OBJECTIVE: Leptin, a secreted protein of the Ob gene by white adipose tissue, plays an important role in the regulation of food intake and energy consumption in the brain and acts as a potential growth stimulator in normal and neoplastic breast cancer cells. However, a potential role of leptin as an endocrine regulator is unknown in ovarian cancer. In the present study, we investigated the expression of leptin and its receptors in various histologic types of ovarian cancer and immortalized ovarian cancer cell lines to find out potential effect of leptin on the cell growth and activation of ovarian cancer cell line. METHODS: The ovarian cancer tissues, serous (n=18), mucinous (n=15), clear cell (n=12) and endometrioid type (n=14), were used for immuno-histochemical staining for leptin (Ob) and leptin-receptors (Ob -R). Ovrian cancer cell lines (non-mucinous: SNU-8, OVCAR-3, and SKOV-3) and mucinous: MUC) were used for RT-PCR, Western blot analysis, and [H3] thymidine incorporation assay for the cell growth and activation of mitogen-activated protein kinase. RESULTS: Both long (Ob-Rb) and short (Ob-Rt) isoforms of leptin receptors are expressed in non-mucinous type of ovarian cancer tissues (serous, clear cell carcinoma and endometrioid cell carcinoma) and in non-mucinous ovarian cancer cell lines (SNU-8, OVCAR-3 and SKOV-3 cells). However, leptin and its receptors are not found in mucinous cancer cells and mucinous cancer cell line (MUC). In immunohistochemical staining, the immunreactive leptin is expressed on the nuclei of the stratified cuboidal-to-columnar epithelial cells whereas its receptor was sparsely expressed on the innermost epithelial cell clusters and cytoplasm in non-mucinous tumor. However, there are no immunoreactive leptin and its receptor expressions in the mucinous tumor. In addition, treatment with leptin resulted in the growth stimulation of ovarian cancer cell line, an activation of ERK 1/2 and inhibition of constitutive phosphorylation of p38 Mitogen-activated protein kinase (MAPK). CONCLUSION: Taken together, our data demonstrates preliminary that the expression of leptin and its receptor is different according to the cell types of the ovarian cancer. Also it canbe thought that leptin immunolocalized on the nuclei in non-mucinous type but not in mucinous possibly acts as a nuclear transcription factor. Further studies are necessary to validate whether leptin may be a potential regulator for ovarian cancer.
Adipose Tissue, White ; Blotting, Western ; Brain ; Breast Neoplasms ; Cell Line ; Cytoplasm ; Eating ; Epithelial Cells ; Leptin ; Mucins ; Neoplasms, Glandular and Epithelial ; Ovarian Neoplasms ; Phosphorylation ; Protein Isoforms ; Protein Kinases ; Receptors, Leptin ; Thymidine ; Transcription Factors

We report a case of awakening during general anesthesia due to a vaporizer malfunction. The sevoflurane vaporizer had a hole through which approximately 20% of fresh gas escaped. The gas in the common gas outlet contained only 60% of the sevoflurane stated on the vaporizer setting. Moreover, the gas monitor module was out of order, and the heart rate and blood pressure were stable. As a result, we were unaware of the low sevoflurane concentration. The leakage through the hole could not be detected with the commonly used low-pressure system leak checking method. The implication of this case is that unexpected awakening can occur in patients with stable vital signs with an inhalation anesthesia. Therefore, more attention is needed to detect the level of patient awareness.
Anesthesia, General* ; Anesthesia, Inhalation ; Blood Pressure ; Heart Rate ; Humans ; Nebulizers and Vaporizers* ; United Nations ; Vital Signs

