Enhanced miR-210 expression promotes the pathogenesis of endometriosis through activation of signal transducer and activator of transcription 3.

STUDY QUESTION: What are the roles of the microRNA miR-210-an miRNA that is up-regulated in endometriotic cyst stromal cells (ECSCs)-in the pathogenesis of endometriosis? SUMMARY ANSWER: Up-regulated miR-210 expression in ECSCs is involved in their proliferation, resistance to apoptosis and angiogenesis through signal transducer and activator of transcription (STAT) 3. WHAT IS KNOWN ALREADY: In the pathogenesis of endometriosis, a number of roles for microRNAs (miRNAs) are becoming apparent. STUDY DESIGN, SIZE, DURATION: ECSCs and normal endometrial stromal cells (NESCs) were isolated from ovarian endometriotic tissues (patients aged 24-40 years undergoing salpingo-oophorectomy or evisceration for the treatment of ovarian endometriotic cysts, n = 10) and the eutopic endometrial tissues without endometriosis (premenopausal patients aged 35-45 years undergoing hysterectomies for subserousal leiomyoma, n = 13), respectively. PARTICIPANTS/MATERIALS, SETTING, METHODS: We used a global gene expression microarray technique to identify downstream targets of miR-210, and we assessed the functions of miR-210 in the pathogenesis of endometriosis by using the miR-210-transfected NESCs. MAIN RESULTS AND THE ROLE OF CHANCE: Gene expression microarray analysis revealed that one of the key target molecules of miR-210 is STAT3. In the NESCs, in comparison to the control, miR-210 transfection resulted in the induction of cell proliferation (P < 0.0005), the production of vascular endothelial cell growth factor (VEGF) (P < 0.0005) and the inhibition of apoptosis (P < 0.05) through STAT3 activation [increased levels of mRNA (P < 0.0005), and protein (P < 0.005)]. In the ECSCs, inhibitors of STAT3 inhibited the cell proliferation and VEGF production (P < 0.05), and induced the apoptosis of these cells (P < 0.05). LIMITATIONS, REASONS FOR CAUTION: The roles of aberrant miR-210 expression were investigated only in the stromal component of ectopic and eutopic endometrium. Control endometrial tissues were obtained from premenopausal patients who had subserosal leiomyoma and NESC gene expression patterns may be altered in these women. Furthermore, the effects of STAT3 inhibitors were evaluated only in ECSCs and not in NESCs. WIDER IMPLICATIONS OF THE FINDINGS: The present findings indicate that miR-210 induces NESCs to differentiate into the endometriotic phenotype and we speculate that up-regulated miR-210 expression in ECSCs is involved in the creation of the endometriosis-specific cellular dysfunctions through epigenetic mechanisms. The data indicate that STAT3 inhibitors may be promising candidates for the treatment of endometriosis. STUDY FUNDING/COMPETING INTERESTS: This work was supported in part by Grants-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (no. 13237327 to K.N., no. 25861500 to Y.K. and no. 23592407 to H.N.). There are no conflicts of interest to declare.

The role of serum adiponectin levels in women with polycystic ovarian syndrome.

PURPOSE OF INVESTIGATION: The aim of this study was to measure serum adiponectin concentrations in women with polycystic ovarian syndrome (PCOS) and to assess possible correlations between adiponectin and the hormonal or metabolic parameters of this syndrome. MATERIALS AND METHODS: Serum adiponectin levels were evaluated in 20 women with PCOS and 22 women without PCOS whose age and body mass index (BMI) matched the patients. The levels of fasting blood glucose, fasting insulin, gonadotropin, and sex steroid hormones were evaluated in both groups. The homeostasis model assessment (HOMA) score was also calculated. The serum adiponectin levels were assayed by enzyme-linked immunoabsorbent assay (ELISA). RESULTS: Serum adiponectin levels were significantly lower in obese women than in normal-weight women, and they were also significantly lower in PCOS patients with HOMA scores greater than 1.7 compared with those with HOMA scores lower than 1.7. When the subjects were divided in two groups based on serum adiponectin levels (> 40 microg/ml, < 40 microg/ml), 65% of patients with PCOS were included in the lower adiponectin group (p < 0.05). In addition, gonadotropin levels were increased, dependent on the adiponectin levels in women with PCOS. CONCLUSION: Adiponectin is regarded as a possible link between adiposity and insulin resistance (IR). From this data, the secretions of gonadotropin are implicated in the levels of adiponectin in women with PCOS. It is suggested that adiponectin may play an important role in the pathogenesis of PCOS.

Malignant lymphoma of the vagina successfully treated with rituximab, adryamicin, cyclophosphamide, vincristine sulfate, and prednisolone.

