BACKGROUND: The present investigation was undertaken to evaluate the neuroprotective effect of etomidate against kainic acid (KA) induced neurotoxicity in rats by using the immunoreactivity of heat shock protein-70 (HSP-70) and the acid-fuchsin stain. METHODS: Administration of etomidate (20 mg/kg, I.P.) was performed in sequence; first being just one hour after a KA (10 mg/kg, I.P.) injection, then three more times at one hour intervals. Neuronal damages in the hippocampus were evaluated by using the acid-fuchsin stain to detect cell death and HSP-70 induction as an index of cell injury at 24 h after the administration of KA. RESULTS: HSP-70 induction and acid fuchsin positive neurons were increased in the CA1 and CA3 regions of the hippocampus after a KA injection but significantly decreased by an injection of etomidate (P < 0.01). CONCLUSIONS: These results suggest that the etomidate has a potential effect on the protection of neurons against KA-induced neurotoxicity.
Animals ; Cell Death ; Etomidate* ; Hippocampus* ; Hot Temperature ; Kainic Acid ; Neurons ; Neuroprotective Agents ; Rats* ; Rosaniline Dyes ; Shock

Two cases of incomplete transverse vaginal septum with a small opening in the upper third of vagina were presented. One case was a patient presenting at 31 years of age for evaluation of primary infertility and oligomenorrhea. Diagnosis was made by hysterosalpingography, tranvaginal ultrasonography, MRI, and diagnostic pelviscopy for evaluation of abnormality of endopelvic organs. The patient was treated with surgical resection of septum (simple septectomy). Examination 3 months later demonstrated narrowing of the vaginal lumen. Another case was a patient presenting intrauterine pregnancy at 8 weeks and 2 days accompanying transverse vaginal septum with small opening. She had previous septotomy history and presented postoperative scarring. She had septectomy and anastomosis of the mucosa, and presented postoperative narrowing again about a month postoperatively. In this report we describe the two cases of transverse vaginal septum. We also reviewed the literature including diagnostic and treatment guidelines. Prevention of postoperative scarring is the most important factor in management of transverse vaginal septum. For that, first of all, initial septectomy should be done with approximation of the mucosa and postoperative dilatation is necessary. Importance of the initial accurate septectomy is too important to be over-emphasized.
Cicatrix ; Constriction, Pathologic* ; Diagnosis ; Dilatation ; Female ; Humans ; Hysterosalpingography ; Infertility ; Magnetic Resonance Imaging ; Mucous Membrane ; Oligomenorrhea ; Pregnancy ; Ultrasonography ; Vagina

In light of the anti-inflammatory properties of histone deacetylase (HDAC) inhibitors, such as suberoylanilide hydroxamic acid (SAHA) and trichostatin A (TSA), we examined a new HDAC inhibitor KBH-A42 for its anti-inflammatory activities. KBH-A42 showed noteworthy anti-inflammatory properties in vitro via suppression of the production of TNF-alpha, a proinflammatory cytokine, and nitric oxide (NO), a proinflammatory effector molecule, in LPS-stimulated RAW264.7 cells and peritoneal macrophages. It also inhibited TNF-alpha production in vivo as demonstrated in a LPS-induced mouse endotoxemia model. The levels of TNF-alpha, IL-1beta, IL-6 and iNOS mRNAs determined by RT-PCR propose that the inhibition of these pro-inflammatory mediators by KBH-A42 resulted from inhibiting expression of these genes. However, the EMSA study to see the effect of KBH-A42 on the binding of NF-kappaB, a transcription factor, to a specific DNA sequence showed that the binding of NF-kappaB to DNA was not changed regardless of increasing the concentration of KBH-A42 in the presence and absence of LPS stimulation. Interestingly, DNA binding of another transcription factor AP-1 dose-dependently increased by KBH-A42. KBH-A42 differentially regulated the phosphorylation of MAP kinases. While the phosphprylation of ERK1/2 and SAPK/JNK was not affected by KBH-A42, the phosphorylation of p38 decreased by KBH-A42. These results showed that KBH-A42 inhibits production of proinflammatory cytokines in macrophages by decreasing their mRNA levels, and p38 kinase is involved in the KBH-A42-mediated inhibition.
Animals ; Blotting, Western ; Cell Line ; Cell Survival/drug effects ; Cytokines/blood/genetics/*metabolism ; Electrophoretic Mobility Shift Assay ; Endotoxemia/blood/metabolism/pathology ; Enzyme Inhibitors/chemistry/*pharmacology ; Histone Deacetylases/*antagonists & inhibitors ; Hydroxamic Acids/chemistry/*pharmacology ; Interleukin-1beta/genetics/metabolism ; Interleukin-6/genetics/metabolism ; Macrophages/cytology/*drug effects/metabolism ; Mice ; Mitogen-Activated Protein Kinase 1/metabolism ; Mitogen-Activated Protein Kinase 3/metabolism ; Mitogen-Activated Protein Kinases/metabolism ; Molecular Structure ; NF-kappa B/metabolism ; Nitric Oxide/metabolism ; Nitric Oxide Synthase Type II/genetics/metabolism ; Phosphorylation/drug effects ; Piperidones/chemistry/*pharmacology ; Protein Binding/drug effects ; Reverse Transcriptase Polymerase Chain Reaction ; Transcription Factor AP-1/metabolism ; Tumor Necrosis Factor-alpha/blood/genetics/metabolism

