Hiroki Yoshioka

Increased interleukin-6 levels in peritoneal fluid of infertile patients with active endometriosis.

OBJECTIVE Our purpose was to investigate the relationship between the levels of interleukin-6, interleukin-6 soluble receptor, and tumor necrosis factor-alpha in peritoneal fluid and the size and number of active red endometriotic lesions. STUDY DESIGN In a university hospital 39 women of reproductive age underwent either laparoscopy for infertility workup or laparoscopic surgery for ovarian chocolate cysts. Peritoneal fluid was collected by laparoscopy. Active lesions, such as red flamelike lesions, glandlike lesions, and red vesicles, were scored according to the revised American Fertility Society classification system according to the size and number of active lesions. Peritoneal fluid levels of interleukin-6, interleukin-6 soluble receptor, and tumor necrosis factor-alpha levels were determined by enzyme-linked immunosorbent assays. The relationship between peritoneal fluid concentrations of interleukin-6 and tumor necrosis factor-alpha and the score of active endometriosis was investigated. RESULTS Peritoneal fluid levels of interleukin-6 and tumor necrosis factor-alpha were significantly higher in patients with endometriosis compared with patients without endometriosis. The concentrations in patients with active endometriosis increased as the size and the number of active lesions increased. Cyclic variations in interleukin-6 concentrations were seen in peritoneal fluid from patients with endometriosis; the concentrations in the secretary phase were significantly higher than those in the proliferative phase. CONCLUSIONS Increased peritoneal fluid levels of interleukin-6 in patients with active red endometriosis may relate to endometriosis-associated infertility and to the pathogenesis of endometriosis.

Menstrual cycle–specific inhibition of the proliferation of endometrial stromal cells by interleukin 6 and its soluble receptor ☆ ☆☆

OBJECTIVE This study investigated the possible roles of interleukin 6 and soluble interleukin 6 receptor in the growth of endometrial and endometriotic cells. STUDY DESIGN Endometrial and endometriotic stromal cells were collected from the uterus or from ovarian chocolate cysts. We examined the effects of interleukin 6, soluble interleukin 6 receptor, and a combination of both factors on the proliferation of endometrial and endometriotic stromal cells. The action of sex steroids on the interleukin 6 regulation of the growth of stromal cells was also evaluated. The gene expressions of interleukin 6 receptor and glycoprotein 130 were examined in endometrial and endometriotic cells by reverse transcription-polymerase chain reaction. RESULTS Interleukin 6 had no effect on the growth of stromal cells in tissue from the proliferative phase. In contrast, the addition of concentrations of >/=100 pg/mL interleukin 6 induced significant inhibition of stromal cell proliferation in tissue from the secretory phase. Similarly, the addition of soluble interleukin 6 receptor caused significant suppression in the growth of endometrial stromal cells in tissue from the secretory phase but not the proliferative phase. On the other hand, stromal cells of endometriotic tissues were resistant to interleukin 6, showing no inhibitory response. Although the combination treatment did not affect the proliferation of stromal cells of the proliferative phase and of endometriotic tissues, 10 pg/mL interleukin 6 inhibited proliferation of stromal cells of the secretory phase in the presence of 1 ng/mL soluble interleukin 6 receptor. Treatment with estradiol and progesterone for 10 days newly induced the inhibitory response to interleukin 6 in the endometrial cells from the proliferative phase. Expressions of transcripts of interleukin 6 receptor and glycoprotein 130 were observed in the endometrial cells from the proliferative and secretory phases and in endometriotic cells. CONCLUSIONS Interleukin 6 may play a central role in regulation of the growth of endometrial cells as a mediator of endocrine action. Endometriotic cells may behave differently from their normal counterparts in terms of the inhibitory regulation exerted by interleukin 6.

Nonsteroidal Anti-inflammatory Drug-activated Gene (NAG-1/GDF15) Expression Is Increased by the Histone Deacetylase Inhibitor Trichostatin A

Nonsteroidal anti-inflammatory drug-activated gene (NAG-1) is a putative tumor suppressor whose expression can be increased by drug treatment. Glioblastoma is the most common central nervous system tumor, is associated with high morbidity and mortality, and responds poorly to surgical, chemical, and radiation therapy. The histone deacetylase inhibitors are under current consideration as therapeutic agents in treating glioblastoma. We investigated whether trichostatin A (TSA) would alter the expression of NAG-1 in glioblastoma cells. The DNA demethylating agent 5-aza-dC did not increase NAG-1 expression, but TSA up-regulated NAG-1 expression and acted synergistically with 5-aza-dC to induce NAG-1 expression. TSA indirectly increases NAG-1 promoter activity and increases NAG-1 mRNA and protein expression in the T98G human glioblastoma cell line. TSA also increases the expression of transcription factors Sp-1 and Egr-1. Small interfering RNA experiments link NAG-1 expression to apoptosis induced by TSA. Reporter gene assays, specific inhibition by small interfering RNA transfections, and chromatin immunoprecipitation assays indicate that Egr-1 and Sp-1 mediate TSA-induced NAG-1 expression. TSA also increases the stability of NAG-1 mRNA. TSA-induced NAG-1 expression involves multiple mechanisms at the transcriptional and post-transcriptional levels.

