IWALT 2000 Panel – 2 Telemedicine and Tele- Education Remote Tele-Education as an Alternat
《当代医学英语综合教程I——医学探索(第二版)》

当代医学英语综合教程I——医学探索(第二版)引言当代医学英语综合教程I——医学探索(第二版)是针对医学专业学生开发的一本教材。
本教程旨在提供全面、系统的医学英语学习资源,帮助学生提高医学英语的听、说、读、写能力。
本文档将对《当代医学英语综合教程I——医学探索(第二版)》进行介绍,并展示部分教材的内容。
教材概述作者本教材的主要作者是医学教育专家李华和英语教育专家John Smith。
李华拥有丰富的医学教学经验,曾在多所医学院授课。
John Smith则在英语教育领域有着卓越的贡献,他参与过多个医学英语教材的撰写。
内容该教材共分为十个单元,涵盖了医学英语的各个方面。
每个单元包括了课文、词汇、语法和练习等内容,帮助学生从多个角度进行学习。
同时,教材还提供了翻译和解析,帮助学生快速理解和运用所学知识。
下面将展示教材中的一个单元作为示例。
单元示例:Unit 4 Medical Conditions课文Text 1: Cardiovascular DiseasesCardiovascular diseases are a class of diseases that involve the heart or blood vessels. They include coronary artery disease, heart failure, valvular heart disease, and more. Cardiovascular diseases are the leading cause of death globally.Text 2: Respiratory DisordersRespiratory disorders affect the organs and tissues involved in breathing. Common respiratory disorders include asthma, chronic obstructive pulmonary disease (COPD), pneumonia, and lung cancer. Proper diagnosis and treatment are crucial for managing respiratory disorders.词汇•cardiovascular diseases(心血管疾病)•coronary artery disease(冠状动脉疾病)•heart failure(心力衰竭)•valvular heart disease(心脏瓣膜疾病)•respiratory disorders(呼吸系统疾病)•asthma(哮喘)•chronic obstructive pulmonary disease (COPD)(慢性阻塞性肺疾病)•pneumonia(肺炎)•lung cancer(肺癌)语法在本单元的语法部分,学生将学习关于医学条件描述的语法结构。
雅培Architect i2000化学发光分析仪检测乙型肝炎病毒标志物的应用

雅培Architect i2000化学发光分析仪检测乙型肝炎病毒标志物的应用花艳艳;李梦华;丁楠【摘要】目的:研究探讨雅培Architect i2000化学发光分析仪检测乙肝病毒标志物,并且与传统ELISA法检测效果进步对比分析.方法:对2587例患者的血样血清分别采用ELISA法和化学发光法检测五种乙肝病毒标志物(HBsAg、HBsAb、HBeAg、HBeAb、HBcAg)水平.结果:化学发光分析法检测五种乙肝病毒标志物符合率分别为:92.2%、95.0%、91.1%、96.1%、94.4%,其中乙肝病毒阳性检出率为6.92%;而ELISA法检测五种乙肝病毒标志物符合率分别为:81.0%、84.9%、83.2%、86.6%、83.8%,阳性检出率则为:5.72%.结论:相比于传统的ELISA法,化学发光法具有更高的精确性、稳定性、重复性以及较低的环境因素影响等,是临床上检测乙肝病毒的重要方法之一,其中使用雅培Architect i2000化学发光分析仪使得检测数据更准确、可靠.【期刊名称】《中国医疗器械信息》【年(卷),期】2018(024)012【总页数】2页(P75-76)【关键词】乙型肝炎标志物;化学发光法;ELISA法【作者】花艳艳;李梦华;丁楠【作者单位】大连市第六人民医院检验科,辽宁大连 116000;大连市第六人民医院检验科,辽宁大连 116000;大连市第六人民医院检验科,辽宁大连 116000【正文语种】中文【中图分类】R512.62乙型肝炎病毒(Hepatitis B Virus,HBV),又简称乙肝病毒,是一种DNA病毒,属于嗜肝性DNA病毒科[1]。
据统计,到目前为止,全世界感染慢性乙肝病毒患者约为4亿左右,每年死于乙肝病毒及其并发症的患者约有100万[2,3]。
临床上乙肝病毒检测标志物主要有:乙肝病毒表面抗原HBsAg,乙肝病毒表面抗原HBsAb,乙肝病毒e抗原HBeAg,乙肝病毒e抗原HBeAb以及乙肝病毒核心抗原HBcAg等[4-6]。
i2000常规操作简介

i2000常规操作1.开机①打开SCC,等待其转到Snapshot画面②打开PM电源(同时接通SH电源),待snapshot屏幕上PM与SH的状态转为stop③选择PM与SH图标,选择F5 Startup,等待两者的状态均转为ready,即可开始每日工作2.每日工作①标准曲线申请:Orders→Calibration order→选定项目,设定位置,输入标准品的批号和效期→F2 Add order②病人样本申请:Orders→Patient order→选定项目,设定位置→F3 Addorder。
如为批试样本,选择Batch,设定起始样本号及位置即可③质控样品的申请:Orders→Control order→选定项目,设定位置,选择批号和质控水平→F2 Add order3.检查供应中心①Supplies→Supply status→检查各部分是否充足,是否需要更换,如需要,选择F2→ Update supplies→更新完成后→Done②放置试剂:新试剂第一次上机时,要先充分混匀,换上septum,按照不同颜色放在试剂转盘的相应位置上即可,放置完成后,不需要每天取出,可上机保存4.运行①将样品、试剂按照需要放置完毕后,即可开始运行,选择snapshot屏幕上PM图标→F8 Run→PM开始初始化,完成后转为Running状态→待PM达Running状态后,再选择SH图标→ F8 Run →SH开始初始化,完成后转为Running状态→仪器开始测定②全部测定完成后,PM保持Running状态,SH转为Load queue paused,可随时选择继续运行5.浏览和处理结果①在运行过程中,选择Orders→order status可浏览当前工作状态,查看某项目是否完成②在运行过程中,选择Results→Result review可浏览当前结果,可选择Release或Print或Rerun或Delete6.查看标准曲线Snapshot→QC.Cal→Calibration status→选定某项目→F5 Details查看详细资料7.不良结果处理Snapshot→Exception→选定某测试项目→点击屏幕右下角的“?”图标,可帮助查找相关资料。
Excessive thyroxine enhances susceptibility to apoptosis and decreases

Molecular and Cellular Endocrinology 320 (2010) 67–75Contents lists available at ScienceDirectMolecular and CellularEndocrinologyj o u r n a l h o m e p a g e :w w w.e l s e v i e r.c o m /l o c a t e /m ceExcessive thyroxine enhances susceptibility to apoptosis and decreases contractility of cardiomyocytesYun-Ying Wang,Bo Jiao,Wang-Gang Guo,Hong-Lei Che,Zhi-Bin Yu ∗Key Laboratory of Aerospace Medicine,Ministry of Education,Department of Aerospace Physiology,Fourth Military Medical University,169#Changlexi Road,Xi’an 710032,Chinaa r t i c l e i n f o Article history:Received 3September 2009Received in revised form 12November 2009Accepted 22January 2010Keywords:HyperthyroidCardiac hypertrophy Heart failure Contractility Apoptosisa b s t r a c tExcessive thyroid hormone induces cardiac hypertrophy and promotes heart failure in patients with hyperthyroidism,but the mechanism remains elusive.Rats were orally administered with levothyroxine (100g/kg,T 4)for 4weeks to induce hyperthyroidism.The calculated stroke volume decreased and the shortening amplitude–frequency relationship in unloaded contraction of isolated cardiomyocytes was negative in T 4-treated rats.Apoptotic rates increased and DNA laddering was also detectable in T 4-treated rat hearts.By contrast,in primary cultured cardiomyocytes,T 3induced dose-dependent hypertrophy but did not affect the apoptotic rate.Angiotensin II further increased the apoptotic rate of T 3-induced hyper-trophied cardiomyocytes.The apoptotic rate was dependent on the extent of cardiomyocyte hypertrophy.These results suggest that cardiac contractility is enhanced during the early stage of hyperthyroidism,but decreased during the late stage of hyperthyroidism.The hypertrophied cardiomyocytes were also suscep-tible to apoptotic stimulation by angiotensin II.Depressed cardiac contractility and enhanced apoptosis may lead to heart failure in hypertrophied myocardium.© 2010 Elsevier Ireland Ltd. All rights reserved.1.IntroductionThyroid hormone (TH)is a key hormone responsible for main-taining cardiac function in vertebrates (Klein and Danzi,2007).TH induces physiological hypertrophy during the fetal-to-adult switch (Bates et al.,1999).Moderate increases in TH levels enhance the expression of sarcoplasmic reticulum Ca 2+-ATPase (Moolman,2002)and promotes the -myosin heavy chain (MHC)-to-␣-MHC switch (Everett et al.,1984),for example.These changes have bene-ficial effects on cardiac performance.Thus,TH treatment was found to improve cardiac function in the post-infarction model (Mahaffey et al.,1995)and converts from pathological to physiological hyper-trophy in pressure-overload animal models (Chang et al.,1997).However,long-term excessive TH levels also results in heart failure (Dahl et al.,2008).The mechanisms of hyperthyroidism-induced heart failure remain uncertain.Hyperthyroidism increases the resting heart rate.Thus,hyperthyroid-induced tachycardia may be responsible for heart failure in hyperthyroidism (Dahl et al.,2008).Excessive TH also induces the development of multifocal areas of fibrosis in the rat heart (Gomberg-Maitland and Frishman,1998).These focal areas of fibrosis may indicate repair after apoptosis or necrosis of cardiomyocytes in the ventricular myocardium.Hyperthyroidism can also induce apoptosis in liver cells (Upadhyay et al.,2004)∗Corresponding author.Tel.:+862984774807;fax:+862983248036.E-mail address:yuzhib@ (Z.-B.Yu).and pancreatic cells (Jorns et al.,2002).However,there is little information about hyperthyroidism-induced apoptosis of car-diomyocytes.Long-term excessive TH enhances the sensitivity of the myocardium to catecholamine and increases the activity of the renin–angiotensin system in the myocardium (Hu et al.,2003).Chronically elevated catecholamine and/or angiotensin II (Ang II)levels in the myocardium reduce cardiac contractility and pro-mote apoptosis of cardiomyocytes in humans and animals (Barry et al.,2008).Thus,hyperthyroidism may indirectly induce apop-tosis or necrosis of cardiomyocytes.However,until now,it was unclear whether hyperthyroid-treated heart or cardiomyocytes show altered cardiac contractility or increased apoptotic rates.The aim of this study was to observe the changes in cardiac contractility and the apoptotic rate in rat hearts after treatment with excessive thyroxine levels.We also assessed the cardiac contractility of isolated working hearts and single cardiomyocyte preparations obtained from 2-and 4-week hyperthyroid-treated rats.Cardiomyocyte apoptosis was detected in the hyperthyroid-treated heart and cultured cardiomyocytes.The contribution of depressed cardiac contractile function and cardiomyocyte apop-tosis to heart failure was analyzed in hyperthyroid-treated rats.2.Materials and methods 2.1.Animal modelHealthy male Sprague-Dawley rats,weighing 180–200g,were housed in Plexiglas cages on a 12:12-h light/dark cycle and received chow and water ad libitum before and during all experiments.Sodium levothyroxine (T 4)was purchased from Merck (Darmstadt,Germany).Rats were randomly divided into0303-7207/$–see front matter © 2010 Elsevier Ireland Ltd. All rights reserved.doi:10.1016/j.mce.2010.01.03168Y.-Y.Wang et al./Molecular and Cellular Endocrinology320 (2010) 67–752-and4-week synchronous control groups,and2-and4-week T4-treated groups. Hyperthyroidism was simulated by daily oral administration of T4(100g/kg body weight)for2or4weeks.Control rats received daily oral administration of saline. Blood pressure was assessed by a tail-cuff in conscious rats before each experiment. The animal procedures were approved by the animal care and use committee at the Fourth Military Medical University in China.2.2.Light microscopy and immunohistochemical detection of apoptotic cardiomyocytesThe heart was infused retrogradely,via aorta cannulation,with20ml of phosphate-buffered saline(PBS),followed by40ml of4%paraformaldehyde in PBS for approximately10min.The heart was thenfixed overnight.After paraffin embed-ding,slices werefixed to glass slides and stained by hematoxylin and eosin.Thefixed longitudinal ventricular slices were embedded in paraffin and10-m thick sections were deparaffinized by washing in xylene and a descending ethanol series.The sections were then incubated with20g/ml proteinase K for 5min at room temperature.The terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL)assay using dUTP-FITC(Apo-BrdU In Situ DNA Fragmentation Assay Kit,BioVision Inc.,USA)was carried out according to the man-ufacturer’s protocols.To specifically identify apoptotic cardiomyocytes,sections were concomitantly stained with a primary antibody against cardiac myosin heavy chain(1:50,Cambridge,UK),followed by incubation with a tetramethyl-rhodamine-labeled secondary antibody(1:400;Molecular Probes)for1h at room temperature. Five hundred myocyte nuclei were examined infive differentfields.The percentage of TUNEL-positive myocyte nuclei was calculated as%apoptosis percentage.2.3.Agarose gel electrophoresis of genomic DNADNA was extracted as previously described(Upadhyay et al.,2004).Approx-imately100mg of heart tissue was homogenized in an equal volume of homogenization buffer(100mM NaCl,5mM ethylenediaminetetraacetic acid (EDTA),100mM Tris–HCl,pH8.0),and centrifuged.The supernatant was trans-ferred to digestion buffer(homogenization buffer plus0.5%sodium dodecyl sulfate (SDS),0.25mg/ml proteinase K)and incubated for2h at37◦C.RNA was removed by RNase treatment(0.25mg/ml RNase,DNase free).DNA was extracted with a phenol/chloroform/isoamyl alcohol(25:24:1)mixture and precipitated overnight at−20◦C with ethanol/sodium acetate.The precipitated DNA was washed with 75%ethanol;vacuum dried and dissolved in TE buffer(100mM Tris–HCl,5mM EDTA,pH8.0)for at least1h at65◦C.The DNA concentration was measured spec-trophotometrically at OD260.A10-g sample of DNA was loaded onto a2.0%agarose gel containing ethidium bromide and electrophoresed at90V.The gels were pho-tographed under UV light.2.4.Preparation of isolated working heartsIsolated working hearts were studied by a technique described elsewhere(Araki et al.,2005).Briefly,rats were injected intraperitoneally with1000IU of heparin for20min,and then anesthetized with sodium pentobarbital(50mg/kg,i.p.).The heart was rapidly excised.The aorta was mounted onto a cannula(internal diame-ter:2.0mm).The heart was immediately perfused in a non-circulating pattern on a Langendorff apparatus(Radnoti,USA)maintaining a pressure of60mmHg with oxy-genated(95%O2/5%CO2,at pH7.40)Krebs–Henseleit solution composed of(in mM) 120.0NaCl,4.7KCl,2.50CaCl2,1.2MgSO4,20.0NaHCO3,1.2NaH2PO4,0.25EDTA and10.0glucose.Connective and lung tissue was carefully removed.The left atrium was cannulated via the pulmonary vein.All pulmonary veins were ligated.When a tight seal without leaks had been established,the heart was switched to work-ing mode after a15-min equilibration period by converting the supply of perfusate from the aorta to left atrial cannula,with afilling(preload)pressure of10mmHg.The preload was varied at5,10,and15mmHg.The working heart ejected the perfusate via the aortic valve into the aortic cannula.The hydrostatic pressure in the aortic cannula was maintained at60mmHg(after-load)throughout the working phase for all rat hearts.Aortic pressure was monitored by a side arm of the aortic cannula with a pressure transducer(MLT844,MEMSCAP AS,Norway).Aorticflow was obtained by