Yasunari Nakata

Expression of brain specific chondroitin sulfate proteoglycans, neurocan and phosphacan, in the developing and adult hippocampus of Ihara's epileptic rats.

Iharas epileptic rats (IER) is an animal model of temporal lobe epilepsy with mycrodysgenesis, that exhibit abnormal migration of hippocampal neurons and recurrent spontaneous seizures. As an attempt to elucidate the roles of extracellular matrix molecules in the epileptogenecity and mossy fiber sprouting, immunohistochemical localization of brain specific chondroitin sulfate proteoglycans (CSPGs), neurocan and phosphacan, was examined in the hippocampus of postnatal IER and Sprague-Dawley (SD) rats using monoclonal antibodies 1G2 against neurocan and 6B4 against phosphacan. There was no difference in the expression of these two CSPGs between IER and SD rats in the 1st postnatal week. However, the expression of neurocan was poor in the hippocampus of IER in the 2nd and 3rd weeks whereas intense labeling of neurocan was present throughout the hippocampus of SD rats. Labeling of neurocan was almost absent in the hippocampus, while phosphacan was diffusely expressed in the stratum oriens and radiatum of Ammons horn, and in the hilus and inner one-third molecular layer of the dentate gyrus at the 2nd month after birth. There was no difference in the expression of neurocan and phosphacan between IER and SD rats at the 2nd month after birth. By contrast, phosphacan was reduced in the inner molecular layer of the dentate gyrus in 8-month-old IER, while neurocan was reexpressed in the outer molecular layer and hilus in 3- and 8-month-old IER. It was suggested that the insufficient expression of neurocan may affect the development of neuronal organization in the hippocampus, and that the remodeling of extracellular matrix in the dentate gyrus may contribute to the mossy fiber sprouting into the inner molecular layer.

Simultaneous induction of matrix metalloproteinase-9 and interleukin 8 by all-trans retinoic acid in human PL-21 and NB4 myeloid leukaemia cells

Summary. All‐trans retinoic acid (ATRA) has been shown to induce differentiation of human acute promyelocytic leukaemia (APL) cells and eventual elimination of the malignant clone. Matrix metalloproteinase‐9 (MMP‐9) is produced by neutrophils and its expression appears to be linked with myeloid cell differentiation. We investigated effects of ATRA on MMP expression in two human myeloid leukaemia cell lines, PL‐21 and NB4. Both cells could differentiate into neutrophils after exposure to ATRA. Both the activity and antigen levels of MMP‐9 were much higher in NB4 cells than in PL‐21 cells. Stimulation with ATRA significantly increased MMP‐9 levels approximately three‐ to fivefold in both PL‐21 and NB4‐conditioned media. MMP‐9 mRNA levels increased in ATRA‐treated cells and was almost in parallel with the increase in MMP‐9 activity, suggesting that ATRA induced MMP‐9 by activating its gene expression. ATRA can induce interleukin 8 (IL‐8) in APL cells. IL‐8, chemokine for neutrophils and a potent inducer of MMP‐9, was also induced by ATRA in PL‐21 cells. However, recombinant IL‐8 did not induce MMP‐9 expression. In addition, a neutralizing antibody against IL‐8 did not inhibit ATRA‐induced MMP‐9 expression in either cell type. These observations suggest that ATRA can induce both MMP‐9 and IL‐8, but IL‐8 is not involved in ATRA‐induced MMP‐9 expression. As MMP‐9 can truncate and activate IL‐8, simultaneous induction of MMP‐9 and IL‐8 by ATRA could activate leucocytes excessively, causing the hyper‐inflammatory events in retinoic acid syndrome.

Sarcoidosis occurring after interferon-alpha therapy for chronic hepatitis C: report of two cases.

Abstract:  We report two patients who were diagnosed with sarcoidosis after receiving interferon (IFN)‐α therapy for chronic hepatitis C, and conduct a review the relevant literature. The first patient was a 52‐year‐old female who developed multiple subcutaneous nodules 2 months after finishing IFN‐α therapy. A skin biopsy from subcutaneous nodules on the right elbow joint revealed sarcoid granulomata. These lesions resolved spontaneously 4 months later. The second patient, a 57‐year‐old male, developed bilateral hilar and mediastinal lymph node enlargement 2 years after finishing IFN‐α 2a therapy. A transbronchial lung biopsy demonstrated sarcoid granulomata. In addition, he had uveitis and left ulnar nerve involvement. His eye and nerve involvement gradually improved over 20 months. It is feasible that IFN therapy has been a trigger for sarcoidosis in these patients.