Malignant rhabdoid tumors were first described in 1978 by Beckwith and Palmer as a rare variant of Wilms' tumors with a "rhabdomyosarcomatoid" pattern and a particularly poor prognosis. Week reclassified this disease as a distinct disease in 1989 and thereafter, there have been several reports about malignant rhabdoid tumor that occurred in various organs, including the colon. The histologic characteristics of rhabdoid tumor are noncohesive or loosely cohesive cells with high cellularity, an eccentric large nucleus and eosinophilic cytoplasm, and the cytoplasm is usually positive for vimentin and it contain hyaline inclusions. On immunohistochemical staining, the cells are usually positive for vimentin and cytokeratin and they are negative for desmin. This tumor progresses rapidly and it has a very poor prognosis, but survival is better if there is no lymphatic or distant metastasis. We experienced a patient who suffered with undifferentiated adenocarcinoma with rhabdoid features in the ascending colon.
Adenocarcinoma ; Colon ; Colon, Ascending ; Colonic Neoplasms ; Cytoplasm ; Desmin ; Eosinophils ; Humans ; Hyalin ; Keratins ; Neoplasm Metastasis ; Prognosis ; Rhabdoid Tumor ; Vimentin ; Wilms Tumor

Replication-independent incorporation of variant histone H3.3 has a profound impact on chromatin function and numerous cellular processes, including the differentiation of muscle cells. The histone chaperone HIRA and H3.3 have essential roles in MyoD regulation during myoblast differentiation. However, the precise mechanism that determines the onset of H3.3 deposition in response to differentiation signals is unclear. Here we show that HIRA is phosphorylated by Akt kinase, an important signaling modulator in muscle cells. By generating a phosphospecific antibody, we found that a significant amount of HIRA was phosphorylated in myoblasts. The phosphorylation level of HIRA and the occupancy of phosphorylated protein on muscle genes gradually decreased during cellular differentiation. Remarkably, the forced expression of the phosphomimic form of HIRA resulted in reduced H3.3 deposition and suppressed the activation of muscle genes in myotubes. Our data show that HIRA phosphorylation limits the expression of myogenic genes, while the dephosphorylation of HIRA is required for proficient H3.3 deposition and gene activation, demonstrating that the phosphorylation switch is exploited to modulate HIRA/H3.3-mediated muscle gene regulation during myogenesis.
Antibodies, Phospho-Specific ; Chromatin ; Histones* ; Muscle Cells* ; Muscle Development ; Muscle Fibers, Skeletal ; Myoblasts ; Phosphorylation* ; Phosphotransferases ; Transcriptional Activation

Various cell types in higher multicellular organisms are genetically homogenous, but are functionally and morphologically heterogeneous due to the differential expression of genes during development, which appears to be controlled by epigenetic mechanisms. However, the exact molecular mechanisms that govern the tissue-specific gene expression are poorly understood. Here, we show that dynamic changes in histone modifications and DNA methylation in the upstream coding region of a gene containing the transcription initiation site determine the tissue-specific gene expression pattern. The tissue-specific expression of the transgene correlated with DNA demethylation at specific CpG sites as well as significant changes in histone modifications from a low ratio of methylated H3- lysine 4 or acetylated H3-lysine 9, 14 to acetylated H4 to higher ratios. Based on the programmed status of transgene silenced in cloned mammalian ear-derived fibroblasts, the transgene could be reprogrammed by change of histone modification and DNA methylation by inhibiting both histone deacetylase and DNA methylation, resulting in high expression of the transgene. These findings indicate that dynamic change of histone modification and DNA methylation is potentially important in the establishment and maintenance of tissue-specific gene expression.
Transgenes/*genetics ; Swine ; Organ Specificity/genetics ; Methylation ; Lysine/*metabolism ; Histones/*metabolism ; Histone Deacetylases/metabolism ; Gene Silencing ; *Gene Expression ; Fibroblasts ; Ear ; *DNA Methylation ; Cells, Cultured ; Animals, Genetically Modified ; Animals ; Acetylation