collecting the aortic chamber overflow in a graduated cylinder during working conditions.Aortic pressure was recorded by PowerLab-4/25(ADInstruments Inc., Sydney,Australia)and the heart rate was calculated from the left ventricular pres-sure waves.Coronaryflow was assessed by collecting the effluent draining from the apex of the heart at5-min intervals.Cardiac output was calculated as the sum of coronary and aorticflow(i.e.,cardiac output=coronaryflow+aorticflow).Isolated hearts were excluded from the study if initial coronaryflow,aorticflow and heart rate were less than5ml/min,15ml/min,and200beats/min,respectively, or if arrhythmias occurred during the initial perfusion period(30min).Based on these criteria,no hearts were excluded from the present experiment.2.5.Isolation of rat ventricular cardiomyocytesCardiomyocytes were isolated from rat hearts using a previously described tech-nique(Nagata et al.,1998).The heart was rapidly excised and immediately immersed in Ca2+-free Joklik solution(Sigma)containing10mM HEPES and10mM NaHCO3.The heart was perfused in the Langendorff mode with non-circulating Joklik solution at a constantflow of8ml/min for5min and then perfused with the circulating diges-tion Joklik solution containing0.08%collagenase I(Sigma)plus0.1%BSA(Sigma)for 30min.Finally,the digestion Joklik solution was washed out with Joklik solution for 5min.The ventricular myocardium was cut into small pieces and gently agitated, allowing the cardiomyocytes to be dispersed into Joklik solution.The cardiomy-ocytes werefiltered through mesh screens,centrifuged for3min at400×g,and resuspended in fresh Joklik solution containing1%BSA at room temperature.The cell suspension was settled and bubbled slowly with100%oxygen for30min at room temperature.CaCl2was added at5-min intervals to increase the Ca2+concentration to1.25mM.The cells were analyzed within6h after isolation.2.6.Unloaded contractile function of cardiomyocytesThe cardiomyocytes were transferred to a0.3-ml myocyte chamber,which was mounted on the stage of an inverted microscope(OlympusX71,Japan)and settled to the bottom of chamber for10min.Cardiomyocytes were perfused continuously at37◦C with oxygenated Tyrode buffer containing(in mM)132.0NaCl,4.8KCl, 1.2MgCl2,10.0HEPES,5.0sodium pyruvic,1.8CaCl2,pH7.2,at aflow rate of 0.2ml/min.Two platinum electrodes connected to a stimulator(ADInstruments Inc.,Sydney,Australia)were placed on two sides of an identified healthy-appearing, rod-shaped cardiomyocyte with clearly visible striations.The cardiomyocyte was field-stimulated at frequencies of1.0,2.0or4.0Hz(5-ms pulse duration;15V/cm).The signal from a CCD camera was fed to a video-edge detector enabling on-line cardiomyocyte data acquisition using specialist software(PTI,Lawrenceville,USA). Data were acquired at the sample rate of240Hz,stored and analyzed by the PTI soft-ware.The length and width of the cardiomyocytes were quantified by comparison with a calibrated micrometer on the microscope stage.2.7.Primary culture of neonatal rat cardiomyocytesNeonatal rat cardiomyocytes were prepared from the ventricular tissue of1-to 3-day-old Sprague-Dawley rats by a method described elsewhere(Fujio et al.,2000). Ventricles from six rats were aseptically removed and minced into approximately 1-mm3chunks in serum-free Hank’s balanced salt solution.The cardiomyocytes were disaggregated by repeated digestion with0.1%collagenase I dissolved in HEPES buffer(116mM NaCl,0.8mM MgSO4,5.4mM KCl,1.0mM NaH2PO4,5.0mM glucose, 20mM HEPES,pH7.40)at37◦C under continuous mixing with a magnetic stirrer at150rpm.Thefirst tissue digest consisting mainly of cell debris and mesenchy-mal cells was discarded.The following2–5supernatants obtained after each5-min digestion period were collected into single sterile glass centrifuge tube containing warm Dulbecco’s modified Eagle’s medium(DMEM)supplemented with10%new-born calf serum.Cells were gently sedimented at500×g for5min.The cell pellet was resuspended in DMEM supplemented with10%fetal calf serum(FCS),0.1mM 5-bromodeoxyuridine(Sigma),and antibiotics(100U/ml penicillin and100mg/ml streptomycin).To enrich the cardiomyocytes,20ml of this suspension was plated in a10-cm diameter plastic Petri dish,and incubated for90min in5%CO2at37◦C. The cardiomyocyte-enriched suspension was carefully removed and seeded into a six-well plate.Thefinal cell density was adjusted to3.0×105cells per well.The cells were incubated at37◦C in5%CO2incubator overnight and over90%of cells in culture were observed to beat.After48-h incubation,the FCS-supplemented growth medium was replaced by serum-free,hormone-supplemented DMEM medium containing0.1mM5 -bromodeoxyuridine,10g/ml insulin,6.7ng/ml transferrin, 5.5g/ml sodium selenite(Invitrogen),and0.1%bovine serum albumin.After24-h incubation,the synchronous control cells were cultured for a further48h with-out any treatment before morphological and biochemical analysis.Cardiomyocyte hypertrophy was induced by3,3 ,5-triiodo-l-thyronine(T3)for48h.Hypertrophied cells were then cultured with1nM of Ang II for24h.2.8.Quantification of cardiomyocyte surface area and apoptotic assayQuantification of cell size was performed on eosin-stained monolayer-attached cardiomyocytes.The cell surface area was measured by NIH ImageJ software(avail-able at /ij/download.html).Plates were washed in PBS andfixed in4%paraformaldehyde for25min at room temperature.Cells were then permeabilized by0.2%Triton X-100in PBS for 5min at room temperature.After washing with PBS,cells were incubated with the reaction mixture of the terminal TUNEL assay kit(DeadEnd Colorimetric TUNEL System,Promega)at37◦C in a humidified chamber,following the manufacturer’s instructions.As a positive control,cells were treated with DNase I(0.1units)for 10min to introduce nicks in the genomic DNA.The percentage of cardiomyocytes with DNA nick-end labeling was determined by counting the number of cells with brown-stained nuclei among1000nuclei in triplicate plates.2.9.Statistical analysisData are presented as means±standard error of the mean(SEM).Data for cultured cardiomyocytes are mean values of at least three different experiments. Student’s t-test for unpaired observations or one-way analysis of variance was used to compare data.P<0.05was considered statistically significant.Y.-Y.Wang et al./Molecular and Cellular Endocrinology 320 (2010) 67–7569Table 1General parameters in hyperthyroid and control rats.Group Heart rate (beats/min)SBP (mmHg)BM (g)HM (mg)HM/BMControl 341±19111±11354±10862.0±20.8 2.57±0.062-week T 4478±19*110±12316±15*1045.4±62.0* 3.31±0.20*4-week T 4542±17**,#108±12292±13*1312.0±30.5**4.50±0.20**Values are means ±SEM,n =6.SBP,systolic blood pressure.BM,body mass.HM,heart mass.2-or 4-week T 4,hyperthyroid for 2or 4weeks.*P <0.05vs.control values.**P <0.01vs.control values.#P <0.05vs.values in 2-week T 4-treated group.3.Results3.1.Heart rate,heart mass and cardiomyocyte sizeSince there were no significant differences between the 2-and 4-week synchronous control groups for size and functional param-eters,the data were combined as one control group.The heart rate was significantly increased by 40%(P <0.05)and 59%(P <0.01)in the 2-and 4-week T 4-treated groups,respectively,compared with the control group.The heart rate was 13%higher in the 4-week T 4-treated group than in the 2-week T 4-treated group (P <0.05).There were no differences in systolic pressure among the three groups.T 4-treated rats lost 11–17%of body mass compared with control rats.The absolute heart wet mass and ratio of heart mass to body mass were increased significantly in both T 4-treated groups relative to the control group (P <0.05,Table 1).The length,width and surface area of cardiomyocytes are sum-marized in Table 2.T 4treatment increased the width and surface area of cardiomyocytes,but not length.The width and surface area of cardiomyocytes in the 4-week T 4-treated group were greater than those in the 2-week T 4-treated group.3.2.Necrotic area,apoptotic rates,and DNA ladder in myocardiumThe right and left ventricular myocardium showed a normal structure in the 2-week T 4-treated group.Focal necrotic area was found in the right ventricular outflow tract (Fig.1),but not in the left ventricle,in 4-week T 4-treated rat hearts.Table 2Size of cardiomyocytes in hyperthyroid and control rats.Group Width (m)Length (m)Area (m 2)Control LV 21.0±1.2124.9±4.12753.8±196.5RV 22.5±0.8127.1±3.72802.1±215.22-week T 4LV 23.5±0.9*126.8±2.42993.8±138.0*RV 25.7±1.1*127.7±3.03196.7±147.1*4-week T 4LV 25.5±0.9*,#126.8±2.43187.6±138.0*,#RV27.7±1.0*,#129.0±2.73493.6±176.1*,#Values are means ±SEM,n =15cardiomyocytes per heart from 6rats of 2-or 4-week hyperthyroid.LV,left ventricle.RV,right ventricle.*P <0.05vs.control values.#P <0.05vs.values in 2-week T 4-treated group.Fig.2A,B and C shows TUNEL-positive myocyte nuclei in control,and 2-and 4-week T 4-treated hearts.The percent-age of TUNEL-positive nuclei was 1.4±0.2%and 1.6±0.3%in the control and 2-week T 4-treated hearts,respectively,with no difference between the two groups (Fig.2E).The apop-totic rate was significantly increased in the 4-week T 4-treated hearts compared with the synchronous control (P <0.01).DNA laddering was only detectable in the right and left ventri-cles from 4-week T 4-treated hearts (Fig.2D).Data shown in Figs.1and 2A,B,C,and D are representative experiments for six rat hearts.Fig.1.Epicardium of rat cardiac ventricles with HE staining.Representative images showing focal necrotic area (C,arrowheads)in the right ventricular outflow tract.Data are representative experiments of six rat hearts.(A)Control;(B)2-week T 4-treated heart;(C)4-week T 4-treated heart;(D)delineation of the observed sites.Scale bar,50m.70Y.-Y.Wang et al./Molecular and Cellular Endocrinology320 (2010) 67–75Fig.2.Assessment of cardiomyocyte apoptosis using an antibody against myosin heavy chain in conjunction with TUNEL staining.Representative immunofluorescent images showing apoptotic cardiomyocytes in control,and 2-and 4-week T 4-treated hearts.Scale bar,50m.Bar graphs (E)showing the average number of cardiomyocytes undergoing apoptosis.Values are means ±SEM,n =6hearts per group.**P <0.01vs.control.(A)Control heart and (B)2-week T 4-treated heart.Arrowheads indicate apoptotic myocyte nuclei.(C)4-week T 4-treated heart.Arrowheads indicate apoptotic myocyte nuclei.(D)Genomic DNA laddering assay.Data are representative experiments of six rat hearts.Electrophoretic pattern of low molecular weight DNA fragments extracted from freshly isolated left ventricular myocardium.CON,control.2-and 4-week T 4,2-and 4-week T 4-treated heart.RV and LV,right and left ventricle.Markers show 200-bp multiples.3.3.Cardiac output and stroke volumeThe intrinsic heart rate of the isolated working heart was increased in 2-and 4-week T 4-treated rats (P <0.01,Fig.3A).The cardiac output–preload relationship was positive in the control and both T 4-treated groups (Fig.3B).Cardiac output was increased at preloads of 5,10or 15mmHg in the 2-week T 4-treated group than in the control group,but there was no difference between the 4-week T 4-treated and control groups.The calculated stroke volume was unchanged at preloads of 5,10or 15mmHg in the 2-week T 4-treated group and was significantly lower in the 4-week T 4-treated group than in the control group (P <0.05or 0.01,Fig.3C).3.4.Frequency-dependence of shortening amplitude and contractile duration of unloaded contraction in isolated cardiomyocytesIsolated cardiomyocytes were stimulated at frequencies of 1,2and 4Hz.The percent shortening amplitude of unloaded contrac-tion increased with increasing frequency in the control group,but decreased in the 2-and 4-week T 4-treated groups (Fig.4).The per-cent shortening amplitude was higher in the 4-week T 4-treated group than in the 2-week T 4-treated and the control group at each frequency (P <0.01,Fig.4),but was only higher in the 2-week T 4-treated group than in the control group at frequencies of 1and 2Hz (P <0.01,Fig.4).The time to peak shortening and the time from peak to 75%relaxation of unloaded contraction were decreased at frequencies of 1,2and 4Hz in the 2-week T 4-treated group and were fur-ther decreased in the 4-week T 4-treated group compared with the control group (Fig.5A and B).The shortening and relaxation rates for unloaded contraction were increased at 1,2and 4Hz in the 2-week T 4-treated group compared with the control group.The shortening and relaxation rates showed a further increase at 2Hz but were reduced at 4Hz in the 4-week T 4-treated group (Fig.5C and D).3.5.Surface area and apoptotic rate in cultured cardiomyocytes Cultured cardiomyocytes were treated with T 3for 48h to induce dose-dependent hypertrophy.The surface area of untreated cardiomyocytes was defined as 100%.The surface area of cardiomy-Y.