PURPOSE: It has been suggested that the pineal hormone melatonin(MEL) protects neurons in vitro from excitotoxicity mediated by kainate-sensitive glutamate receptors and from oxidative stress-induced DNA damage and apoptosis. The present study evaluated the antioxidatives and anti-inflammatory effect of melatonin on kainic acid(KA)-induced neuronal injury in the hippocampus in vivo. METHODS: 30 adult male Sprague-Dawley rats were divided into two equal groups. Control group was treated with KA only and test group was treated with KA and MEL. We injected 10 mg/kg KA intraperitoneal into rats. This results in selective neuronal injury accompanied by intense microglial activation and triggers DNA damage in the hippocampus. We tested the in vivo efficacy of MEL in preventing KA-induced neuronal injury and neuroinflammation in the hippocampus. MEL(2.5 mg/kg) was injected i.p. four times : 20 min before KA, immidiately after, and 1 and 2 h after the KA. Rats were sacrificed 72 h later and their hippocampi were examined for evidence of DNA damage (in situ dUTP-end-labeling, i.e. TUNEL staining), cell viability(H&E staining), microglial (isolectin-B4 histochemistry), astroglial responses(glial fibrillary acidic protein, GFAP immunohistochemistry), and lipid peroxidation(4-hydroxynonenal immunohistochemistry). RESULTS: The cumulative dose of 10 mg/kg MEL attenuates KA-induced neuronal death as well as microglial activation and lessens DNA breaks. CONCLUSION: A possible mechanism of MEL-provided neuroprotection lies in its antioxidant and anti-inflammatory action. Present data suggest that MEL holds potential for the treatment of acute brain pathologies such as epilepsy-associated brain damage, stroke, and brain trauma.
Adult ; Animals ; Apoptosis ; Brain ; Brain Injuries ; DNA Breaks ; DNA Damage ; Epilepsy ; Hippocampus ; Humans ; In Situ Nick-End Labeling ; Kainic Acid ; Male ; Melatonin* ; Neurons* ; Pathology ; Rats ; Rats, Sprague-Dawley ; Receptors, Glutamate ; Stroke

BACKGROUND: A growing body of evidence suggests that neuroinflammation, which is characterized by infiltration of immune cells, activation of mast cells and glial cells, and production of inflammatory mediators in the peripheral and central nervous systems, plays an important role in the induction and maintenance of chronic pain. Palmitoylethanolamide (PEA), which is a type of N-acylethanolamide and a lipid, has an anti-inflammatory effect. Relative to the anti-inflammatory effect, little is known about its analgesic effect in chronic pain. This study aimed to determine whether PEA relieves chronic inflammatory and neuropathic pain. METHODS: Male Sprague-Dawley rats were injured by transection of the left L5 and L6 spinal nerves to induce neuropathic pain or were injected with monoiodoacetic acid into the synovial cavity of knee joints to induce inflammatory pain. To assess the degree of pain, two kinds of stimuli - pressing von Frey filaments and wetting with acetone - were applied to the plantar surface of the rat to measure mechanical and cold sensitivity, respectively. Pain was measured by assessing behavioral responses, including paw withdrawal response threshold and paw withdrawal frequency upon stimulation. RESULTS: Neuropathic pain caused by spinal nerve transection (SNT) decreased the mechanical threshold and increased the frequency of response to acetone application. But, cold allodynia caused by SNT did not decrease the withdrawal frequency. Mechanical hyperalgesia caused by chronic inflammation was significantly reduced by both intraperitoneal and intra-articular injections of PEA. CONCLUSIONS: These outcomes revealed that PEA might be effective in relieving inflammatory and neuropathic pain, especially pain induced by mechanical hyperalgesia, but not cold allodynia.
Acetone ; Animals ; Central Nervous System ; Chronic Pain ; Humans ; Hyperalgesia ; Inflammation ; Injections, Intra-Articular ; Iodoacetic Acid ; Knee Joint ; Male ; Mast Cells ; Neuralgia* ; Neuroglia ; Peas ; Rats* ; Rats, Sprague-Dawley ; Spinal Nerves

Systolic anterior motion of the mitral valve (SAM) is well known in the concentric left ventricular hypertrophy or post mitral valvuloplasty. However, SAM has not been reported in Off-pump coronary artery bypass surgery (OPCAB). Preoperatively, SAM in combination with a left ventricular outflow tract obstruction leads to severe cardiovascular destabilization. Moreover, a diagnosis of SAM is very important because the administration of conventional therapy to hypotension can aggravate SAM. We report a patient with un-identified left ventricular wall hypertrophy or mitral valve regurgitation, who was diagnosed with SAM by TEE during OPCAB. This report describes the diagnostic and therapeutic strategies for the perioperative management of SAM.
Coronary Artery Bypass, Off-Pump* ; Diagnosis ; Humans ; Hypertrophy ; Hypertrophy, Left Ventricular ; Hypotension* ; Mitral Valve Insufficiency ; Mitral Valve*