PURPOSE: Primary malignant lymphoma of the vagina is extremely rare. The most common histologic subtype is diffuse large B-cell lymphoma (DLBCL). We report a case of vaginal DLBCL successfully treated with chemotherapy consisting of rituximab, adryamicin, cyclophosphamide, vincristine sulfate, and prednisolone (R-CHOP), followed by pelvic irradiation. CASE: A 44-year-old Japanese woman was admitted complaining of atypical genital bleeding and puruloid vaginal discharge. Gynecological examination showed an ulceration of the vaginal wall and a hard mass the size of a goose egg beneath the left vaginal wall, which had infiltrated to the left pelvic wall. The pathological diagnosis based on a punch biopsy taken from the vaginal tumor was non-Hodgkin's lymphoma. Based on immunohistochemical study, the tumor was subclassified as activated B-cell type DLBCL. The patient was diagnosed with Ann Arbor Stage IEA DLBCL and Stage III vaginal cancer, according to the International Federation of Gynecologists and Obstetricians (FIGO) classification system. She was successfully treated by six courses of R-CHOP, followed by radiation therapy. The patient is well without evidence of disease 13 months following the initial treatment. CONCLUSION: Little attention has been paid to the use of rituximab in addition to conventional chemotherapy and the importance of clinical and morphological subgrouping of DLBCL arising in the vagina. The present case indicates that the effects of rituximab on the prognosis of vaginal DLBCL must be evaluated, and that clinical use of immunophenotypic subgrouping should be considered for vaginal DLBCL.

Regulation of interleukin-1alpha and tumor necrosis factor-alpha-induced interleukin-8 production by amnion-derived (WISH) cells.

PURPOSE OF INVESTIGATION: It has been reported that interleukin (IL)-8 is produced in the amnion and that its production is enhanced by the initiation of labor. The purpose of this study was to clarify the mechanism of IL-8 production by amnion-derived (WISH) cells. METHODS: Cells were cultured and treated with various concentrations of interleukin (IL)-1alpha, IL-1 receptor antagonist (ra), tumor necrosis factor (TNF)- alpha, C2-, C6-ceramide, mitogen-activated protein (MAP) kinase kinase (MEK) inhibitor (U0126) and pyridinyl imidazole (p38 MAP kinase inhibitor, SB203580). IL-8 in the culture medium was measured by ELISA. RESULTS: The production of IL-8 was significantly increased by IL-1alpha or TNF-alpha, and the increase of IL-8 stimulated by IL-1alpha was suppressed by IL-1 ra in a dose-dependent manner. The increase in IL-8 production by IL-1alpha or TNF-alpha was further enhanced by simultaneous addition of C2-ceramide. The increase of IL-8 stimulated by IL-1alpha or TNF-alpha was also suppressed by treatment with U0126 or SB203580. The results of this study demonstrate that the production of IL-8 induced by IL-1alpha and TNF-alpha is enhanced by C2-ceramide, and suppressed by MEK inhibitor or P38 MAP kinase inhibitor. CONCLUSION: The results suggest that ceramide-mediated accumulation and MAP kinase-mediated suppression of inflammatory events in the amnion may play an important role in the maintenance of pregnancy and initiation of labor.

Depression and pregnancy-associated death by suicide after spinal cord injury: a case report.

PURPOSE OF INVESTIGATION: To report a case of a pregnant woman with traumatic spinal cord injury complicated with a psychiatric disorder. CASE REPORT: A 24-year-old woman at 18 weeks of gestation was transferred to our hospital with a history of having jumped from a third-floor apartment patio. RESULT: A trauma survey showed no life-threatening hemorrhage, and fetal wellbeing was confirmed. Neurological examination showed complete loss of motor and sensory function in her lower extremities. Termination of pregnancy was advised and was achieved medically. Surgical intervention was performed to achieve stabilization of the spine and decompression of neural elements. After the operation, she was referred to a psychiatrist, and the administration of paroxetine, etizolam and flunitrazepam was begun. Four months after undergoing the abortion, she choked herself to death on her ward bed. CONCLUSION: Although it is rare, we should pay special attention to the substantial suicide risk of women who face severe spinal cord injury.

Villoglandular papillary adenocarcinoma of the uterine cervix diagnosed during pregnancy.

Villoglandular papillary adenocarcinoma (VPA) is a very rare subtype of adenocarcinoma of the uterine cervix but a well recognized variant of cervical adenocarcinoma with a favorable prognosis generally occurring in women of child-bearing age. Only five cases of VPA and pregnancy have been reported. Herein, we report a case of VPA diagnosed during pregnancy and this patient delivered a healthy baby. A successful pregnancy can be completed in patients with VPA without lymph-vascular invasion, when treated conservatively. This management is particularly desirable in young women to preserve reproductive capability.

Primary mucinous adenocarcinoma of the vagina.