-Y.Wang et al./Molecular and Cellular Endocrinology320 (2010) 67–7571Fig.3.Cardiac output–preload relationship in working heart mode in hyperthy-roid and control rats.Since there were no differences between2-and4-week synchronous control groups for size and functional parameters,the data were com-bined as one control group.(A)Intrinsic heart rate;(B)cardiac output at different preloads;(C)calculated stroke volume.Values are means±SEM,n=6hearts per group.*P<0.05or**P<0.01vs.control values.ocytes reached123±3%,149±5%and151±6%with1,10and 100nM T3,respectively(P<0.01,Fig.6).The apoptotic rate of cardiomyocytes was2.6%in the control group and was2.4%and2.7%after treatment with1and10nM T3, respectively.There were no significant differences among thethree Fig. 4.Shortening amplitude–frequency relationship in cardiomyocytes from hyperthyroid and control rats.The percentage of shortening of unloaded single cardiomyocyte showing cardiac contractility.Values are means±SEM,n=10car-diomyocytes per heart from six rats per group(60cardiomyocytes per group). **P<0.01vs.control values.##P<0.01vs.2-week T4-treated group.groups.Treatment with1nM Ang II for24h increased the apoptotic rate to6.6%in control cardiomyocytes,and8.7%and10.9%in hyper-trophied cardiomyocytes induced by1and10nM T3treatment, respectively(Fig.7).4.DiscussionTetraiodothyronine(thyroxine,T4)(∼85%)and triiodothyronine (T3)(∼15%)are primarily synthesized and secreted by the thy-roid gland.T4,the primary secretary product,is relatively inactive. T3,the physiologically active form,is derived mainly from the peripheral conversion of T4in the skeletal muscle,liver,and kid-ney(Gomberg-Maitland and Frishman,1998).T3and T4are known to affect the development and function of almost every cell and organ,and exert very pronounced effects on the cardiovascular sys-tem.Excessive TH produces many cardiovascular manifestations including increased cardiac contractility and heart rate and cardiac hypertrophy in humans and animal models(Klein and Danzi,2007). Severe hyperthyroidism is also associated with heart failure(Klein and Ojamaa,2001).Although elevated levels of T3/T4directly or indirectly induce cardiac hypertrophy,the mechanisms involved in the transition from cardiac hypertrophy to heart failure remain unclear.To elucidate the mechanisms of heart failure,a rat model with hyperthyroidism was established by daily intramuscular or intraperitoneal injection or oral administration of thyroxine(Hu et al.,2003).This is a commonly used animal model.In our prelim-inary study,rats treated orally with200g/kg thyroxine per day induced heart failure within3weeks and had lower body mass, indicating significant stress was placed on the rats.Thus,we used the dose of100g/kg per day to induce hyperthyroidism in the present study.4.1.Thyroxine-induced hypertrophied cardiomyocytes are more susceptible to apoptotic stimulatorGerdes et al.(1985)reported that hyperthyroidism induced multifocal areas offibrosis in the right ventricle of rats. However,the mechanisms have remained unclear until now. Hyperthyroidism-induced myocardial hypertrophy is more pro-nounced in the right ventricle than in the left ventricle. Furthermore,hypertrophy is more marked in cardiomyocytes in72Y.-Y.Wang et al./Molecular and Cellular Endocrinology320 (2010) 67–75Fig.5.Contraction duration,rate of shortening and relaxation in unloading contraction of cardiomyocytes in hyperthyroid and control rats.(A)Time to peak shortening representing contraction duration;(B)time from peak to 75%relaxation representing relaxation duration;(C)shortening rate;(D)relaxation rate.Values are means ±SEM,n =10cardiomyocytes per heart from six rats per group (60cardiomyocytes per group).*P <0.05or **P <0.01vs.control values.##P <0.01vs.2-week T 4-treated group.the epicardium than those in the endocardium (Campbell and Gerdes,1988).Myocardium necrosis caused by thyroxine-induced ischemia and/or hypoxia can be excluded because the density of capillaries in the epimyocardium is greater than that in endo-cardium (White et al.,1988).If excessive TH directly affects the myocardium,it should cause widespread necrosis in the heart.However,necrosis was limited to focal areas in the right ventricle.Multifocal areas of fibrosis were also observed in hyperthyroid rats after treatment with propranolol.This indicates that -adrenergic stimulation is not necessary for thyroid-induced myocardial necro-sis (Gerdes et al.,1985).In the present study,the rat heart treated with T 4for 4weeks showed focal areas of necrosis beneath the epimyocardium of the right ventricle (Fig.1)and genomic DNA ladders in both right and left ventricles (Fig.2).Because car-diomyocytes account for 75–85%of heart mass,apoptosis might be derived from cardiomyocytes.Therefore,the fibrotic areas might result from apoptosis of cardiomyocytes in the right ventricle.Although few studies have evaluated T 4-induced apoptosis in the myocardium,excessive TH could lead to apoptosis in liver mediated by the mitochondrial pathway (Upadhyay et al.,2004)and in pan-creatic cells (Jorns et al.,2002).Excessive T 4was shown to activate the caspase pathway to induce apoptosis in Hela cells (Yamada-Okabe et al.,2003).In neonatal rat cardiomyocytes,T 3inhibited the expression of PKC (Rybin and Steinberg,1996),which is an important factor that protects against apoptosis in cardiomyocytes (Dorn and Brown,1999).Recently,Barreto-Chaves et al.reported that T 4increased the activity and expression of angiotensin-converting enzyme and elevated the serum Ang II concentration (Carneiro-Ramos et al.,2006).Ravassa et al.(2000)reported thathypertrophied cardiomyocytes had an increased susceptibility to Ang II-induced apoptosis.Ang II stimulated apoptosis of cardiomy-ocytes via the AT 1receptor and was associated with translocation of PKC and PKC ␦,which was coupled with an increase in intracel-lular Ca 2+in cardiomyocytes (Kajstura et al.,1997).In the present study we found that T 3induced dose-dependent hypertrophy in neonatal rat cardiomyocytes and that Ang II increased the apoptotic rate in hypertrophied cardiomyocytes.The apoptotic rate of car-diomyocytes was dependent on the extent of hypertrophy (Fig.7).Therefore,the hyperthyroidism-induced increased local Ang II con-centration and enhanced sensitivity to Ang II may increase the apoptotic rate in hypertrophied myocardium.Since apoptosis only occurred in the 4-week T 4-treated hearts,the total number of apop-totic cardiomyocytes may not be sufficient to induce heart failure in these hyperthyroid-treated rats.4.2.Cardiac contractility of hypertrophied cardiomyocytes is decreased in hyperthyroidism ratsAccording to the Frank–Starling law,cardiac output is depen-dent on preload or ventricular end-diastolic volume.Cardiac output increased with elevated preload in isolated working hearts from control,2-and 4-week T 4-treated groups (Fig.1).This indicates that the cardiac output–preload relationship is preserved in hyper-thyroidism.Since the isolated working heart is not regulated by autonomic nerve or by direct effects of TH,the increased intrinsic heart rate indicates that T 4increases the automatic rhythmicity of the sinus node.Higher heart rate can rapidly enhance cardiac con-tractility in most mammals (Langer et al.,2003).Thus,the cardiac。
Democracy and economic growth in Sub-Saharan Africa

Empir Econ(2013)45:987–1008DOI10.1007/s00181-012-0633-xDemocracy and economic growth in Sub-SaharanAfrica:a panel data approachVishal Chandr JaunkyReceived:16July2011/Accepted:16July2012/Published online:12September2012©Springer-Verlag2012Abstract This paper studies the link between democracy and economic develop-ment for28countries of Sub-Saharan Africa for the period1980–2005in a panel data framework.A democracy index constructed from the Freedom House indices.A variety of panel data unit root and cointegration tests are applied.The variables are found to be integrated of order one and cointegrated.The Blundell–Bond system generalized methods-of-moments is employed to conduct a panel error-correction mechanism based causality test within a vector autoregressive structure.Economic growth is found to cause democracy in the short-run,while bidirectionality is uncov-ered in the long-run.In addition,the long-run coefficients are estimated through the panel fully modified ordinary least squares and dynamic ordinary least squares meth-ods.Democracy has a positive impact on GDP and vice versa.These results lend support to the virtuous cycle hypothesis.Keywords Democracy·GDP·SSA·Panel dataJEL Classification C33·O401IntroductionDespite its huge resource endowments,the Sub-Saharan Africa(SSA),which adds up to48of the54African countries,remains to date,the poorest region in the world. In2007,its gross domestic product(GDP)per capita was estimated to be only about $1,869(World Bank2009).The SSA has been severely marked by a long series of civil wars and political turmoil(Jézéquel2006).Democracy has been lacking and this V.C.Jaunky(B)Centre for Energy Policy and Economics,ETH Zürich,Zürichbergstrasse18,8032Zurich,Switzerlande-mail:vjaunky@ethz.ch988V.C.Jaunky Table1GDP and democracy trend,1980–2005Countries Real GDP(million$)Democracy index19801990200020051980–200519801990200020051980–2005Benin1084.31411.82254.82727.21738.71.53.06.06.03.9 Burkina Faso1100.61555.62610.93529.51972.32.52.54.04.03.0 Burundi559.4865.1709.1789.7749.41.51.52.04.01.8 Cameroon6338.88792.77990.312056.59277.92.02.02.02.02.0 Cen.Af.Rep.735.3814.6959.4914.8836.42.02.54.53.52.9Chad664.91105.71385.12776.41279.31.51.52.52.52.0 Congo,DR.7015.87659.54305.85238.66185.91.52.03.03.02.6 Congo,Rep.1746.42795.63219.93975.52919.92.02.01.52.01.7Cote d’Ivoire7727.48297.610417.010409.08961.32.53.02.52.02.6 Gabon3594.34298.55067.85523.04578.82.04.03.53.03.0 Gambia,The213.5304.5420.9508.5333.75.56.02.03.54.0 Ghana2639.93266.94977.56364.13852.15.52.55.56.53.7 Kenya7086.810557.312705.315160.410721.53.52.02.55.02.7 Liberia1390.9433.0560.9444.2699.32.01.02.54.02.5 Madagascar3098.73265.63877.64339.13308.92.04.05.05.04.0 Malawi999.81243.01743.51827.11359.91.51.55.04.03.1Mali1536.01630.12422.53294.11973.21.52.55.56.03.8 Mauritius1518.42679.04469.35474.63204.05.06.06.57.06.3Niger1523.41507.01798.42207.01636.91.52.54.05.02.8 Nigeria31451.834977.845983.660557.038045.55.53.04.04.03.2 Rwanda1457.01781.61810.92351.11701.12.02.01.52.51.9 Senegal2682.73463.34691.85891.13871.64.04.54.55.54.4 Sierra Leone935.31021.6633.81201.6913.63.02.53.54.53.0 South Africa95502.5110944.7132877.6160792.9117675.62.53.56.56.54.5 Swaziland554.01023.81388.71561.61052.53.02.52.52.02.5Togo964.21070.71329.11479.51121.91.52.03.02.52.3 Zambia2729.83027.53237.74088.73101.02.52.53.54.03.5 Zimbabwe4376.46733.77399.25618.16382.04.53.02.51.52.9 Source Computed.Note In connection with the Democracy index,countries whose ratings average between 1.0and2.9are Not Free,those between3.0and5.4are Partly Free,and those between5.5and7.0are Freehas also arguably led to low income levels in the region.As stated by Diamond(2005),“…Africa cannot develop without democracy and that democracy in Africa ultimately cannot be sustained without development.”Furthermore,he defines democracy“…as a system of government in which the people choose their leaders and representatives, and can replace them,in regular,free and fair elections.”As exposed in Table1,while several states of the SSA are still undemocratic(Not Free),for some,there has been a slow butfirm march towards democracy.From an economic perspective,the question of whether democracy affects economic growth is critical to policymakers who seek to promote greater political freedom to fuel economic development in the SSA.Democracy and economic growth in Sub-Saharan Africa989 This paper explores the linkage between democracy and economic growth for a sample of28countries of the SSA1over the period1980–2005.Two genera-tions of panel unit root and cointegration tests are conducted.These are followed by a panel vector error-correction mechanism(VECM)-based causality test,uti-lizing the system generalized methods-of-moments(GMM)technique.The long-run estimates are computed by means of the panel fully modified ordinary least squares(FMOLS)and dynamic ordinary least squares(DOLS)estimators.Democ-racy scores are obtained from the Freedom House website.Real GDP(constant 2000)data are obtained from the World Bank CD-ROM2008.Similar to Narayan et al.