PURPOSE: The purpose of this study was to investigate the effects of the immunosuppressants FK506 and cyclosporin A (CsA) on the osteogenic differentiation of rat mesenchymal stem cells (MSCs). METHODS: The effect of FK506 and CsA on rat MSCs was assessed in vitro. The MTT assay was used to determine the deleterious effect of immunosuppressants on stem cell proliferation at 1, 3, and 7 days. Alkaline phosphatase (ALP) activity was analyzed on days 3, 7, and 14. Alizarin red S staining was done on day 21 to check mineralization nodule formation. Real-time polymerase chain reaction (RT-PCR) was also performed to detect the expressions of bone tissue-specific genes on days 1 and 7. RESULTS: Cell proliferation was promoted more in the FK506 groups than the control or CsA groups on days 3 and 7. The FK506 groups showed increased ALP activity compared to the other groups during the experimental period. The ALP activity of the CsA groups did not differ from the control group in any of the assessments. Mineralization nodule formation was most prominent in the FK506 groups at 21 days. RT-PCR results of the FK506 groups showed that several bone-related genes-osteopontin, osteonectin, and type I collagen (Col-I)-were expressed more than the control in the beginning, but the intensity of expression decreased over time. Runx2 and Dlx5 gene expression were up-regulated on day 7. The effects of 50 nM CsA on osteonectin and Col-I were similar to those of the FK506 groups, but in the 500 nM CsA group, most of the genes were less expressed compared to the control. CONCLUSIONS: These results suggest that FK506 enhances the osteoblastic differentiation of rat MSCs. Therefore, FK506 might have a beneficial effect on bone regeneration when immunosuppressants are needed in xenogenic or allogenic stem cell transplantation to treat bone defects.
Alkaline Phosphatase ; Animals ; Anthraquinones ; Bone Regeneration ; Cell Differentiation ; Cell Proliferation ; Collagen Type I ; Cyclosporine ; Durapatite ; Gene Expression ; Immunosuppressive Agents ; Mesenchymal Stromal Cells ; Osteoblasts ; Osteonectin ; Rats ; Real-Time Polymerase Chain Reaction ; Stem Cell Transplantation ; Stem Cells ; Tacrolimus

DNA methylation may regulate gene expression by restricting the access of transcription factors. We have previously demonstrated that GATA-1 regulates the transcription of the CCR3 gene by dynamically interacting with both positively and negatively acting GATA elements of high affinity binding in the proximal promoter region including exon 1. Exon 1 has three CpG sites, two of which are positioned at the negatively acting GATA elements. We hypothesized that the methylation of these two CpGs sites might preclude GATA-1 binding to the negatively acting GATA elements and, as a result, increase the availability of GATA-1 to the positively acting GATA element, thereby contributing to an increase in GATA-1-mediated transcription of the gene. To this end, we determined the methylation of the three CpG sites by bisulfate pyrosequencing in peripheral blood eosinophils, cord blood (CB)-derived eosinophils, PBMCs, and cell lines that vary in CCR3 mRNA expression. Our results demonstrated that methylation of CpG sites at the negatively acting GATA elements severely reduced GATA-1 binding and augmented transcription activity in vitro. In agreement, methylation of these CpG sites positively correlated with CCR3 mRNA expression in the primary cells and cell lines examined. Interestingly, methylation patterns of these three CpG sites in CB-derived eosinophils mostly resembled those in peripheral blood eosinophils. These results suggest that methylation of CpG sites at the GATA elements in the regulatory regions fine-tunes CCR3 transcription.
Binding Sites ; Cell Line ; *CpG Islands ; DNA Methylation ; Enhancer Elements, Genetic ; Eosinophils/cytology/*metabolism ; Exons ; Fetal Blood/cytology/metabolism ; GATA1 Transcription Factor/*genetics/metabolism ; Gene Expression Regulation ; Humans ; Promoter Regions, Genetic ; RNA, Messenger/metabolism ; Receptors, CCR3/*genetics/metabolism ; Sequence Analysis, DNA ; *Transcription, Genetic