PURPOSE: Primary mucinous adenocarcinoma of the vagina is a rare disease which is characterized by aggressiveness and poor prognosis because of its rapid growth and recurrence, its frequent distant metastases, and its relative resistance to conventional treatment modalities including surgery, radiotherapy, and chemotherapy. We report a case of advanced stage primary mucinous adenocarcinoma of the vagina that showed a highly aggressive course and resistance to combination chemotherapy with paclitaxel and carboplatin. CASE: A 46-year-old multigravid Japanese woman was admitted to our hospital to be treated for Stage IVb primary mucinous adenocarcinoma of the vagina. She had no history of in utero exposure to diethylstilbestrol. She was treated by two courses of neoadjuvant chemotherapy with tri-weekly paclitaxel and carboplatin, which were not effective. Subsequently, total pelvic exenteration with pelvic lymphadenectomy was performed. However, the disease progressed rapidly and the patient died five months after the initial treatment. CONCLUSION: Because of its rarity, little is known about the behavior of primary mucinous adenocarcinoma of the vagina. Additional data about patients with this rare tumor should be collected and analyzed in an attempt to elucidate its prognostic factors, characteristics, optimal treatment, and outcome.

Nitrate ions: Potentiation of increased permeability to sugar associated with muscle contraction.

Nitrate ions potentiate twitch tension and enhance the increase in permeability to sugar which occurs in electrically stimulated frog sartorius muscles. However, the potentiating effect of nitrate ions on permeability is not dependent upon an increase in twitch tension. The possible relation of changes in permeability to alterations of the concentration of calcium ions in the cell is discussed.

Primary malignant melanoma of the vagina.

Malignant melanoma originating in the vagina is considered extremely rare and has a very poor prognosis. We report a case of a 70-year-old woman with primary malignant melanoma of the vagina, and discuss the importance of prognostic factors and the efficacy of adjuvant chemotherapy.

Bowel obstruction due to endometriosis in the rectovaginal septum.

It is very rare that endometriotic lesions in the rectovaginal septum cause ileus. We report a case of bowel obstruction due to endometriotic lesions in the rectovaginal septum in a 22-year-old woman whose barium enema presented with apple-core-like findings. Diagnostic and treatment modalities were discussed. Preoperative and postoperative gonadotropin-releasing hormone analog and aromatase inhibitor therapy promote relief of clinical symptoms, a reduction of tumor volume and a better approach to radical surgery.

Selective loss of a plasma membrane protein associated with contraction of skeletal muscle.

Muscle proteins were labeled by incubating isolated frog sartorius muscles with [3H]- or [14C]phenylalanine. Sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis of plasma membrane fractions revealed a major protein band with an apparent molecular weight of approx. 96,000. Radioactivity in this band showed a clearly delineated decrease, relative to other bands, when previously labeled muscles were induced to contract either by electrical stimulation or by increasing the influx of Ca2+ from the incubation medium. It is postulated that a Ca2+-activated neutral protease may account for this decrease in labeled membrane protein.

Use of tritiated 3-O-methyl-D-glucose for studies of membrane transport. Caveat.

Tritiated 3-O-methyl-D-glucose has many useful attributes as a model substance for studies of the transport of glucose across cell membranes. However, preparations of high specific radioactivity can decompose within a few months, producing radioactive impurities that can cause a several-fold increase in the apparent rate of sugar transport. In our investigation radioactive contaminants entered frog skeletal muscle cells by free diffusion rather than by facilitated transport. Much of the contaminating radioactive material could be removed by evaporating the solvent and redissolving the sugar. Tritiated sugar samples that had a specific activity below 0.1 Ci/mmol remained stable and suitable for transport measurements after several years of storage at -20 degrees C. In order to evaluate the suitability of a given tritiated preparation of sugar for transport measurements, it is recommended that its behavior be compared with that of a stable reference standard of low specific activity.

Isolation of plasma membrane vesicles, derived from transverse tubules, by selective homogenization of subcellular fractions of frog skeletal muscle in isotonic media.

A new technique for isolating fragmented plasma membranes from skeletal muscle has been developed that is based on gentle mechanical disruption of selected homogenate fractions. (Na+ + K+)-stimulated, Mg2+-dependent ATPase was used as an enzymatic marker for the plasma membrane, Ca2+-stimulated, Mg2+-dependent ATPase as a marker for sarcoplasmic reticulum, and succinate dehydrogenase for mitochondria. Cell segments in an amber low-speed (800 x g) pellet of a frog muscle homogenate were disrupted by repeated gentle shearing with a Polytron homogenizer. Sarcoplasmic reticulum was released into the low-speed supernatant, whereas most of the plasma membrane marker remained in a white, fluffy layer of the sediment, which contained sarcolemma and myofibrils. Additional gentle shearing of the white low-speed sediment extracted plasma membranes in a form that required centrifugation at 100,000 x g for pelleting. This pellet, the fragmented plasma membrane fraction, had a relatively high specific activity of (Na+ + K+)-stimulated ATPase compared with the other fractions, but it had essentially no Ca2+-stimulated ATPase activity and only a small percentage of the succinate dehydrogenase activity of the homogenate. Experimental evidence suggests that the fragmented plasma membrane fraction is derived from delicate transverse tubules rather than from the thicker, basement membrane-coated sarcolemmal sheath of muscle cells. Electron microscopy showed small vesicles lined bu a single thin membrane. Hydroxyproline, a characteristic constituent of collagen and basememt membrane, could not be detected in this fraction.