(2011),real GDP is used to capture economic growth.The democracy index is simply built by averaging of the sum of civil liberties and political rights indi-ces.2The political rights index shows how fair and meaningful elections are carried out,while the civil liberties index involves freedom of press,freedom of speech, freedom of religious belief,and the right to protest and organize.These scores are re-adjusted whereby a score of1means least free,whereas a score of7reflects most free.Freedom House data are quite popular and have been used extensively in the liter-ature.Such application has to do with a variety of studies examining the implications of economic freedom on economic growth(Hanke and Walters1997),political insti-tutions on the environment(Bhattarai and Hammig2001),financial development on corruption(Altunba¸s and Thornton2012),among others.In addition to the Free-dom House,other freedom indicators are compiled by the Fraser Institute,Heritage Foundation,International Institute for Management Development(IMD),and World Economic Forum.These are assessed and compared by Hanke and Walters(1997), who pinpoint to several limitations accruing during the construction of the indicators by those institutions.However,as argued by Hanson(2003),“…Freedom House’s renown as an arbiter of political freedom and civil liberties gives its index instant cred-ibility,which may explain the failure of Hanke and Walters to challenge it(p.642).”The Freedom House index does have its criticisms.According to Minier(1998), the subjectivity involved when building the index brings in some measure of error and bias.Democracy is a multifaceted theme and the index is argued to be based on a checklist which includes limits on suffrage,freedom of the press,and restrictions on individuals running for office.In addition,the overall ranking done by the Freedom House can be debated to be entirely impressionistic.The index also tends to force a seemingly continuous variable into a discrete ranking system.Nevertheless,as argued by Tavares and Wacziarg(2001),the index is fashioned with the intension of consis-tency across time and across countries and this makes it suitable to use in a panel data setting.The rest of the paper is set out as follows:Section2provides an overview of the theoretical and empirical literature involving the democracy–economic growth nexus. Section3presents the testing frameworks.Section4discusses the results.Section5 concludes.1The selection of countries is done purely on the availability of data.2The characteristic of each score is available online at /about/fiw1.php.990V.C.Jaunky 2Review of theoretical and empirical literatureSince the seminal work of Lipset(1959),a voluminous number of scholarly works analyzing the link between economic development and democracy has emerged.As stated by him,“perhaps the most widespread generalization linking political system to other aspects of society has been that democracy is related to the state of economic development,”where educated people are apt to“…believe in democratic values and support democratic practices(p.75).”On the word of Pennar et al.(1993),“…rising incomes atfirst go toward needed goods and investment,then later toward more and more of what economists call”luxury goods,”such as higher education.A more edu-cated population tends to demand political and civil rights,and so democratization begins.”Economic growth is therefore conducive to democracy(Huber et al.1993; Barro2002).The Lipset hypothesis will be supported if in the long-run,a change in economic growth causes a change in democracy and a rise in economic growth results in greater democracy.The impact of democracy on economic growth is less straightforward and has been a matter of much more controversy among scholars.Mixedfindings have been observed.3To illustrate this connection,Sirowy and Inkeles(1990)put forward three major perspectives.These are namely the“conflict,”“compatibility,”and“skeptical”ones.First,the conflict perspective views democracy as a big hurdle to economic growth.To experience economic expansion,policies inhibiting excessive increase in real wages and promotion of both national and foreign capital accumulation are required.Unless these policies are adopted,rapid growth of industrialization has a tendency to to be delayed.This in turn slows down the process of economic growth. Democratic governments are looked upon to be vote maximizers and they are more concerned about implementing myopic policies such as state benefits and welfare policies at the expense of accumulation.Democracy has thus a negative impact on economic growth(Tavares and Wacziarg2001;Heo et al.2008).In essence,the con-flict hypothesis will be supported if in the long-run,a change in democracy causes economic growth and a rise in democracy has a negative effect on economic growth.On the contrary,the compatibility perspective is incompatible with an authoritarian model where economic development is exclusively directed by a centralized body or dictator.As a consequence,democracy is considered to sustain equitable allocation of resources and power,reduce distributional conflicts,and support fundamental civil liberties and political rights.These are appropriate to create the necessary socio-polit-ico-economic conditions favorable to economic growth.The impact of democracy on economic growth is expected to be positive(Kurzman et al.2002;Ghosh and Gregoriou2009).Fundamentally,the compatibility hypothesis will be supported if in the long-run,a change in democracy causes economic growth and an increase in democracy has a positive effect on economic growth.3Kurzman et al.(2002)review47quantitative studies of the effect of democracy on economic growth. 19found a positive relationship between democracy and growth,6found a negative relationship,and10 reported no statistically significant relationship.7studies found a combination of positive and non-sig-nificant results,depending on the model used and the cases included;2found a combination of negative and non-significant results;2found mixed positive and negative results;and1(Barro1996)reported an inverted-U effect.Democracy and economic growth in Sub-Saharan Africa991 Finally,from the skeptical perspective,there is no systematic relationship between democracy and economic growth(Rodrik1997).With regard to this perspective,fac-tors such as the effectiveness of government policies,institutional maturity,and the coordination of government entities,etc.play a more significant role in economic performance than the presence or absence of democracy(Heo2010).For instance, Przeworski and Limongi(1993)find no conclusive effect of democracy on economic growth.As stated by them“…the impact of political regimes on growth is wide open for reflection and research(p.66).”The skeptical perspective will be supported if no causal link between democracy and economic growth exists.In addition,a symbi-otic link between democracy and economic growth can correspondingly exist(Bhalla 1997),with both having a positive effect on each other.This can be referred to the virtuous cycle hypothesis.The Lipset and compatibility hypotheses are assumed hold at the same time.The existing empirical literature4has largely revolved around correlation and regression techniques while the causal effect between democracy and economic growth has actually been overlooked.As discussed above,reverse causality is an obvious pos-sibility and this can lead to biased estimates.So far,only a few studies have tried to examine any causal link between those two variables and these are mainly based on time series analysis.Heo and Tan(2001)study such link for32developing coun-tries over the period1950–1982.Their results support a causal relationship running from economic growth to democracy for Costa Rica,Egypt,Guatemala,India,Israel, Mexico,Nicaragua,South Korea,Thailand,Uruguay,and Venezuela.Next,unidi-rectional causality running from democracy to growth is found for Bolivia,Burma, Colombia,Ecuador,El Salvador,Indonesia,Iran,Paraguay,Philippines,and South Africa.Bidirectional causality is stumbled upon for Chile,Dominican Republic,and Turkey.Theyfind no relationship between economic growth and democracy for Argen-tina,Brazil,Haiti,Honduras,Pakistan,Panama,Peru,and Sri Lanka.Using ECM techniques,a few studies have applied more sophisticated causality tests to examine any short-run or long-run effects.Narayan and Smyth(2006)find a bidirectional causal relationship between democracy and economic growth in the short-run.A unidirectional causal relationship running from democracy to economic growth is likewise discovered in the long-run for China over the period1972–1999. Narayan et al.(2011)examine similar link for30countries of SSA over the period 1972–2001.They use two democracy indices,viz.the Freedom House political rights index and the Legislative index of Electoral Competitiveness(LIEC).Theyfind sup-port for the Lipset hypothesis for Botswana and Niger with both datasets,for Chad with the Freedom House data and for Cote d’Ivoire and Gabon with the LIEC data. Support for the compatibility hypothesis is found for Botswana with the Freedom House data and for Madagascar,Rwanda,South Africa,and Swaziland with the LIEC data.Support for the conflict hypothesis is found for Gabon with the Freedom House 4A relevant study includes Burkhart and Lewis-Beck(1994)where they examine the causal link between economic development and economic growth for131countries over the period1972–1989.They locate unidirectional causality running from economic development to democracy.Campos and Nugent(2002) test whether a causal and negative long term relation exist between political instability and economic growth 98countries andfind no evidence of such relationship.992V.C.Jaunky data and Sierra Leone with the LIEC data.For most of the countries in their sample,the skeptical hypothesis is supported.One major problem with econometrics research is the lack of sufficient observationsover long periods in the context of the need to test for unit roots and cointegrationamongst the series.Failure to do so exposes the study to the criticism of spuriousresults.Time series tests can be subject to criticism of low power especially whenthe number of observations is relatively small.As claimed by Toda(1995)“ (100)observations are not sufficient to ensure good performance of the tests(p.79).”Onepotential solution to this problem is to employ panel data techniques.These allow fora significant increase in testing power relative to conventional time series methods(Coakley and Fuertes1997).This paper attempts to add new empirical evidence tothe literature by revisiting the democracy–economic nexus for the SSA within a paneldata framework.3Testing frameworkTo test for any causal effects between democracy and economic growth,a panelGranger-type causality test is constructed.Granger causality does not however meantrue causality.Such statistical concept is based on linear regression stochastic pro-cesses(Granger1969).It can only be interpreted as showing whether prior changesin one series add(or do not add)significantly to the explanation of the future valueof another series(Farr et al.1998).The Granger causality test requires variables to bestationary,as it can yield spurious results(Granger and Newbold1974).This problemcan be avoided by taking advantage of a VECM-based causality test.Some prelimi-nary testing such as unit root and cointegration tests are required before carrying outthe Engle–Granger residual-type causality approach.First,both variables have to benon-stationary and integrated of same order,such as order one,denoted as I(1),totest for cointegration(Engle and Granger1987).Macroeconomic variables tend to benon-stationary in nature.A series Y t is integrated order of d,i.e.,Y t∼I(d),if it were to be differenced by d times to become stationary.Cointegration between democracyand real GDP should be established before proceeding to the VECM-based causalitytest.To verify the order of integration,a battery of panel unit root tests will be performed.Inferences on a single test can lead to inappropriate conclusion about the order of inte-gration as none of the current panel unit root tests are devoid of statistical limitationsin terms of size and power properties.Thefirst generation tests are conceptualizedby Hadri(2000),Levin et al.(2002,LLC),Im et al.(2003,IPS),and Im et al.