ABSTRACT
A new method of stabilizing the nasal endotrascheal tube was described. The tube was secured to the anterior portion of the nasal septum with braided silk thread, which was tightened over the rubber tube to keep air route of the cuff open. There found no complications such as unplanned extubation, necrosis and infections of the nasal septum.
Subject(s)
Intubation, Intratracheal/instrumentation , Intubation, Intratracheal/methods , Nasal Septum/surgery , Sutures , Adult , Aged , Aged, 80 and over , Equipment Design , Female , Humans , Insect Proteins , Male , Middle Aged , Rubber , SilkABSTRACT
PURPOSE: Our purpose was to determine the association of calmodulin-dependent protein kinase II (CaMKII) with oocyte activation and to explore the network of protein kinases during mammalian fertilization. METHODS: Mouse M-II oocytes were collected after superovulation induced by PMSG-hCG injection. The oocytes were inseminated or artificially activated by Ca ionophore (A23187) or 12-O-tetradecanoyl phorbol 13-acetate (TPA). The effects of KN-62, a specific and selective inhibitor of calmodulin-dependent protein kinase II, on second polar body emission (2PBE), pronuclear formation (PF), and cortical granule exocytosis (CGE) during fertilization or after artificial oocyte activation were investigated. RESULTS: KN-62 inhibited 2PBE and PF after sperm or Ca ionophore inducing activation. Additionally, PF was inhibited by KN-62 after TPA activation, whereas KN-62 did not inhibit CGE in any case. KN-04, an inactive form of KN-62, did not inhibit significantly 2PBE, CGE, or PF. When oocytes were exposed to KN-62 after Ca ionophore or TPA activation, no inhibitory effects on 2PBE or PF were observed. CONCLUSIONS: The CaMKII activation that occurs after fertilization or artificial activation of mouse oocytes is presumably secondary to increases in the intracellular free calcium concentration. As determined by the use of inhibitor, CaMKII activity is associated with 2PBE and PF but not with CGE.
Subject(s)
1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine/analogs & derivatives , Calcium-Calmodulin-Dependent Protein Kinases/antagonists & inhibitors , Calcium-Calmodulin-Dependent Protein Kinases/physiology , Enzyme Inhibitors/pharmacology , Oocytes/physiology , 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine/analysis , 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine/pharmacology , Animals , Calcimycin/pharmacology , Female , Fertilization in Vitro , Fluorescent Dyes , Ionophores/pharmacology , Male , Mice , Mice, Inbred ICR , Oocytes/drug effects , Protein Kinase InhibitorsABSTRACT
Activation of oocytes is caused by osmotic pressure change in some species. However, cryopreservation of oocytes occurs in the presence of osmotic pressure change induced by cryoprotectants. We investigated the effect of 5-(N,N,-dimethyl)-amiloride (NNDMA), a selective inhibitor of Na+/H+ exchange, on the cryopreservation and osmotic activation of mouse oocytes. The percentage (23.2%) of degenerate oocytes after cryopreservation in the presence of NNDMA was found to be lower than that (39.5%) of untreated oocytes. After thawing, the percentage (23.6%) of oocytes which could be fertilized following cryopreservation in the presence of NNDMA was significantly higher than that of untreated (18.0%) oocytes. These results suggest that amiloride increased the survival rate after thawing following cryopreservation. To investigate the effect of NNDMA on oocyte activation caused by the cryoprotectant, dimethyl sulphoxide (DMSO) was used to induce osmotic pressure change. NNDMA was found to inhibit cortical granule exocytosis, the second polar body emission and pronuclear formation which occurs upon activation due to osmotic pressure change. It also inhibited the increase in phosphorylation of many proteins including 33 and 45 kDa proteins, which occurs, during fertilization and chemical oocyte activation. In contrast, protein phosphorylation was not inhibited by W7, a calmodulin inhibitor. The actions of these inhibitors suggest that oocyte activation induced by osmotic pressure change involves a pathway mediated by Na+/H+ exchange which may be distinct from the Ca-calmodulin pathway. Amiloride may be a useful drug for increasing the rate of survival of cryopreserved oocytes.
