Octopamine release has been demonstrated from DUMETi in response to high-[K + ] saline and to neuronal stimulation Morton and Evans, 1984 ; . Both forms of octopamine release were shown to be sensitive to the concentration of Ca2 + in the medium bathing the extensor muscle. In addition, the release of octopamine mediated by DUMETi stimulation was shown to be frequency-dependent. Preliminary evidence has been provided for the existence of presynaptic autoreceptors for octopamine on the terminals of DUMETi Morton and Evans, 1984 ; . Evidence has also been provided for the presence of octopaminergic autoreceptors on the cell bodies of cockroach DUM cells from the fifth abdominal ganglion, where they modulate ionic currents and spiking frequency Achenbach et al. 1997 ; . Here, we report on the pharmacological characterization of the presynaptic inhibitory octopamine autoreceptors on the DUMETi terminals and on the specificity of the presynaptic modulation of octopamine release from this preparation. A brief account of some of this work has already been published in abstract form Evans and Howell, 1997 ; . Materials and methods Animals Adult female locusts Schistocerca gregaria ; , 714 days after their final moult, were used in all experiments since the extensor-tibiae muscles of female locusts have been shown to contain higher levels of octopamine than those of males Morton and Evans, 1983 ; . Animals were obtained from crowded laboratory cultures fed on wheat seedlings. Small batches of animals were left undisturbed for 12 days after removal from the main culture before use to avoid any depletion of the octopamine levels in their extensor-tibiae muscles Davenport and Evans, 1984a, b; Evans, 1981 ; . Incubation of muscles Extensor-tibiae muscles from locust hindlegs were removed from the femur and rinsed in saline containing in mmol l-1 ; : 150 NaCl, 6 CaCl2, 4 KHCO3, 4 KH2PO4 and 2 MgCl2 Usherwood and Grundfest, 1965; modified by Evans, 1978 ; . Groups of six muscles were placed in 1 ml saline for 3 min, and the saline was then removed normal saline sample ; . The saline was replaced with 1 ml of high-[K + ] saline high-[K + ] saline sample ; containing various concentrations of K + 0.05, 0.1 or 0.15 mol l-1 K + ; . Ca2 + -free saline was obtained by substitution of Ca2 + with 0.006 mol l-1 Mg2 + , and 0.01 mol l-1 Co2 + saline was obtained by the addition of 0.01 mol l-1 Co2 + to normal saline. In experiments to investigate the effectiveness of drugs on octopamine release, drugs were included in both the normal saline sample and the high-[K + ] saline sample at a concentration of 10-4 mol l-1. In experiments to investigate the time course of octopamine release, 1 min sample periods were used and three exposures of 1 min each to normal saline were followed by three exposures of 1 min each to high-[K + ] saline. Octopamine extraction Octopamine was extracted from the saline samples by passing them through an ion-exchange column Dowex 50W, 100200 mesh size, 2.56 cm0.5 cm internal diameter, H + form ; . The column was washed using 5 ml of water before octopamine was eluted with 5 ml of ammonium hydroxide and lyophilized under vacuum Savant ; . The residue was redissolved in 25 l 0.01 mol l-1 formic acid. The samples were stored frozen until assayed. Radioenzymatic assay for released octopamine The radioenzymatic assay for octopamine used in this study was based on that described by Molinoff et al. 1969 ; as modified by Evans 1978 ; . It resulted in a sensitivity of twice background levels for the assay of 1015 pg 0.1 pmol ; based on a DL-octopamine standard. The assay is based on the amount of radiolabelled N-methyl-octopamine synephrine ; produced from endogenous octopamine and [methyl-3H]S-adenosyl methionine by the enzyme Using an [3H]octopamine standard, the recovery from the Dowex columns was greater than 80 % for all the types of saline used. The authenticity of the radioenzyme assay products was confirmed by thin-layer chromatography as described by Evans 1978 ; , and the presence of drugs in the saline did not interfere with the assay. Chemicals methionine was obtained from Amersham. unlabelled S-adenosyl methionine, DL-octopamine, octopamine agonists and antagonists were obtained from Sigma. Where compounds were not water-soluble, the compound was first dissolved in dimethyl sulphoxide DMSO ; and appropriate controls carried out. The final concentration of DMSO was never greater than 0.1 % [Methyl-3H]S-adenosyl Results K + -stimulated release The K + -stimulated release of endogenous octopamine from the terminals of the DUMETi neurone on the locust extensortibiae muscle was characterized Fig. 1 ; . The amount of octopamine released was dependent upon the concentration of [K + the medium. The release plateaued at a K concentration of 0.1 mol l-1 Fig. 1A ; , and this concentration was used in all subsequent experiments. Fig. 1B shows the time course of the release of endogenous octopamine by 0.1 mol l-1 K + . A low background level of release was found in the first three 1 min sample periods in control saline. In the presence of a high [K + ], the maximum amount of octopamine was released in the first minute of stimulation and the amount released then subsequently declined in the following two sample periods. Thus, in the rest of the present study, the amount of octopamine released during a 3 min period of exposure to 0.1 mol l-1 K + was used to characterize the pharmacology of the response. The K + -stimulated release of octopamine was Ca2 + -sensitive Fig. 1C ; . It was blocked by the removal of external Ca2 + 0Ca2 + ; and by the addition of.