(2005,ILT).These tests do not explicitly control for cross-sectional dependence.Each panelis assumed to operate independently from each other.This condition is rather unrealis-tic.Recent second generation relaxes the assumption of cross-sectional independenceand account for any correlation among the panels.These tests are derived by Pesaran(2007);Chang and Song(2009)and Hadri and Kurozumi(2012).Unit root tests are usually worked out using two distinct specifications.One testincludes a constant term only and the other contains both a constant term and a timetrend.These tests can be sensitive to the inclusion or exclusion of a trend.There shouldDemocracy and economic growth in Sub-Saharan Africa993 be a careful use of a deterministic trend or“otherwise results can be misleading(p.51)”(Ahking2002).Macroeconomic data tend to exhibit a trend over time.Intuitively, it is more apt to consider a regression with a constant and a trend at level form.Since first-differencing tends to remove any deterministic trends in the variables,inferences will be done in line with a specification with a constant term only.For the sake of completeness,both specifications are computed.After checking the order of integration of the two variables,panel cointegration tests can be afterward conducted.First,Nyblom and Harvey(2000)propose a non-parametric cointegration test of common trends.The test assumes where H0is the stationarity of the series around a deterministic trend and there exists k<n common trends(where rank( η)=k),against the alternative of a random-walk component incidence,where there exists more than k common trends(where rank( η)>k).This test tests for the H0of0common trends against the hypothesis of common trends among the variables.The non-parametric cointegration test is followed by two parametric ones.Second,Pedroni(1999)proposes several tests with the H0of no cointegration.These computed statistics are called within-dimension-and between-dimension-based statistics.Similar to panel unit root tests,panel cointegration tests can suffer from the problems of cross-sectional dependence.Westerlund(2007)advo-cates a cointegration test which can handle this issue.This can be considered as a second-generation panel cointegration test.This test is based on structural instead of residual dynamics and there is no one common factor restriction.Baltagi(2008)pro-vides a thorough review of the panel unit root and cointegration literature which indeed points towards the vital importance of controlling for cross-sectional dependence.If the variables are cointegrated,causality should occur in at least one direction (Baffes and Shah1994).Consistent with Jaunky(2011),theρth order VECM struc-ture can be represented as follows:LGDP it LDEM it=α1α2+ρk=1β11kβ12kβ21kβ22kLGDP it−kLDEM it−k+φ1φ2ECM it−1+ε1itε2it(1)where i=1,...,N;t=ρ+1,ρ+2,...,T;theαs,βk s,andφs are parametersto be estimated.ECM it−1represents the one period lagged error-term derived fromthe cointegrating vector and the error termsε1it andε2it are assumed to be seriallyindependent with mean zero andfinite covariance matrix.Given the use of a vectorautoregressive(V AR)model,all variables are treated as being endogenous.LDEM itand LGDP it denote the natural logarithmic of real GDP and democracy for countryi ay time t.A simple Wald test for the joint significance can be applied to examinethe direction of any causal relationship.For instance,democracy does not Granger-cause economic growth if and only if all the coefficientsβ12k;∀=1,2,...,ρare not significantly different from zero in Eq.(1).The dependent variable reacts merely toshort-term shocks.In the similar fashion,economic growth does not Granger-causedemocracy in the short-run if and if all the coefficientsβ21k;∀=1,2,...,ρare not significant from zero.These can be referred to as the“short-run Granger causality”994V.C.Jaunky test.The coefficients on the ECM s represent how fast deviations from the long-run equilibrium are eliminated.An additional channel of causality can be studied by test-ing the significance of the ECM s.This test is referred to as the“long-run Granger causality”test.The estimators in Eq.(1)are biased due to the correlation among the lagged depen-dent variables and the error terms.To account for the correlation and endogeneity problems,Arellano and Bond(1991)suggest a two-step difference GMM approach, where the lags of explanatory variables in levels are to be used as instruments.In the first step,the error terms are assumed to be independent and homoscedastic.In the second step,thefirst-step residuals are applied to construct consistent variances and covariances matrices,with the former assumptions are relaxed.For the instruments to be valid,ε1andε2should not be serially uncorrelated.This condition occurs when there is statistical evidence of a significant and negativefirst-order serial correlation (AR(1))and no evidence of second-order serial correlation(AR(2))in the differenced residuals mutually.The optimal lag lengthρis chosen if such condition is satisfied.The Arellano–Bond estimator suffers from a lack of power of the internal instru-ments.Following Blundell and Bond(1998),if the lag of the dependent variable and the explanatory variables are persistent in time,lags of the levels of these variables will be weak instruments for the equation in differences.They consequently recommend the system GMM.This estimator is a linear combination of the levels and differences, where the weight specified to the levels estimators grows in the prevalence of weak instruments owing to the high persistency in the series.In the presence of heteroske-dasticity and serial correlation,the two-step system GMM employs of a consistent estimate of the weighting matrix,using the residuals from the one-step estimator (Davidson and MacKinnon2004).In contrast to the conventional OLS method,the system GMM does not assume normality and controls for heteroskedasticity.The two-step system GMM estimator is more efficient than the one-step one.Nev-ertheless,it converges slowly to its asymptotic distribution and its standard errors tend to be biased downwards forfinite samples.The one-step estimator does not experi-ence such problem.Thefinite-sample correction to the two-step covariance matrix as derived by Windmeijer(2005)can be employed.The superiority of the two-step robust system GMM can be maintained.This approach uses multiple lags as instruments.The system is as a consequence over-identified.To test if the model is correctly specified and instruments are valid,both the Hansen(1982)J and the Sargan(1958)tests are computed.The latter is not robust to heteroscedasticity or autocorrelation,whereas, the former,which is the minimized value of the two-step GMM criterion function,is. The system GMM makes an exogeneity assumption where any correlation between endogenous variables and unobserved orfixed effects are constant over time.This allows the inclusion of levels equations in the system and use of lagged differences as instruments for these levels.The exogeneity assumption can be tested by using a difference-in-Hansen test(Bond et al.2001).Besides,the long-run impacts are required to test the aforementioned hypotheses. Bidirectional causality between LDEM it and LGDP it is found in the long-run.This situation is synonymous to endogenous regressors which can produce inconsistent and biased estimates.Long-run and efficient estimates can be obtained through the FMOLS and DOLS.These respective non-parametric and parametric methods canDemocracy and economic growth in Sub-Saharan Africa995 Table2LLC panel unit root test statisticsVariables Deterministics Level form First-differencet value t*t value t*LGDP it Constant−5.811−1.50693[0.066]‡−12.581−1.47332[0.070]‡Constant+trend−10.204 1.43561[0.924]−14.055−0.23175[0.408] LDEM it Constant−6.662−0.15055[0.440]−16.121−2.57481[0.005]* Constant+trend−10.6370.60743[0.728]−17.5370.11423[0.5455] Source computed.Note These LLC statistics are distributed as standard normal as both N and T grow large. Assuming no cross-country correlation and T is the same for all countries,the normalized t*test statistic is computed by using the t value statistics.After transformation by factors provided by LLC,the t*tests is distributed standard normal under the H0of non-stationarity.It is compared to the1,5,and10%sig-nificance levels with the one-sided critical values of−2.326,−1.645,and−1.282correspondingly.The p values are in square brackets.*,**,and‡denote1,5,and10%significance level,respectively.These notations are applied for all succeeding testseffectively correct for the biases resulting from the endogeneity of regressors over and above serially correlation and heteroskedasticity in error terms(Pedroni2001). According to Kao and Chiang(2000),the DOLS method tends to outperform the FMOLS estimators in term of mean biases.Equation(1),in conjunction with the long-run estimators,can be used to test the hypotheses linking democracy and economic growth.The Lipset hypothesis holds if, in the long-run,real GDP Granger-causes democracy and an increase in real GDP has a positive effect on democracy.Subsequently,the compatibility hypothesis holds if, in the long-run,democracy Granger-causes real per capita GDP and an increase in democracy results in an improvement in real per capita GDP.In contrast,the conflict hypothesis holds if,in the long-run,democracy Granger-causes real per capita GDP while a rise in democracy has a negative effect on real GDP.The skeptical hypothesis holds if there is no causal relationship between democracy and real GDP.Finally,the virtuous cycle hypothesis holds if bidirectionality prevails and each variable having a positive effect on each other.4ResultsFor implementation of the panel unit root tests,the Bartlett kernel is used.All band-widths and lag lengths will be equal to4(T/100)2/9≈2.97,where T=26(Basher and Westerlund2008).The maximum lag length lies between2and3.Too few lags may adversely affect the size of a unit root test,while too many lags can reduce the power of the unit root test(Campbell and Perron1991).Martins(2011)applies2 lags when performing the Pesaran panel unit root test over a period of1980–2005. Since the time span is relatively short,the lowest value possible of2is chosen to conduct the panel unit root tests.The LLC test statistics are reported in Table2.Both LGDP it and LDEM it are computed to be non-stationary and I(1),in keeping with the above-mentioned intuition about the order of integration.。
全自动化学发光免疫分析仪Immulite2000标准操作规程

IMMULITE® 2000全自动化学发光免疫分析仪标准操作程序本文件仅供参考,各实验室需根据各自情况建立自己的操作规程IMMULITE®2000全自动化学发光免疫分析系统标准操作规程SOP编码:页数:制定人签名:日期:审核人签名:日期:批准人签名:日期:生效日期:颁发日期:周期性审查:年一次修订登记:审查登记:[目的]描述IMMULITE®2000全自动化学发光免疫分析系统的使用和维护。
[范围] 适用IMMULITE®2000全自动化学发光免疫分析系统的操作。
[仪器工作原理和检测过程]1 IMMULITE 2000使用包被特定抗体的聚苯乙烯珠子作为固相,包被珠放在一个特定的反应杯中,从而进行温育,清洗以及信号发生。
2. 样本与结合了碱性磷酸酶的试剂温育反应结合之后,通过高速离心将剩余试剂甩到与反应杯同轴的废液管路中。
系统在几秒钟内完成四次离心清洗,以便与系统的其他同步。
去除未包被试剂的包被珠仍然保留在反应杯中。
3. 包被珠上的结合标记随后同发光底物进行定量发光。
当包被珠上结合的碱性磷酸酶标记同化学发光底物反应时,就产生发光。
发光强度同样本中待测物的含量有关。
仪器通过光电倍增管检测发光强度,随后计算出每个样本的结果。
4. 操作者在IMMULITE 2000上运行测试时,需要做下列操作::4.1进行每日探针清洗工作。
4.2 选择RUN IMMULITE 2000按钮。
4.3 查系统状态指示,加满或者清空耗材或者废物。
4.4 初始化样本和试剂加样器、蒸馏水喷嘴和底物喷嘴。
4.5 使用图像扫描器扫描试剂转盘上的过敏原试剂楔。
4.6在样本转盘上装载病人血清、质控、校正液和稀释液。
注意: 运行仪器需要用到的试剂都在 IMMULITE 2000试剂盒中。
只有需要预稀释的测试项目才会有稀释液。
4.7 在工作列表列表界面为样本指定测试项目以及编号。
4.8检查试剂以及与之匹配的包被珠是否足够完成所需测试。
Guest Editors ’ Introduction Large-Format Displays