Laparoscopic management of early primary abdominal pregnancy

Background Abdominal pregnancy is a very rare condition with a high mortality rate. Case A 29-year-old woman with a history of primary infertility was admitted because of lower abdominal pain, bloody vaginal discharge, and positive urine pregnancy test. Transvaginal ultrasonography revealed a 27 × 25-mm mass containing a gestational sac-like structure located outside the uterus. At laparoscopy, a clot was revealed including chorionic villi, adherent to the peritoneum in the right vesicouterine pouch. This was removed along with the adherent peritoneum. Postoperative histology reveled invasion of chorionic tissues into peritoneum. Conclusion Early diagnosis of an ectopic pregnancy by transvaginal ultrasonography enabled the laparoscopic management of early abdominal pregnancy.

DNA methylation-mediated silencing of nonsteroidal anti-inflammatory drug-activated gene (NAG-1/GDF15) in glioma cell lines.

Nonsteroidal anti‐inflammatory drug‐activated gene, NAG‐1, a transforming growth factor‐β member, is involved in tumor progression and development. The association between NAG‐1 expression and development and progression of glioma has not been well defined. Glioblastoma cell lines have lower basal expression of NAG‐1 than other gliomas and normal astrocytes. Most primary human gliomas have very low levels of NAG‐1 expression. NAG‐1 basal expression appeared to inversely correlate with tumor grade in glioma. Aberrant promoter hypermethylation is a common mechanism for silencing of tumor suppressor genes in cancer cells. In glioblastoma cell lines, NAG‐1 expression was increased by the demethylating agent, 5‐aza‐2′‐deoxycytidine. To investigate whether the NAG‐1 gene was silenced by hypermethylation in glioblastoma, we examined DNA methylation status using genomic bisulfite sequencing. The NAG‐1 promoter was densely methylated in several glioblastoma cell lines as well as in primary oligodendroglioma tumor samples, which have low basal expression of NAG‐1. DNA methylation at two specific sites (−53 and +55 CpG sites) in the NAG‐1 promoter was strongly associated with low NAG‐1 expression. The methylation of the NAG‐1 promoter at the −53 site blocks Egr‐1 binding and thereby suppresses Nag‐1 induction. Treatment of cells with low basal NAG‐1 expression with NAG‐1 inducer also did not increase NAG‐1. Incubation with a demethylation chemical increased Nag‐1 basal expression and subsequent incubation with a NAG‐1 inducer increased NAG‐1 expression. We concluded from these data that methylation of specific promoter sequences causes transcriptional silencing of the NAG‐1 locus in glioma and may ultimately contribute to tumor progression.

Proteasome inhibitor MG132 induces NAG-1/GDF15 expression through the p38 MAPK pathway in glioblastoma cells.

The expression of nonsteroidal anti-inflammatory drug-activated gene-1 (NAG-1) is regulated by the p53 and Egr-1 tumor suppressor pathways. Many anti-cancer drugs and chemicals induce NAG-1 expression, but the mechanisms are not fully understood. Transgenic mice expressing human NAG-1 are resistant to intestinal and prostate cancer, suggesting that NAG-1 is a tumor suppressor. Proteasome inhibitors exhibit anti-glioblastoma activities in preclinical studies. Here, we show that the proteasome inhibitors MG132 and bortezomib induced NAG-1 expression and secretion in glioblastoma cells. MG132 increased NAG-1 expression through transcriptional and post-transcriptional mechanisms. At the transcriptional level, the induction of NAG-1 required the -133 to +41 bp region of the promoter. At post-transcriptional levels, MG132 stabilized NAG-1 mRNA by increasing the half-life from 1.5 h to >8 h. Because of the dramatic increase in mRNA stability, this is likely the major contributor to MG132-mediated NAG-1 induction. Further probing into the mechanism revealed that MG132 increased phosphorylation of the p38 MAPK pathway. Consequently, inhibiting p38 phosphorylation blocked activation of the NAG-1 promoter and decreased mRNA stability, indicating that p38 MAPK activation mediates both MG132-dependent promoter activation and mRNA stabilization of NAG-1. We propose that the induction of NAG-1 by p38 MAPK is a potential contributor to the anti-glioblastoma activity of proteasome inhibitors.

The Cyclooxygenase Inhibitor Sulindac Sulfide Inhibits EP4 Expression and Suppresses the Growth of Glioblastoma Cells

EP4 expression in human glioblastoma cells correlates with growth on soft agar. The cyclooxygenase inhibitor sulindac sulfide first altered specificity protein-1 (Sp-1) and early growth response gene-1 expression, then increased the expression of nonsteroidal anti-inflammatory drug-activated gene 1 and activating transcription factor 3, and then decreased EP4 expression. EP4 suppression was dependent on blocking the Sp-1 binding sites in the human EP4 promoter. Mutation in the Sp-1 sites in EP4 altered the promoter activity and abolished sulindac sulfide effects. The inhibitory effect of sulindac sulfide on EP4 expression was reversed by PD98059, a mitogen-activated protein/extracellular signal–regulated kinase kinase-1/extracellular signal–regulated kinase inhibitor. Sp-1 phosphorylation was dependent on sulindac sulfide–induced Erk activation. Chromatin immunoprecipitation assay confirmed that Sp-1 phosphorylation decreases Sp-1 binding to DNA and leads to the suppression of EP4. Inhibition of cell growth on soft agar assay was found to be a highly complex process and seems to require not only the inhibition of cyclooxygenase activity but also increased expression of nonsteroidal anti-inflammatory drug-activated gene 1 and activating transcription factor 3 and suppression of EP4 expression. Our data suggest that the suppression of EP4 expression by sulindac sulfide represents a new mechanism for understanding the tumor suppressor activity.