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And the benchmark high-performance liquid chromatography assays29 and liquid chromatography mass spectroscopy assays.30 The chemiluminescence protein-binding assay and the radioimmunoassay are most commonly used to determine patient vitamin D status. Recent reports have raised concerns about the degree of variability between assays and between laboratories, even when using the same assay.29-33 Although reliable and consistent evaluation of serum 25 OH ; D levels remains an issue, reliable laboratories currently exist, and efforts are in progress to improve and standardize assays to enhance accuracy and reproducibility at other laboratories.30, 32, 33 As noted previously, vitamin D plays a central role in calcium and phosphorus homeostasis and skeletal health. Since impaired calcium metabolism due to low serum 25 OH ; D levels triggers secondary hyperparathyroidism, increased bone turnover, and progressive bone loss, 1, 34-38 the optimal range of circulating 25 OH ; D for skeletal health has been proposed as the range that reduces PTH levels to a minimum9, 11, 35 and calcium absorption is maximal.28 Several studies have shown that PTH levels plateau to a minimum steady-state level as serum 25 OH ; D levels approach and rise above approximately 30 ng mL nmol L ; 9, 30, 35-38 Figure 2, left ; . EPIDEMIOLOGY OF VITAMIN D INADEQUACY Vitamin D inadequacy constitutes a largely unrecognized epidemic in many populations worldwide.39-47 It has been reported in healthy children, 7, 8, 13, young adults, 38, 39, 49 especially African Americans, 7, 41, 42, and middle-aged and elderly adults.9-12, 14, 36, 37, Typically, the prevalence of low 25 OH ; D levels 20 ng mL [50 nmol L] ; is approximately 36% in otherwise healthy young adults aged 18 to 29 years, 49 42% in black women aged 15 to 49 years, 50 41% in outpatients aged 49 to 83 years, 11 up to 57% in general medicine inpatients in the United States, 57 and even higher in Europe 28%-100% of healthy and 70%-100% of hospitalized adults ; .40, 55, 58 Vitamin D inadequacy is particularly common among patients with osteoporosis Table 1 ; . A recent systematic review by Gaugris et al39 concluded that the prevalence of inadequate 25 OH ; D levels appears to be high in postmenopausal women and especially those with osteoporosis and a history of fracture. This review, which included 30 studies published between January 1994 and April 2004, examined the prevalence of vitamin D inadequacy reported as serum 25 OH ; D levels below various values. The results of a recent cross-sectional, observational study conducted at 61 sites across North America showed that 52% of postmenopausal women receiving therapy for osteoporosis had 25 OH ; D levels of less than 30 ng mL nmol L ; .30 and tylenol.
For the past fifteen years I have worked as a molecular physiologist researching ion channels in the Department of Physiology at the University of Bristol. Since 2002 I have had 21 papers, 8 of which were either first or last author; all but one paper appeared in journals with impact factors 3. I have just had a first-author paper appear in Molecular Pharmacology Impact Factor 5.48 ; , and in the last year I was a middle-author on papers appearing in Cancer Research Impact Factor 8.65 ; and Biological Psychiatry Impact Factor 6.04 ; . I single-handedly set up molecular biology in my department. Within my department: I created the first functioning molecular laboratory, published the first molecular paper of any kind, cloned the first full length gene transcript, created the first inducible cell line, and published the first paper on heterologous expression. I have written five successfully funded grants since 1998, with total allocations of 550, 047.