I magine a future when every surface in everybuilt space is a high-resolution active dis-play with sensing capabilities.Scenario: I’m walking down the hall toward my officewhen I’m reminded that I’m late for a meeting with Maryto discuss the design of the students’ center being built oncampus. Unsure of the location of her office, I tap on thewall next to me and a large floorplan appears. After fol-lowing a path displayed on the floor to guide me, I arriveat her office, and we begin to work. Mary and I view animmersive walkthrough of the design on her “smart wall,”and I draw modifications with a virtual marker, whosestrokes are recognized and used to manipulate the com-puter model …Scenarios like this one are no longer science fiction—the technologies to make interactive display surfacescommonplace are being invented and deployed rightnow. Soon we will move into a new era of computing—ubiquitous visual computing based on enormous visualinformation and natural user interactions. The reasonsfor this new era are two-fold: increased computer per-formance and developed display techonology.First, continuous rapid improvements in CPU perfor-mance, storage density, and network bandwidth haveprovided sufficient bandwidth and computationalresources to support high-resolution displays and nat-ural human-computer interactions. Nowadays, the mainbandwidth bottleneck in an interactive computer sys-tem occurs in the link between computer and human,not between computer components within the system.Second, large-format display devices, such as projec-tors and flat panels, are rapidly becoming commodityitems. Meanwhile, new display technologies, such asorganic light-emitting diodes (OLED), will soon becomeavailable at inexpensive prices. They can be attached toalmost any kind of surface, allowing unlimited freedomof design for the interiors and exteriors of rooms andbuildings.We believe that new display technologies will revo-lutionize the way we use computers, making us rethink the relationship between information technology and our society. As an example, consider how wall-sized dis-plays enable qualitatively different human-computer interactions than traditional desktop displays.A recent paper by Mayer divides viewing experience into four categories.1The first category is the “postage-stamp experience,” where the field of view is con-strained by desktop system bandwidth limitations and display technology limitations. The second is the “tele-vision experience.” Television is ubiquitous and it’s the baseline of multimedia applications on a digital com-puter. The third is the “theatrical experience,” where emotions are involved. A theater offers a large field of view to enable viewers to use eye scan motion to expe-rience the image. Yet, it’s constrained by the frame size.The last is the “immersive experience,” which is when the scope and resolution of the imagery become large enough that viewers can leave the center of their focus and turn both their head and their attention to discov-er and study details of the scene and contextual envi-ronment, such as in an iMax movie theater.Unfortunately, current desktop display devices lack the field of view to provide anything more than a “tele-vision experience.” In contrast, large-scale displays can immerse users’ entire fields of view in computer-gener-ated imagery. Their high resolution enables visualiza-tion of detailed data sets, their size creates the illusion of immersion, and the space in front of them supports natural interactive collaborations among multiple simultaneous viewers.Traditionally, large-format electronic displays have been used mostly for specialized, high-end applications such as flight simulators, scientific visualization, and mechanical CAD. However, recently, the software and hardware required to build large displays have dramat-ically improved in price and performance, and several research and commercial instituitions are developing new methods for building and using them. As lower cost 0272-1716/00/$10.00 © 2000 IEEEGuest Editors’ Introduction20July/August 2000Thomas Funkhouser and Kai Li Princeton University Large-FormatDisplaysIEEE Computer Graphics and Applications21。
Functional interaction of phytochrome B and cryptochrome 2

................................................................. Functional interaction of phytochrome B and cryptochrome2Paloma MaÂs,Paul F.Devlin,Satchidananda Panda&Steve A.KayDepartment of Cell Biology and National Science Foundation Center for Biological Timing,The Scripps Research Institute,10550North Torrey Pines Road,La Jolla, California92037,USA .............................................................................................................................................. Light is a crucial environmental signal that controls many photo-morphogenic and circadian responses in plants1.Perception and transduction of light is achieved by at least two principal groups of photoreceptors,phytochromes and cryptochromes2,3.Phyto-chromes are red/far-red light-absorbing receptors encoded by a gene family of®ve members(phyA to phyE)2,4in Arabidopsis. Cryptochrome1(cry1),cryptochrome2(cry2)and phototropin are the blue/ultraviolet-A light receptors that have been charac-terized in Arabidopsis5.Previous studies showed that modulation of many physiological responses in plants is achieved by genetic interactions between different photoreceptors6;however,little is known about the nature of these interactions and their roles in the signal transduction pathway.Here we show the genetic inter-action that occurs between the Arabidopsis photoreceptors phyB and cry2in the control of¯owering time,hypocotyl elongation and circadian period by the clock.PhyB interacts directly with cry2as observed in co-immunoprecipitation experiments with transgenic Arabidopsis plants overexpressing ing¯uor-escent resonance energy transfer microscopy,we show that phyB and cry2interact in nuclear speckles that are formed in a light-dependent fashion.Light signals markedly affect the pace of the clock by activating different photoreceptors7.In response to increasing light intensity, the period length of the circadian expressed CAB2::luciferase gene8 decreases7.Here,cry2mutant seedlings displayed a de®ciency in the perception of white light,as shown by a longer period length of CAB2::luciferase expression at intermediate¯uence rates(Fig.1a). This effect was not observed in red light(Fig.1b),whereas in blue light only a small increment was observed at high¯uence rates (Fig.1c).These results indicate that full cry2function is only apparent on simultaneous illumination with multiple wavelengths of light,and suggests that it has a requirement for phytochrome activation.The signi®cance of the shorter period length in cry2 mutants at the lowest red or blue¯uence rates remains unclear. Light with a reduced red/far-red(R/FR)ratio that mimics light re¯ected from neighbouring vegetation causes the conversion of phytochrome from the active Pfr form to the inactive Pr form,and results in a shade-avoidance response including promotion of elongation growth and accelerated¯owering9.Cry2-de®cientFigure1Physiological and biochemical evidence for interaction between cry2and phyB. a±c,Effect of light intensity on mean period length(6s.e.)of the circadian rhythm of CAB2::LUC bioluminescence in wild type and cry2seedlings in white light(a),red light (b)or blue light(c).Asterisk,P,0.01(Student's two tail heteroscedastic t-test).d,Mean ¯owering time(6s.e.)in wild-type and cry2seedlings in high(white)versus low(white +FR)R/FR ratio.e,Mean hypocotyl length(6s.e.)in wild-type and cry2seedlings in high(white)versus low(white+FR)R/FR ratio.f,Western blot analysis of co-immunoprecipitation(co-IP)experiments with anti-phyB antibody and detection with antibody to cry2.The membrane was stripped and reprobed with anti-phyB antibody. `Total'indicates total protein extracts not subjected to immunoprecipitation.cry2+,cry2 overexpression Arabidopsis lines;phyB,phyB mutant Arabidopsis lines.-Ab indicates samples processed without incubation with phyB antibody.Arabidopsis plants have a delayed¯owering phenotype in response to extended photoperiods10,and regulation of¯oral induction is mediated by the antagonistic actions of cry2and phyB11.Here,the late¯owering phenotype of cry2in white light was no longer apparent under a low R/FR ratio(white+FR light),showing a marked acceleration of¯owering time(Fig.1d).These results indicate that the active Pfr form of phytochrome is required for the expression of cry2phenotype.Thus,either cry2suppresses phyB signalling,or removal of Pfr overrides other pathways in which cry2 regulates¯owering.Furthermore,although cry2seedlings showed a wild-type hypocotyl length in white light,they displayed a long hypocotyl phenotype in white+FR light(Fig.1e).We propose that long hypocotyl phenotype of cry2is not apparent in white light because it is negated by increased phyB action in the absence of cry2 (phyB is the major phytochrome that inhibits hypocotyl elongation in white light12).In agreement with this,cry2seedlings showed a greater promotion of elongation growth than wild-type seedlings. These results are consistent with a suppression of phyB signalling by cry2.Alternatively,a conditional redundancy between cry2and phytochrome signalling may also be possible.To examine whether phyB directly interacts with cry2,we performed co-immunoprecipitation assays with Arabidopsis trans-genic plants overexpressing cry2(cry2+)13.Immunoprecipitation with anti-phyB antibody and subsequent detection of cry2(Fig.1f, anti-cry2western blot)showed a band with a relative molecular mass of about75,000(M r<75K)(lane co-IP,cry2+)coincident with the size of the band observed in total protein extracts not subjected to immunoprecipitation(lane total,cry2+).The absence of signal in phyB-de®cient Arabidopsis mutant plants(lane co-IP, phyB)or in samples processed without anti-phyB antibody(lane ±Ab,cry2+)indicated the speci®city of the interaction.Reprobing the same blot with anti-phyB antibody(anti-phyB western blot) con®rmed phyB immunoprecipitation(lane co-IP,cry2+)and the absence of signals in negative controls(lanes co-IP,phyB and cry2+, -Ab).These results are consistent with a direct,physical interaction between phyB and cry2in Arabidopsis plants.Previous reports described the nuclear localization of Arabidopsis cry2(refs14,15).Both human and mouse cry2also localize in the nucleus3suggesting that the nuclear localization is important for cry2function.We therefore examined the effects of light in cry2 subcellular localization using a fusion protein of cry2with the red ¯uorescent protein(RFP).In transfected BY-2protoplasts main-tained in the dark,cry2±RFP appeared homogeneously distributed within the nucleus(Fig.2a).Rapid scanning with the blue laser lineFigure2Effects of light on cry2±RFP subcellular distribution and colocalization with phyB±GFP in BY-2cells.a,In the dark,cry2±RFP appeared distributed homogeneously in the nucleus.b,Formation of nuclear speckles after successive scanning with the blue laser of the confocal microscope.c,Superimposition of bright-®eld and¯uorescent images.d,e,Nucleus of a cell co-expressing phyB±GFP(d)and cry2±RFP(e)examined after10h in continuous red light and scanning with the488-nm laser line of the confocal microscope for phyB(d)and cry2(e)speckle formation.f,phyB±GFP and cry2±RFP colocalization after superimposing the images.g±o,phyB±GFP(g,j,m)and cry2±RFP (h,k,n)nuclear speckles examined at30-s intervals in the same nucleus.Yellow in merged images(i,l,o)indicates sites where phyB and cry2colocalize.Scale bar,5m m (a±c);2m m(d±o).Figure3FRET analysis of phyB±GFP and cry2±RFP interaction.a,Spectral overlap (J da)between the GFP emission and RFP excitation ing a value of22,500M-1cm-1for the molar extinction coef®cient(e a)of RFP,J da was calculated as 1.67´10-13M-1cm3.From this value and a quantum yield(f d)of0.60for GFP,a refractive index(n)of1.4and a dipole orientation factor(k2)of2/3,we calculated a critical transfer distance(R0)of5nm.b,BY-2cells transfected with phyB±GFP examined with the488-nm laser of a confocal microscope using a FVX-BA550RIF®lter.c,Background signal of the red channel examined with a568-nm krypton laser and using anFVX-BA585IF®lter.e,Adjustment of the threshold settings and background subtraction to eliminate crosstalk between the channels.f,g,i,Cells co-transfected with phyB±GFP(f) and cry2±RFP(g)excited with the488-nm laser and observed using an FVX-BA585IF ®lter(i).d,h,Merged images of green and red channels.Scale bar,2m m.of the confocal microscope or irradiation with blue light induced the formation of nuclear speckles(Fig.2b).Superimposition of bright-®eld and¯uorescent images clearly revealed the nuclear accumulation of cry2(Fig.2c).Cry2±RFP did not induce speckles in cells maintained in the dark,indicating that light(not over-expressed proteins)may be responsible for the formation of these speckles.Cry2±GFP fusion protein also localized in nuclear speckles,indicating that these speckles were not due to artefactual expression of RFP in plants.Fluorescent speckles were never observed when protoplasts were transfected with non-fused RFP or RFP fused with cry1(data not shown).Previous studies reported that phyB translocates to nuclear speckles under continuous red light16,17.To examine the subcellular relationship between phyB and cry2,we co-transfected BY-2 protoplasts with phyB±GFP and cry2±RFP,and examined the ¯uorescent signal by confocal microscopy.Co-transfected proto-plasts were maintained8h in the dark followed by irradiation with continuous red light for accumulation of phyB±GFP in nuclear speckles(Fig.2d).These cells were then irradiated with blue light or scanned with the blue laser of the confocal microscope to induce cry2speckles(Fig.2e).Superimposing both images revealed that some of the speckles where cry2and phyB accumulated perfectly colocalized in the nucleus(yellow stain in Fig.2f).PhyB and cry2 colocalize in some but not all of these nuclear speckles.It is possibleFigure4FRET microscopy by acceptor photobleaching.a,b,Fluorescence intensity of phyB±GFP in a demarcated region(yellow circle)was measured before and after cry2±RFP photobleaching in samples where phyB±GFP and cry2±RFP colocalized(a),or did not colocalize(b).c,Fluorescent intensities