Enhanced permeability to sugar associated with muscle contraction. Studies of the role of Ca++.

When contractures were induced in isolated frog sartorius muscles with 4 mM caffeine, there was an increase in permeability of the muscle cells to 3-methylglucose. This observation suggests that the changes in permeability to sugar that are known to occur in electrically stimulated muscles may not be intimately related to the depolarization phase of the tissue response. Contractures that were elicited by exposing the muscles to a high concentration of K+ were also associated with an increased permeability to sugar. As the concentration of 45Ca in the medium was raised, more 45Ca entered the muscles during potassium contractures, and the contractures lasted longer, in agreement with the observations of other investigators. There was also a greater change in permeability to sugar when potassium contractures were elicited in the presence of higher concentrations of Ca++. The possibility that the enhanced permeability to sugar may be related to changes in the intracellular concentration of Ca++ is discussed.

Secretion of platelet-activating factor acetylhydrolase by human decidual macrophages.

Platelet-activating factor (PAF) metabolism in the maternal-fetal decidual interface has been investigated. Human decidua was obtained from patients at term and not in labor after cesarean section. The cells were isolated by enzymic digestion, followed by Ficoll-Paque centrifugation, or were purified further by discontinuous Percoll density gradient centrifugation. Cell populations were analyzed by flow cytometry after labeling with macrophage-specific antibodies. Twenty-seven percent of the cells obtained after enzymic digestion and Ficoll-Paque density gradient centrifugation had macrophage surface markers. The decidual cell population secreted PAF-acetylhydrolase (PAF-AH) activity. The secreted PAF-AH was the plasma-type isozyme. Synthesis and secretion was inhibited by actinomycin-D or cycloheximide. The PAF-AH activity secreted into the culture medium correlated positively with the number of macrophages. Flow cytometric purification yielded a 96% macrophage marker-positive population. The macrophages were shown to be the only cell types of decidual tissue that secreted PAF-AH. Treatment with anti-CD14 monoclonal antibody and complement specifically blocked PAF-AH secretion by collagenase-dispersed cells. It is concluded that decidual macrophages produce and secrete PAF-AH of the plasma type, and it is suggested that these cells may play an important role in PAF metabolism during parturition.

The inhibitory effect of 1,25-dihydroxyvitamin D3 on the secretion of platelet-activating factor acetylhydrolase by human decidual macrophages.

The effects of 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] and 25-hydroxyvitamin D3 on the secretion of a platelet-activating factor (PAF)-inactivating enzyme, PAF-acetylhydrolase (PAF-AH), by decidual macrophage populations were examined. 1,25-(OH)2D3 inhibited PAF-AH secretion by either decidual cells or flow cytometrically purified decidual macrophages. 25-Hydroxyvitamin D3 also decreased the enzyme secretion but at higher concentrations than those required for 1,25-(OH)2D3. The 1,25-(OH)2D3-induced inhibition was partially blocked by protein kinase C (PKC) inhibitors, sphingosine and 1-(5-isoquinolinesulfonyl)-2-metylpiperazine (H-7). An intracellular calcium channel blocker, bis-(o-aminophenoxy)-ethane-N,N,N',N'-tetraacetic acid, tetra(acetoxymethyl)-ester (BAPTA/AM), also partially blocked the inhibition by 1,25-(OH)2D3, whereas extracellular calcium channel blockers, verapamil and nifedipine, failed to prevent the inhibition. H-7 and BAPTA/AM additively blocked the 1,25-(OH)2D3-induced inhibition. A PKC activator, 12-O-tetradecanoylphorbol 13-acetate, also decreased PAF-AH secretion. The decrease was abolished by sphingosine or H-7. It is suggested that 1,25-(OH)2D3 may increase the concentration of PAF in the decidua via its inhibitory effect on PAF-AH secretion by decidual macrophages and that the inhibitory action may be mediated by intracellular calcium and PKC-dependent signal transduction. PAF and 1,25-(OH)2D3 may, therefore, play a cooperative role in the regulation of PAF metabolism at the maternal-fetal interface.