Lysophosphatidic acid (LPA) is a bioactive phospholipids and involves in various cellular events, including tumor cell migration. In the present study, we investigated LPA receptor and its transactivation to EGFR for cyclooxygenase-2 (COX-2) expression and cell migration in CAOV-3 ovarian cancer cells. LPA induced COX-2 expression in a dose-dependent manner, and pretreatment of the cells with pharmacological inhibitors of Gi (pertussis toxin), Src (PP2), EGF receptor (EGFR) (AG1478), ERK (PD98059) significantly inhibited LPA- induced COX-2 expression. Consistent to these results, transfection of the cells with selective Src siRNA attenuated COX-2 expression by LPA. LPA stimulated CAOV-3 cell migration that was abrogated by pharmacological inhibitors and antibody of EP2. Higher expression of LPA2 mRNA was observed in CAOV-3 cells, and transfection of the cells with a selective LPA2 siRNA significantly inhibited LPA-induced activation of EGFR and ERK, as well as COX-2 expression. Importantly, LPA2 siRNA also blocked LPA-induced ovarian cancer cell migration. Collectively, our results clearly show the significance of LPA2 and Gi/Src pathway for LPA-induced COX-2 expression and cell migration that could be a promising drug target for ovarian cancer cell metastasis.
Butadienes/pharmacology ; Cell Line, Tumor ; Cell Movement/drug effects/*physiology ; Cyclooxygenase 2/*biosynthesis ; Extracellular Signal-Regulated MAP Kinases/antagonists & inhibitors/metabolism ; Female ; Flavonoids/pharmacology ; GTP-Binding Protein alpha Subunits, Gi-Go/antagonists & inhibitors/*metabolism ; Humans ; Lysophospholipids/pharmacology ; Nitriles/pharmacology ; Ovarian Neoplasms/metabolism/*pathology ; Pertussis Toxin/pharmacology ; Protein-Tyrosine Kinases/antagonists & inhibitors/*metabolism ; Proto-Oncogene Proteins/antagonists & inhibitors/*metabolism ; Pyrimidines/pharmacology ; Receptor, Epidermal Growth Factor/antagonists & inhibitors/metabolism ; Receptors, Lysophosphatidic Acid/*metabolism ; Receptors, Prostaglandin E/metabolism ; Signal Transduction ; Transcriptional Activation ; Tyrphostins/pharmacology

Xanthohumol (XH), the principal prenylflavonoid of the hop plant (Humulus lupulus L.), dose-dependently inhibited isobutylmethylxanthine (IBMX)-induced melanogenesis in B16 melanoma cells, with little cytotoxicity at the effective concentrations. Decreased melanin content was accompanied by reduced tyrosinase enzyme activity, protein and mRNA expression. The levels of tyrosinase-related protein 1 and 2 mRNAs were decreased by XH. XH also inhibited alpha-melanocyte stimulating hormone- or forskolin-induced increases in melanogenesis, suggesting an action on the cAMP-dependent melanogenic pathway. XH downregulated the protein and mRNA expression of microphthalmia-associated transcription factor (MITF), a master transcriptional regulator of key melanogenic enzymes. These results suggest that XH might act as a hypo-pigmenting agent through the downregulation of MITF in the cAMP-dependent melanogenic pathway.
1-Methyl-3-isobutylxanthine/pharmacology ; Animals ; Cell Line ; Cell Survival/drug effects ; Dose-Response Relationship, Drug ; Down-Regulation ; Drug Antagonism ; Forskolin/pharmacology ; *Humulus ; Intramolecular Oxidoreductases/antagonists & inhibitors/biosynthesis ; Melanins/antagonists & inhibitors/*biosynthesis ; Melanocytes/*drug effects/*metabolism ; Melanoma, Experimental ; Membrane Glycoproteins/antagonists & inhibitors/biosynthesis ; Mice ; Microphthalmia-Associated Transcription Factor/antagonists & inhibitors ; Monophenol Monooxygenase/antagonists & inhibitors/biosynthesis/genetics ; Oxidoreductases/antagonists & inhibitors/biosynthesis ; Propiophenones/*pharmacology ; Signal Transduction/drug effects ; alpha-MSH/metabolism