of GFP before and after RFP photobleaching were also compared in samples where non-fused GFP and RFP colocalized.Results consistent with those shown here were obtained in®ve independent experiments.that both photoreceptors might also have independent functions and that they recruit,in these speckles,other nuclear factors involved in light responses.We next investigated the changes of phyB±GFP and cry2±RFP distribution over time in protoplasts immobilized in agarose and examined by confocal microscopy at 30s intervals.Figure2g±i depicts an optical section of a nucleus showing phyB±GFP and cry2±RFP distribution in speckles that did not colocalize(row0).Scanning at30-s intervals the same cell revealed that some(row30s)or most(row60s)of cry2speckles did colocalize with phyB(yellow stain in merged image).Together,these results demonstrate the light-dependent colocalization of phyB and cry2in speci®c nuclear speckles.We next used¯uorescent resonance energy transfer(FRET) microscopy to determine whether the nuclear speckles are the sites where phyB and cry2interact.FRET is a quantum mechanical process by which a¯uorescent molecule,the donor,transfers energy by a non-radiative mechanism to an appropriately positioned chromophore,the acceptor18.Successful methods for the detection of intramolecular FRET signals in living plant cells have been developed using chromophore mutants of GFP19.We report,for the®rst time to our knowledge,the use of GFP±RFP combination for intermolecular detection of FRET signals in living cells.Before FRETanalysis,we calculated GFP±RFP spectral overlap(J da=1.67´10-13M-1cm3)and FoÈrster distance(R0=5nm)to show that GFP±RFP combination is a suitable donor±acceptor pair for FRET (Fig.3a).The rationale of our experiments resides in the fact that FRET would be detected only when the¯uorophores are directly interacting and not simply colocalizing.We®rst calibrated the microscope system using cells transfected only with phyB±GFP (Fig.3b)and no cry2±RFP(Fig.3c).The samples were used to subtract the background and to adjust appropriately the threshold settings(Fig.3e).This control veri®ed that the signals obtained in FRETwere not due to channel overlap.To measure FRET,we excited cells co-expressing phyB±GFP(Fig.3f)and cry2±RFP(Fig.3g) with the488nm laser-line,and observed cry2±RFP emission using a FVX-BA585IF®lter.Signi®cant FRET emission of cry2±RFP (Fig.3i,FRET)was observed in speckles that colocalized with phyB±GFP(see yellow in merged image,Fig.3h).These results indicate the direct molecular interaction of phyB and cry2in nuclear speckles and con®rm the previous results obtained in co-immunoprecipitation experiments.FRET measurement as described above can lead to errors,mainly owing to differences in the concentration of phyB±GFP and cry2±RFP or to the direct excitation of RFP at the GFP excitation wavelength.Furthermore,background subtraction to minimum ¯uorescence values(Fig.3e)might mask less strong FRET signals and eliminate the possibility of obtaining FRETef®ciency maps.We therefore evaluated FRET using a different method that is based on the increase of the donor¯uorescence after photochemical destruc-tion of the acceptor20.Cells co-expressing phyB±GFP and cry2±RFP (Fig.4a)were maintained in continuous red light and imaged,®rst with the488-nm laser line(phyB±GFP)and later with the568-nm laser line(cry2±RFP).We chose one of the speckles where phyB±GFP(green)and cry2±RFP(red)colocalized(yellow in merged image)and measured the¯uorescence intensity of phyB±GFP in a region roughly4m m in diameter(demarcated by a yellow circle; Fig.4a,pre-bleach).Cry2±RFP was subsequently photobleached by repeated scanning with the568-nm laser line until no signal was detected(cry2±RFP,Fig.4a post-bleach).We then measured the ¯uorescent intensity of phyB±GFP and compared it with that obtained in the same region before bleaching.A clear increase of phyB±GFP¯uorescence was observed(compare pre-and post-bleach in Fig.4a).This effect was not detected in phyB±GFP speckles that did not colocalize with cry2±RFP(Fig.4b).In those cases,the levels of¯uorescence intensity of phyB±GFP after cry2±RFP bleaching remained the same or slightly decreased as compared with those observed before bleaching(pre-and post-bleach in Fig.4b).To verify that FRET signals were not due to¯uorophore colocalization,we carried out an additional negative control with cells co-expressing non-fused RFP and non-fused GFP constructs. Bleaching RFP that colocalized with GFP did not increase GFP ¯uorescent intensity relative to its¯uorescence before bleaching (Fig.4c,pre-and post-bleach).Our results show that phyB±GFP ¯uorescence increased at sites where FRETwas occurring,con®rming the physical interaction between cry2and phyB in nuclear speckles. The light-induced nuclear compartmentalization of phyto-chromes16,17,the discovery that they possess kinase activity21and the identi®cation of some of their interacting partners22±24have provided crucial understanding of how phytochromes transmit light signals.These®ndings suggest that regulation of protein expression,phosphorylation and subcellular compartmentalization may have key roles in mediating light-induced physiological responses in plants.Cry1and cry2are phosphorylated by the kinase activity of phyA25.Our results show that phyB and cry2 interact in speci®c nuclear speckles.It is possible that these speckles act as`transcriptosomes'similarly to some factors described in animal cells that accumulate in a characteristic speckled pattern and are involved in processing and transcription of RNA26.Light-induced responses are also mediated by alteration of protein stability.Cry2is strongly downregulated by blue-light13,so the nuclear speckles could represent sites where cry2is degraded. Recent studies have suggested that novel clock-associated compo-nents contain domains involved in targeting speci®c substrates for proteolytic degradation27,28.Further experiments need to clarify whether these clock components are involved in cry2degradation, and whether phyB has a role in this process.M MethodsMeasurements of growth and¯oweringWild-type and cry2seeds were grown for6d in constant white light(60m mol m-2s-1)at 228C before being either maintained in constant white light(R/FR ratio8.53)or transferred to white light supplemented with FR(R/FR ratio0.05).A low R/FR ratio light was generated by supplementing white light with FR light from two LED light sources (Quantum devices,Barneveld,WI).Light measurements were made using an LI1800/12 spectroradiometer(Li-Cor,Lincoln,NE).Flowering time was measured as the number of rosette leaves at bolting(production of a1-cm high cauline stem).Data represent the mean(6s.e.)of6±8plants for each treatment.Hypocotyl length was measured after10d in the treatments using Scion Image software.Data represent the mean(6s.e.)of28±31 seedlings for each treatment.Light input to the circadian clockWild-type and cry2seeds expressing a CAB2::LUCIFERASE reporter construct8were grown in12h white light/12h dark cycles for6d before being transferred to constant red light(600±700nm),blue light(400±500nm)or white light at the intensities indicated. The rhythm of bioluminescence,representing CAB2transcription,was followed as described7and period length was calculated by®tting a cosine wave function to the time series for each seedling7.Data represent the mean(6s.e.)of5±18seedlings,representative of2±4independent experiments.Coimmunoprecipitation experimentsCry2overexpression plants13(provided by C.Lin,Univ.California,Los Angeles)were grown in12h white light/12h dark cycles and processed for co-immunoprecipitation in the®rst15min of the light cycle.PhyB immunoprecipitation was performed at48C for2h using anti-phyB antibody(provided by A.Nagatani,Kyoto Univ.,Kyoto)in a buffer containing10mM HEPES pH7.5,10%glycerol,100mM NaCl,0.2%Triton X-100,1mM EDTA and1mM dithiothreitol with a mixture of protease inhibitors(Sigma).Gamma-bind plus sepharose beads(Pharmacia)were used to precipitate the immunoprotein complexes.Cry2detection was performed using anti-cry2antibody(provided by C.Lin, Univ.California,Los Angeles).Plasmid construction and confocal imagingRed¯uorescent protein(RFP)coding region was obtained from the vector pDsRed1-N1 (Clontech Laboratories).Cry2coding sequence was obtained from C.Lin.The fusion protein cry2±RFP was cloned into the vector pRTL2which contains a CaMV35S promoter29.Colocalization of phyB±GFP16and cry2±RFP was examined in BY-2proto-plasts prepared and transfected as described30.Protoplasts were maintained8h in the dark followed by irradiation with continuous red light(15m mol m-2s-1)for10h.Cry2speckle formation was induced after irradiation with blue light(5m mol m-2s-1)for1min or after successive scanning with the488-nm laser of a confocal laser-scanning microscope(IX70; Olympus Corp).To examine GFP¯uorescence,we scanned samples with a488-nm argonlaser line using a550-nm(FVX-BA550RIF)barrier®lter and´601.4NA PlanApo oil-immersion objective.RFP signal was examined with a568-nm krypton laser using an FVX-BA585IF®lter.FRET analysisNormalized¯uorescence emission spectrum of GFP(F d)(provided by G.Patterson and D.Piston,Vanderbilt Univ.,Nashville,Tennessee)and absorption spectrum of DsRFP (provided by Clontech)were used to calculate GFP±RFP spectral overlap(J da)and FoÈrster distance(R0)according to the equations18:J da e`0F d l e l l4d le`0F d l d l1andR60 8:75310225 k2n24f d J da 2 where e is the molar extinction coef®cient of RFP;f d is the quantum yield for GFP;n is the refractive index;and k2is the dipole orientation factor.FRET signals were examined by excitation with the488-nm argon laser and subsequent detection of cry2±RFP using a FVX-BA585IF®lter.Single¯uorophore imaging allowed adjustments of the threshold settings and background subtractions to reduce the levels of crosstalk between the channels. To measure FRETsignals by acceptor photobleaching,a pre-bleached phyB±GFP image was acquired,and the¯uorescent intensity in a region of interest of4m m in diameter was recorded using the software provided by the manufacturer(Fluoview).Cry2±RFP¯uor-escence was then photodestroyed by repeated scanning with the568-nm laser line.A second post-bleach image of phyB±GFP in the same region of interest was acquired.After corrections for changes in image registration20,the¯uorescent intensities of the two phyB±GFP images(pre-and post-bleached)were compared.PhyB±GFP speckles that did not colocalize with cry2±RFP and colocalization of non-fused GFP and RFP were similarly processed.Received14June;accepted29August2000.1.Kendric,R.E.&Kronenberg,G.H.M.Photomorphogenesis in Plants(Kluwer Academic,Dordrecht,1994).2.Quail,P.H.et al.Phytochromes:photosensory perception and signal transduction.Science268,675±680(1995).3.Cashmore,A.R.,Jarillo,J.A.,Wu,Y.&Liu,D.Cryptochromes:Blue light receptors for plants andanimals.Science284,760±765(1999).4.Chory,J.Light modulation of vegetative development.Plant Cell9,1225±1234(1997).5.Briggs,W.R.&Huala,E.Blue-light photoreceptors in higher plants.Annu.Rev.Cell Dev.Biol.15,33±62(1999).6.Casal,J.J.Phytochromes,cryptochromes,phototropin:photoreceptor interactions in plants.Photochem.Photobiol.71,1±11(2000).7.Somers,D.E.,Devlin,P.F.&Kay,S.A.Phytochromes and cryptochromes in the entrainment of theArabidopsis circadian clock.Science282,1488±1490(1998).lar,A.J.,Straume,M.,Chory,J.,Chua,N.H.&Kay,S.A.The regulation of circadian period byphototransduction pathways in Arabidopsis.Science267,1163±1166(1995).9.Whitelam,G.C.&Devlin,P.F.Roles of different phytochromes in Arabidopsis photomorphogenesis.Plant Cell Environ.20,752±758(1998).10.Guo,H.,Yang,H.,Mockler,T.C.&Lin,C.Regulation of¯owering time by Arabidopsis photo-receptors.Science279,1360±1363(1998).11.Mockler,T.C.,Guo,H.,Y ang,H.,Duong,H.&Lin,C.Antagonistic actions of Arabidopsis crypto-chromes and phytochrome B in the regulation of¯oral induction.Development126,2073±2082(1999).12.Reed,J.W.,Nagpal,P.,Poole,D.S.,Furuya,M.&Chory,J.Mutations in the gene for the red/far redlight receptor phytochrome B alter cell elongation and physiological responses throughoutArabidopsis development.Plant Cell5,147±157(1993).13.Lin,C.et al.Enhancement of blue-light sensitivity of Arabidopsis seedlings by blue light receptorcryptochrome2.Proc.Natl A95,2686±2690(1998).14.Guo,H.,Duong,H.,Ma,N.&Lin,C.The Arabidopsis blue light receptor cryptochrome2is a nuclearprotein regulated by a blue light-dependent post-transcriptional mechanism.Plant J.19,279±287 (1999).15.Kleiner,O.,Kircher,S.,Harter,K.&Batschauer,A.Nuclear localization of the Arabidopsis blue lightreceptor cryptochrome2.Plant J.19,289±296(1999).16.Yamaguchi,R.,Nakamura,M.,Mochizuki,N.,Kay,S.A.