OBJECTIVE: The brain is dependent on glucose as an energy source. Intricate homeostatic mechanisms have been implicated in maintaining the blood glucose concentration in the brain. The aim of this study is to find the way to identify the metabolic proteins regulating the glucose in rat hypothalamus. METHODS: In this study, we analysed the secretome from rat hypothalamus in vivo. We introduced 500 nM of insulin into the rat hypothalamus. The chromatographic patterns of the secretome were identified, after which Mass Spectrometry-Mass Spectrometry (MS-MS) analysis was performed. RESULTS: In Liquid Chromatography-Mass Spectrometry (LC-MS) analysis, 60 proteins were identified in the secretome. Among them, 8 novel proteins were unveiled and were associated with the energy metabolism of insulin signaling in mitochondria of rat hypothalamic neuron. Nineteen other proteins have unknown functions. These ligands were confirmed to be secreting from the rat hypothalmus on insulin signaling by western blotting. CONCLUSION: The hypothalamus is the master endocrine gland responsible for the regulation of various physiological and metabolic processes. Proteomics using LC-MS analysis offer a efficient means for generating a comprehensive analysis of hypothalamic protein expression by insulin signaling.
Animals ; Blood Glucose ; Blotting, Western ; Brain ; Chromatography, Liquid ; Endocrine Glands ; Energy Metabolism ; Glucose ; Hypothalamus ; Insulin* ; Ligands ; Mass Spectrometry ; Metabolism ; Mitochondria ; Neurons ; Proteomics ; Rats* ; Spectrum Analysis* ; Tandem Mass Spectrometry

The aim of this study is to identify hRad21-binding sites in human chromosome, the core component of cohesin complex that held sister chromatids together. After chromatin immunoprecipitation with an hRad21 antibody, it was cloned the recovered DNA and sequenced 30 independent clones. Among them, 20 clones (67%) contained repetitive elements including short interspersed transposable elements (SINE or Alu elements), long terminal repeat (LTR) and long interspersed transposable elements (LINE), fourteen of these twenty (70%) repeats clones had Alu elements, which could be categorized as the old and the young Alu Subfamily, eleven of the fourteen (73%) Alu elements belonged to the old Alu Subfamily, and only three Alu elements were categorized as young Alu subfamily. There is no CpG island within these selected clones. Association of hRad21 with Alu was confirmed by chromatin immunoprecipitation-PCR using conserved Alu primers. The primers were designed in the flanking region of Alu, and the specific Alu element was shown in the selected clone. From these experiments, it was demonstrated that hRad21 could bind to SINE, LTRs, and LINE as well as Alu.
Alu Elements ; Chromatids ; Chromatin ; Chromatin Immunoprecipitation ; Chromosomes, Human* ; Clone Cells ; CpG Islands ; DNA ; DNA Transposable Elements ; Humans ; Humans* ; Siblings ; Terminal Repeat Sequences

Serotonin (5-HT), the endogenous nonselective 5-HT receptor agonist, activates the inositol -1,4,5- triphosphate / calcium (InsP3/Ca2+) signaling pathway and exerts both stimulatory and inhibitory actions on cAMP production and neuromodulin secretion in rat hypothalamic neurons. Specific mRNA transcripts for 5-HT1A, 5-HT2C and 5-HT4 were identified in rat hypothalamic neurons. These experiments were supported by combined techniques such as cAMP and a Ca2+ assays in order to elucidate the associated receptors and signaling pathways. The cAMP production and neuromodulin release were profoundly inhibited during the activation of the Gi-coupled 5-HT1A receptor. Treatment with a selective agonist to activate the Gq-coupled 5-HT2C receptor stimulated InsP3 production and caused Ca2+ release from the sarcoplasmic reticulum. Selective activation of the Gs-coupled 5-HT4 receptor also stimulated cAMP production, and caused an increase in neuromodulin secretion. These findings demonstrate the ability of 5-HT receptor subtypes expressed in neurons to induce neuromodulin production. This leads to the activation of single or multiple G-proteins which regulate the InsP3/Ca2+/PLC-gamma and adenyl cyclase / cAMP signaling pathways.
Animals ; Calcium ; GAP-43 Protein* ; GTP-Binding Proteins ; Inositol ; Neurons* ; Rats* ; Receptor, Serotonin, 5-HT1A ; Receptor, Serotonin, 5-HT2C ; Receptors, Serotonin, 5-HT4 ; RNA, Messenger ; Sarcoplasmic Reticulum ; Serotonin* ; Adenylyl Cyclases

Infection stones are more likely to form after urinary diversion as the result of urinary stasis. To prevent urinary stasis due to encrusted pyelitis in a transplanted kidney, we describe an alternative a surgical treatment: ileo-pelvic anastomosis. In our patient with a transplanted kidney, the ileal conduit had previously been anastomosed end-to-side owing to renal tuberculosis with an atrophied bladder; the transplanted ureter was anastomosed to the ileum in the left lower abdomen with an ileal conduit on the opposite side. Routine check-up revealed hydronephrosis with infected pyelitis and ureteritis in the transplanted kidney. We performed ileo-pelvic end-to-end anastomosis to prevent urinary stasis by lengthening the ileal conduit and performed augmentation cystoplasty to support the atrophied bladder following tuberculosis. We suggest that this approach may be useful in similar cases.
Abdomen ; Humans ; Hydronephrosis ; Ileum ; Kidney ; Kidney Transplantation ; Pyelitis ; Transplants ; Tuberculosis ; Tuberculosis, Renal ; Ureter ; Urinary Bladder ; Urinary Diversion

Here we determined which radiation-responsive genes were altered in radioresistant CEM/IR and FM3A/IR variants, which showed higher resistance to irradiation than parental human leukemia CEM and mouse mammary carcinoma FM3A cells, respectively and studied if radioresistance observed after radiotherapy could be restored by inhibition of protein kinase A. The expressions of DNA-PKcs, Ku70/80, Rad51 and Rad54 genes that related to DNA damage repair, and Bcl-2 and NF-kappaB genes that related to antiapoptosis, were up-regulated, but the expression of proapototic Bax gene was down-regulated in the radioresistant cells as compared to each parental counterpart. We also revealed that the combined treatment of radiation and the inhibitor of protein kinase A (PKA) to these radioresistant cells resulted in synergistic inhibition of DNA-PK, Rad51 and Bcl-2 expressions of the cells, and consequently restored radiosensitivity of the cells. Our results propose that combined treatment with radiotherapy and PKA inhibitor can be a novel therapeutic strategy to radioresistant cancers.
Animals ; Apoptosis/drug effects ; Cell Line, Tumor ; Cyclic AMP-Dependent Protein Kinases/*antagonists & inhibitors/metabolism ; DNA Damage/drug effects ; DNA Repair/drug effects ; Gamma Rays ; Gene Expression Regulation, Neoplastic/radiation effects ; Genes, bcl-2 ; Humans ; Mice ; Neoplasm Proteins/genetics/metabolism ; Neoplasms/enzymology/*genetics ; Radiation Tolerance/*genetics ; Research Support, Non-U.S. Gov't