kg (i.v.). 2) Both of the drugs concentration-dependently inhibited the release of in vitro anaphylactic histamine from rat peritoneal exudate cells, but SA-103 was 1,000 times as potent as DSCG. 3) High doses (50 and 100 mg/kg, p.o.) of SA-103 tended to suppress 7-day homologous PCA in guinea pigs by only 20-30%. DSCG (100 mg/kg, i.v.) did not influence the reaction. 4) Neither anaphylactic histamine nor leukotriene release from guinea pig lung fragments was markedly influenced by SA-103 (10(-8)-10(-5) g/ml) or DSCG (10(-5)-10(-3) g/ml). 5) The histamine and serotonin induced contractions of the isolated guinea pig ileum were minimally enhanced or suppressed by very high concentrations (10(-4) g/ml) of SA-103 and DSCG. In addition to the above results, prolonged treatment with either compound before antigen challenge decreased the inhibitory response to anaphylactic histamine release from rat peritoneal cells. It is suggested, therefore, that the main mechanism(s) of the anti-allergic action of SA-103 is similar to that of DSCG, and SA-103 may be expected to be effective against allergic diseases.