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Pressure. This would create tension on the retina, and in a susceptible patient could cause a choroidal hemorrhage. I discussed this scenario with one of my surgeons, who stated that this is not only a definite possibility, but that he has heard of that happening without involving a block. Factors that would make such a patient susceptible to a choroidal hemorrhage include hypertension, advanced age, macular degeneration, anticoagulation, and taking at least one cardiovascular medication. My references for this are RETINA, April, 2003, Yang, and Br. J. Ophthalmology, April, 2004, Ling ; . This patient had at least one of these factors. From her description, the block technique was fine, the patient's anatomy was normal, and this was just a most unfortunate cascade of events. Terry Gabrielson, MD, Rancho Mirage, CA Answer #3 We read this case with great interest. There are a number of relevant ophthalmic anesthesia issues. These include regional anesthesia for traumatized eyes, needle-globe penetration, choroidal hemorrhage, and perioperative patient movement. Over the past few years we have used regional anesthesia selectively for patients whose eyes have sustained open-globe injuries. Our experience has been published Ophth Surg Lasers & Imaging 36 #2: 122 128, 2005 and AJO 134 #5: 707 711, 2002 ; . While this patient presumably did not have a dehiscence prior to the block, a failed graft is reason enough to be extra cautious during the block. During injection, the lesion is readily visualized for evidence of elevated intraocular pressure. The precise technique used for anesthesia intraconal vs. extraconal ; is indeterminable from the vignette. Nonetheless, it is our impression that ocular force volume curves seem to be independent of type of block, but more dependent upon speed of injection and intrinsic patient factors orbital volume, globe size, and tightness laxity of tissues.
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Exchange Risk Elan is a multinational business operating in many countries. The US dollar is the primary currency in which Elan conducts its business. The US dollar is used for planning and budgetary purposes and as the currency for financial reporting. Elan has revenues, costs, assets and liabilities denominated in currencies other than US dollars. The Group manages its non-US dollar foreign exchange risk through derivative financial instruments. The US dollar is the base currency against which all identified transactional foreign exchange exposures are managed and hedged. The principal risks to which Elan is exposed are movements in the exchange rates of the US dollar against the Euro, Sterling, Swiss Franc and Japanese Yen. The main exposures are net costs in Euro arising from a manufacturing and research presence in Ireland and the sourcing of raw materials in European markets. During 2001, average exchange rates were EUR1.1187 US$1. Elan sells US dollars to buy Euro for costs incurred in Euro. The expected strengthening of the Euro against the US dollar will result in a higher reported cost related to Elan's Euro cost base in 2002 compared to 2001. However, Elan does not expect this to be material. All derivative contracts entered into are in liquid markets with credit approved counterparties. For additional information regarding foreign exchange risk, please refer to Note 21 to the Consolidated Financial Statements. Interest Rate Risk Elan's liquid funds are invested primarily in US dollars except for the working capital balances of subsidiaries operating outside of the United States. Interest rate risk is mainly confined to the variability of returns on investment funds as the majority of Elan's debt is fixed rate. The Group's exposure to interest rate risk is actively monitored and managed with an average duration of less than three months. By calculating an overall exposure to interest rate risk rather than a series of individual instrument cash flow exposures, the Company can more readily monitor and hedge these risks. Duration analysis recognises the time value of money and in particular, prevailing interest rates by discounting future cash flows. For additional information regarding interest rate risk, please refer to Note 21 to the Consolidated Financial Statements. Credit Risk Elan's treasury function transacts business with counterparties that are considered to be low investment risk. Credit limits are established commensurate with the credit rating of the financial institution that business is being transacted with. Elan does not believe that it has a significant exposure to any one financial counterparty. Elan does not currently transact significant business in countries that are subject to political and economic uncertainty. As a result, Elan is not materially exposed to any sovereign risk or payment difficulties. Liquidity Risk For additional information regarding liquidity risk and for sensitivity analysis information, please refer to Note 21 to the Consolidated Financial Statements. Equity Price Risk US GAAP ; Elan is exposed to equity price risks primarily on its available for sale securities which consist of equity investments in quoted companies. At 31 December 2001, available for sale securities had a fair value of $305.3 million and had a cost of $282.1 million. These investments are primarily in emerging pharmaceutical and biotechnology companies. A 10% adverse change in equity prices would result in an approximate $30.5 million decrease in the fair value of Elan's available for sale equity securities.
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