Induction of IL-8 and monoclyte chemoattractant protein-1 by doxorubicin in human small cell lung carcinoma cells

We previously demonstrated doxorubicin‐induced urokinase expression in human H69 SCLC cells by the microarray technique using Human Cancer CHIP version 2 (Takara Shuzo, Kyoto, Japan), in which 425 human cancer‐related genes were spotted on glass plates (Kiguchi et al., Int J Cancer 2001;93:792–7). Microarray analysis also revealed significant induction of IL‐8, a member of the CXC chemokines. We have, therefore, extended the observation by testing the effects of doxorubicin on expression of the chemokine family and provide here definitive evidence that doxorubicin induces IL‐8 and MCP‐1, one of the CC chemokines, at least in 2 human SCLC cells, H69 and SBC‐1. IL‐8 antigen levels, measured by ELISA, were markedly increased in both H69 and SBC‐1 conditioned media after doxorubicin treatment, in parallel with mRNA levels; and this was dependent on the dose of doxorubicin. The ribonuclease protection assay, using a multiprobe template set for human chemokines, revealed induction of not only IL‐8 but also MCP‐1 in doxorubicin‐treated H69 cells. MCP‐1 antigen levels increased approximately 100‐fold in doxorubicin‐treated H69 cells. RT‐PCR using specific primers for MCP‐1 suggested that doxorubicin also induced MCP‐1 expression in SBC‐1 and SBC‐3 SCLC cells. Futhermore, CAT analysis using IL‐8 promoter implicated the PEA3 transcriptional factor, whose binding site was located immediately upstream of the AP‐1 and NF‐κB binding sites. Thus, it is suggested that doxorubicin induces IL‐8 and MCP‐1 chemokines in human SCLC cells by activating gene expression, in which at least PEA3 is involved. IL‐8 and MCP‐1 are major chemoattractants for neutrophils and monocytes/macrophages, respectively; therefore, extensive induction of IL‐8 and MCP‐1 may provoke the interaction between inflammatory/immune cells and tumor cells under doxorubicin stimulation and influence many aspects of tumor cell biology.

Developmentally regulated expression of brain-specific chondroitin sulfate proteoglycans, neurocan and phosphacan, in the postnatal rat hippocampus.

Abstract. Developmental changes in the distribution of brain-specific chondroitin sulfate proteoglycans, neurocan and phosphacan/RPTPζ/β, in the hippocampus of the Sprague-Dawley rat were examined using monoclonal antibodies 1G2 and 6B4. The 1G2 immunoreactivity was predominant in the neonatal hippocampus while the 6B4 immunoreactivity was predominant in the mature hippocampus. Moderate 1G2 immunoreactivity was detected in the dentate gyrus and subiculum immediately after birth. Immunoreactivity reached a peak on postnatal days 7–10 (P7–P10) when intense 1G2 labeling was present throughout the neuropil layers of the hippocampus except the mossy fiber tract. 6B4 immunoreactivity was limited in the stratum lacunosum moleculare of CA1 in the neonatal hippocampus. It gradually increased by P21 when diffuse 6B4 immunoreactivity was detected in the stratum oriens and radiatum of Ammons horn, and in the hilus and inner one-third molecular layer of the dentate gyrus, while 1G2 immunoreactivity decreased after P21. In the adult hippocampus, moderate 6B4 immunoreactivity was present in the stratum oriens and radiatum of Ammons horn, and in the hilus and inner one-third molecular layer of the dentate gyrus, but not in the mossy fiber tract. In addition, strong 6B4 labeling appeared around a subset of neurons after P21. The results suggest that neurocan may have a role in the development of neuronal organization, while phosphacan/RPTPζ/β may contribute to the maintenance and plasticity of synaptic structure and function. Furthermore, the absence of 1G2 and 6B4 immunoreactivities in the stratum lucidum suggests that neurocan and phosphacan/RPTPζ/β may function as a barrier for the extension of mossy fibers and provide an environment permissive for fasciculation of the mossy fibers.

Induction of urokinase‐type plasminogen activator by the anthracycline antibiotic in human RC‐K8 lymphoma and H69 lung‐carcinoma cells

Current evidence has suggested the possible involvement of ROS as signaling messengers in IL‐1β‐ or LPS‐induced gene expression. We previously reported that both IL‐1β and LPS induce uPA in RC‐K8 human lymphoma cells. Here, we provide evidence that ROS‐generating anthracycline antibiotics, including doxorubicin and aclarubicin, upregulate uPA expression in 2 human malignant cell lines, RC‐K8 and H69 small‐cell lung‐carcinoma cells. Both doxorubicin and aclarubicin markedly increased uPA accumulation in RC‐K8‐ and H69‐conditioned medium in a dose‐dependent manner. In each case, maximal induction was observed at a sublethal concentration, i.e., at a concentration where cell growth was slightly inhibited. Both doxorubicin and aclarubicin increased uPA mRNA levels, and induction in each case reached the maximal level 9 hr after stimulation. Doxorubicin barely changed the half‐life of uPA mRNA and activated uPA gene transcription. Antioxidants such as NAC and PDTC inhibited doxorubicin‐induced uPA mRNA accumulation. Microarray analysis, using Human Cancer CHIP version 2 (Takara Shuzo, Kyoto, Japan), in which 425 human cancer‐related genes were spotted on glass plates, revealed that uPA is 1 of 3 genes that were clearly upregulated in H69 cells by doxorubicin stimulation. These findings suggest that the anthracycline induces uPA in human malignant cells by activating gene transcription in which ROS may be involved. Therefore, by upregulating uPA expression, the anthracycline may influence many biologic cell functions mediated by the uPA/plasmin system.