Autocrine Effects of Transforming Growth Factor-α on the Development of Preimplantation Mouse Embryos

Purpose:We wished to explore the role of transforming growth factor (TGF)-α in mouse embryonic development.Methods:We examined the gene expression of TGF-α and epidermal growth factor receptor (EGFR) in mouse blastocysts by the reverse transcription–polymerase chain reaction and evaluated the effects of TGF-α on the development of preimplantation mouse embryos using TGF-α antisense oligodeoxynucleotide. Mouse teratocarcinoma F9 cells were also a subject of this study.Results:Gene transcripts of TGF-α and EGFR were present in both blastocysts and F9 cells. TGF-α significantly stimulated the rate of blastocoel expansion in early-cavitating blastocysts and the proliferation of F9 cells. Northern blot analysis showed that TGF-α gene expression in F9 cells was markedly suppressed in the presence of TGF-α antisense oligodeoxynucleotide. TGF-α antisense oligonucleotide significantly reduced the rate of blastocoel expansion and the growth of F9 cells. The inhibitory effects of TGF-α antisense oligonucleotide on blastocysts and F9 cells were reversed by the addition of TGF-α.Conclusions:The present observations suggest that TGF-α acts as an autocrine factor in the development of preimplantation mouse embryos.

Vessel wall enhancement in the diagnosis and management of primary angiitis of the central nervous system in children.

We describe two cases of primary angiitis of the central nervous system in children (cPACNS) diagnosed by vessel wall contrast enhancement on magnetic resonance imaging (MRI). Both patients developed acute cerebral infarction after fever and malaise. In patient 1, a 7-month-old boy, MRI revealed extensive cerebral infarction in the right middle cerebral artery (MCA) area and stenosis at the M1 portion of the right MCA. Oral glucocorticoid therapy was initiated. Vessel wall enhancement was ameliorated 3months after onset, and stenosis was mostly restored. Patient 2, a 5-year-old boy, suffered from cerebral infarction in the left MCA area, and stenosis was identified in the left internal carotid artery, left MCA, and left posterior cerebral artery. Although vessel wall enhancement was reduced after glucocorticoid therapy, vessel wall enhancement of left MCA re-emerged, accompanied by increased erythrocyte sedimentation rate (ESR) and, decreased cerebral blood flow (CBF) in the affected hemisphere. Intravenous methylprednisolone therapy followed by oral glucocorticoid and mycophenolate mofetil resulted in resolution of these findings. Vessel wall enhancement is a promising finding in the diagnosis of cPACNS. Disease flares occur rarely in medium-to-large vessel cPACNS during dose tapering. Vessel wall enhancement, ESR, and CBF may be useful for the assessment of the activity of angiitis.

Primary Central Nervous System Lymphoma of the Cerebellopontine Angle That Initially Occurred as Neurolymphomatosis of the Acoustic Nerve

We report a rare case of a primary central nervous system lymphoma (PCNSL) of the cerebellopontine angle (CPA) with infiltration into the pyramidal tract that initially presented as neurolymphomatosis (NL) of the acoustic nerve. A 60-year-old male suffered from right-side deafness and was referred to an otolaryngologist. Magnetic resonance imaging (MRI) showed fusiform enlargement of the right acoustic nerve with a hyperintense signal on a T2-weighted image (T2WI) and with gadolinium (Gd) enhancement, without an evidence of parenchymal CNS involvement. Although he was treated with steroids, his symptoms deteriorated. MRI was performed again and showed the mass lesion at the right CPA with enhancement. In addition to this, a lesion with slightly high intensity on a T2WI with Gd enhancement was observed along the right pyramidal tract. Despite steroid pulse therapy, the lesion rapidly progressed. We performed a tumor biopsy, and the histological diagnosis was diffuse large B-cell lymphoma. Pelvic, abdominal, and chest computed tomography scans, gallium cintigraphy, and bone marrow biopsy failed to detect any other evidence of lymphomatous involvement of other organs. We attempted high-dose methotrexate therapy (3.5 g/m2). We found a discrepancy in the therapeutic effect between the CPA lesion and the infiltrated lesion along the pyramidal tract; the lesions were chemo-resistant and chemo-sensitive, respectively. After completion of the second courses of chemotherapy, we began radiotherapy (total dose: 36 Gy). Four months after radiotherapy, the CPA tumor completely disappeared. Thirty-three months after the biopsy, he is doing well with a normal daily life and no signs of recurrence.