First presenting signs or symptoms of sarcoidosis in a Japanese population.

PurposeTo determine the first presenting signs or symptoms or other reasons leading to the diagnosis of sarcoidosis.MethodsA retrospective review was made of the records of 123 consecutive Japanese patients with histopathological diagnosis of sarcoidosis seen at a referral-based university hospital.ResultsAt the first presentation, eye symptoms in 32 patients, abnormal chest X-ray findings in 52 patients, common cold-like symptoms in 12 patients, lymphadenopathy in 6 patients, skin lesions in 14 patients, and examinations for other diseases in 4 patients led to the final diagnosis. Overall, uveitis was detected in 60 patients (50%) during the follow-up.ConclusionsMass screening programs of chest X-rays are the major way sarcoidosis is detected in Japan. Uveitis is seen in about half the patients during the course of sarcoidosis, and eye symptoms are frequent first presentations of sarcoidosis. These facts emphasize the role of ophthalmologists in the diagnosis and management of sarcoidosis.

Decreased monocyte-mediated cytostasis of human cancer cell in patients with lung cancer

SummaryIn vivo animal studies support the concept that monocytes and macrophages are important in the immune surveillance of oncogenesis and that in vitro activated murine macrophages are cytocidal for tumour cells. In this study, the tumour cell cytotoxic activity of human peripheral blood monocytes was examined by measuring the inhibition of 3H-thymidine uptake in the human cancer cell line, established in our laboratory from human squamous cell lung cancer. The monocytes from 8 of the 31 lung cancer patients (26%) showed a percentage growth inhibition of less than 69.8%, which exceeded the 95% confidence limits of the percentage growth inhibition observed with healthy control monocytes. On the other hand, among the 16 sarcoidosis and the 8 tuberculosis cases no value was below 69.8%. However, there was no significant difference between the growth inhibition and the clinical stages or histological type. When OK-432, a Streptococal agent, was administered in vivo to patients with lung cancer, an elevation of the growth inhibition was observed in 7 out of 8 patients. It was confirmed that the tumour cell cytostatic activity of the monocyte is suppressed in patients with lung cancer, and these monocyte deficits hinder the inhibition of tumour growth and metastasis.

EFFECT OF BESTATIN ON BONE MARROW

Publisher Summary This chapter reviews the effect of bestatin on bone marrow. In a study described in the chapter, bone marrow cells were aspirated from the sternum using Komiyas bone marrow aspiration needle. The administration of bestatin increased the nuclear cell count of bone marrow in six of the seven patients, and it also increased the mean value from 69, 400 ± 25,500/cmm to 99,500 ± 50,000/cmm. The absolute count of each cell was obtained by multiplying the nuclear cell count by the percentage of neutrophilic series or that of the erythrocytic series, respectively. In all the seven cases, the administration of bestatin increased the count of the neutrophilic series of bone marrow. The mean value before administration was 33, 200 ± 18,300/cmm and that after administration was 56, 200 ± 28,200/cmm. An increase in the count of the erythrocytic series of bone marrow was observed in four of seven patients. The peripheral blood picture at the time of bone marrow aspiration was examined. An increase in the WBC count was observed in six of seven patients, and an increase in neutrophil count was observed in all seven patients; no change in the hemoglobin content was observed.

Myocardial Sarcoidosis With A Tumor-Like Left Atrial Thrombus

We report here an elderly woman who started vague complaints around the age of 50, was proved to be a sarcoidosis by negative skin reaction with purified protein derivative, bilateral hilar lymphadenopathy, and sarcoid lesions in biopsied liver and lymph nodes, and died of cardiac insufficiency after 15 years of the illness. Necropsy revealed a huge tumor‐ like left atrial thrombus with nonspecific fibrous lesions throughout the myocardium, a pulmonary hamartoma, pneumonia, liver cell necrosis, and cholecystopathy. To our knowledge, this may be the first case of myocardial sarcoidosis associated with this kind of atrial thrombus, although the sarcoidosis and thrombus may have occurred independently.