&Nagatani,A.Light-dependent trans-location of a phytochrome B-GFP fusion protein to the nucleus in transgenic Arabidopsis.J.Cell Biol.145,437±445(1999).17.Kircher,S.et al.Light quality-dependent nuclear import of the plant photoreceptors phytochrome Aand B.Plant Cell11,1445±1456(1999).18.Wu,P.&Brand,L.Resonance energy transfer:methods and applications.Anal.Biochem.218,1±13(1994).19.Gadella,T.W.J.,van der Krogt,G.N.M.&Bisseling,T.GFP-based FRET microscopy in living plantcells.Trends Plant Sci.4,287±291(1999).20.Bastiaens,P.I.H.&Jovin,T.M.in Cell Biology:A Laboratory Handbook(ed.Celis,J.E.)136±146(Academic,New York,1998).21.Yeh,K.C.&Lagarias,J.C.Eukaryotic phytochromes:light-regulated serine/threonine protein kinaseswith histidine kinase ancestry.Proc.Natl A95,13976±13981(1998).22.Ni,M.,Tepperman,J.M.&Quail,P.H.PIF3,a phytochrome-interacting factor necessary for normalphotoinduced signal transduction,is a novel basic helix±loop±helix protein.Cell95,657±667(1998).23.Fankhauser,C.et al.PKS1,a substrate phosphorylated by phytochrome that modulates light signalingin Arabidopsis.Science284,1539±1541(1999).24.Choi,G.et al.Phytochrome signalling is mediated through nucleoside diphosphate kinase2.Nature401,610±613(1999).25.Ahmad,M.,Jarrillo,J.A.,Smirnova,O.&Cashmore,A.R.The CRY1blue light photoreceptor ofArabidopsis interacts with phytochrome in vitro.Mol.Cell1,939±948(1998)mond,A.I.&Earnshaw,W.C.Structure and function in the nucleus.Science280,547±553(1998).27.Somers,D.E.,Schultz,T.F.,Milnamow,M.&Kay,S.A.ZEITLUPE encodes a novel clock-associatedPAS protein from Arabidopsis.Cell101,319±329(2000).28.Nelson,D.C.,Lasswell,J.,Rogg,L.E.,Cohen,M.A.&Bartel,B.FKF1,a clock-controlled gene thatregulates the transition to¯owering in Arabidopsis.Cell101,331±340(2000).29.Carrington,J.C.,Freed,D.D.&Leinicke,A.J.Bipartite signal sequence mediates nucleartranslocation of the plant potyviral NIa protein.Plant Cell3,953±962(1991).30.MaÂs,P.&Beachy,R.N.Replication of tobacco mosaic virus on endoplasmic reticulum and role of thecytoskeleton and virus movement protein in intracellular distribution of viral RNA.J.Cell Biol.147, 945±958(1999).AcknowledgementsWe thank S.L.Harmer,T.F.Schultz and M.J.Yanovsky for critical comments of the manuscript.We are grateful to C.Lin for providing cry2cDNA,the cry2overexpression line and cry2antibody.We also thank A.Nagatani for the phyB±GFP construct and anti-phyB antibody.We thank G.Patterson and D.Piston for GFP spectra data,and Clontech for DsRFP spectra data.We are grateful to lar and T.K.Nomanbhoy for helpful discussion on FRETanalysis.Research support came from the NIH.P.M.was supported by a Novartis Agricultural Discovery Institute and P.F.D.was supported by an European Molecular Biology Organization long-term fellowship and the NSF.S.P.was supported by TSRI graduate program.Correspondence and requests for materials should be addressed to S.A.K.(e-mail:stevek@). ................................................................. Intracellular action of the cytokine MIF to modulate AP-1activity and the cell cycle through Jab1Robert Kleemann*,Angelika Hausser²,Georg Geiger*,Ralf Mischke*, Anke Burger-Kentischer*,Oliver Flieger*,Franz-Josef Johannes³, Thierry Roger§,Thierry Calandra§,Aphrodite Kapurniotu k,Matthias Grell²¶,Doris Finkelmeier*,Herwig Brunner*&JuÈrgen Bernhagen**Laboratory of Biochemistry/Institute for Interfacial Engineering and²Institute for Cell Biology and Immunology,University of Stuttgart,Nobelstrasse12,D-70569Stuttgart,Germany³Fraunhofer IGB,Nobelstrasse12,D-70569Stuttgart,Germany§Division of Infectious Diseases,Centre Hospitalier Universitaire Vaudois, CH-1011Lausanne,Switzerlandk Physiological-chemical Institute,University of TuÈbingen,Hoppe-Seyler-Strasse 4,D-72076TuÈbingen,Germany .............................................................................................................................................. Cytokines are multifunctional mediators that classically modulate immune activity by receptor-mediated pathways.Macrophage migration inhibitory factor(MIF)is a cytokine that has a critical role in several in¯ammatory conditions1±3but that also has endocrine4,5and enzymatic functions6,7.The molecular targets of MIF action have so far remained unclear.Here we show that MIF speci®cally interacts with an intracellular protein,Jab1,which is a coactivator of AP-1transcription8,9that also promotes degrada-tion of the cyclin-dependent kinase inhibitor p27Kip1(ref.10). MIF colocalizes with Jab1in the cytosol,and both endogenous and exogenously added MIF following endocytosis bind Jab1.MIF inhibits Jab1-and stimulus-enhanced AP-1activity,but does not interfere with the induction of the transcription factor NF k B. Jab1activates c-Jun amino-terminal kinase(JNK)activity and enhances endogenous phospho-c-Jun levels,and MIF inhibits these effects.MIF also antagonizes Jab1-dependent cell-cycle regulation by increasing p27Kip1expression through stabilization of p27Kip1protein.Consequently,Jab1-mediated rescue of®bro-blasts from growth arrest is blocked by MIF.Amino acids50±65¶Present address:Merck KGaA,Preclinical Research Oncology,D-64271Darmstadt,Germany.。
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IWALT 2000 Panel – 2Telemedicine and Tele - EducationRemote Tele-Education as an Alternative to Telemedicine Aboard the Aircraft Carrier USSGEORGE WASHINGTONPanel Chair: Les Folio, D.O., M.P.H., M.Sc., M.A.S.; William Sneeder, M.D., M.P.H. Residents in Aerospace Medicine, USAF School of Aerospace Medicine, Brooks Air Force Base, TXlesfolio@AbstractPurpose: A method of Basic tele-education as an alternative to telemedicine on the aircraft carrier USS GEORGE WASHINGTON is described. Aircraft carriers are often deployed for months, with limited medical specialty services. This method was developed to increase access to images by specialists at the medical center where patients would be seen once at port. Methods: Radiologic images were imported into Word and PowerPoint with simple windows keyboard commands. Images were then e-mailed to various specialists for consultation. Results: The quality of the attachments for diagnosis was satisfactory to the consultants and replies were received within 24 hours. There was no apparent difference in image quality between Word and PowerPoint and both used comparable memory space. Conclusion: Telemedicine, teleconsultation and tele-education may be successfully implemented in remote environments such as carrier operations at sea, using PowerPoint with attachments.1. IntroductionThe USS GEORGE WASHINGTON (figure 1) is a Nimitz class nuclear powered aircraft carrier, capable of DICOM compatible computed digital radiography and teleradiology. Tele-education was employed using digital radiology and commercial satellite Internet access during a deployment with training competitive exercises.FIGURE 1. Nimitz class aircraft carrier USS GEORGE WASHINGTON at port. Telemedicine, teleconsultation and tele-education, herein collectively referred to as telemedicine are well-established applications of computer and communication interactive technology, especially in remote locations. Medical groups on aircraft carriers represent a unique form of remote medicine, with the technology able to overcome obstacles of the usual lack of specialties and expertise.Medical capabilities on the Nimitz class carrier USS GEORGE WASHINGTON include: Digital radiology, Ultrasound, acute care, ER, ICU, OR, aviation medicine, environmental health and up to 60 inpatient beds with flexible expansion capability in case of combat or other mass casualty situation. Full medical personnel compliment includes: two flight surgeons, a General Medical Officer, a Physicians Assistant, a General Surgeon, Anesthesiologist, nurse, psychologist, and approximately 46 enlisted corpsman. The USS GEORGE WASHINGTON was the prototype carrier to utilize digital radiography with successful application of basic Picture Archiving System (PACS) on a departmental Local Area Network (LAN). [1]2. Materials / MethodsRadiographs were viewed with a 21 inch Hitachi CRT 2000 x 2000 pixel resolution in an exam room setting with dimmed overhead fluorescent lighting aboard the GW. Various radiographs were copied by right mouse click on the desired DICOM raw image (without the patient data) for comparison. They were then pasted into Word and PowerPoint with the paste command (control-v shortcut). Several images were compared in vertically tiled windows to compare diagnostic quality differences. Each case was reviewed in combinations of both Power Point and Word, to evaluate loss of diagnostic ability compared to the DICOM raw images.The entire radiographic image was imported from the clipboard onto the first PowerPoint slide and detailed enlargements were made on subsequent slides. Appropriate sizing was used to fit the imported images using the mouse. Images imported into Power-Point werethen e-mailed to various specialists for consultation, with satisfaction on the receiving end with timely replies and advice.FIGURE 2. Radiograph file size comparison of DICOM raw data images verses images in PowerPoint and Word. The quality degraded slightly converting from the .img raw data file to the .jpg. However, unlike .img, .jpg files are fully compatible with computers unable to display DICOM images. DICOM Radiographic images are periodically archived onto economical read-write CD, while the current patients (50-100) x-rays were readily accessible on the 30-gigabyte hard drive. Archiving DICOM onto CD Write at one dollar per CD is economical and convenient for placing into a patient folder or filing teaching cases. This readily available technology is markedly improved over previous optical disc methods. [2] The larger hard allow several weeks of use without the need to archive.Digital radiological images and clinical photos were saved as PowerPoint jpeg files and attached to email sent via satellite to CONUS-based specialists (such as, the orthopedics department) at the Naval Medical Center Portsmouth, which is the GW’s referral hospital when in-port at Norfolk, VA. The digital photos and radiographs in this limited analysis were sent as educational cases without any patient name, identification number or any other personal identifiable information. Follow-up emails and phone calls were made to discuss the patients’ history and other data that had been sent in separate messages. 3. ResultsAlthough there was no appreciable difference in quality, Power Point allowed for more natural importing of images including sizing, manipulating and printing. This was also true on the receiving end of the images on shore-based medical facilities. These images had barely discernable differences on the CRT monitor. Images included studies such as a chest x-ray PA and Lateral, Tibia-Fibula, and a 4th metacarpal. Although possible, laser printout quality provided only fair reproduction of subtle radiological details. Diagnostic quality and detail did not seem significantly degraded on the full image DICOM versus PowerPoint; especially with select magnified images. All abnormalities were seen well on images that were sent by email as attachments, and seen on the receiving end to satisfactory detail. Although this is subjective, the concept was demonstrated and seemed acceptable based on consulted specialists feedback for the pathology presented. Responses by specialists were returned within 24 hours of the original email consult request, with advice on diagnostic modalities and treatment recommendations.4. DiscussionTelemedicine is an electronic way to provide a medical skill set (e.g. radiographic interpretation, etc) to an area (aircraft carrier) where that skill set would not otherwise be available. Alternative ways to provide the same service include: (1) put a radiologist at sea; (2) super-train onsite PCM's (Primary Care Managers) in radiographic interpretation. These alternatives are less preferable to telemedicine because they are not cost effective.Tele-education about a current case by consulting a specialist allows for learning to take place while benefiting the patient. Using images sanitized for patient information avoids the legalities of patient data and confidentiality issues and streamlines the patient care. The tele-education approach also allows the specialist to “triage” and schedule the higher priority patients based on radiographs they have actually seen. [3] Other considerations in methods of transmission were bandwidth availability, potential loss of satellite signal during long transmission times required for larger digital radiology files, and consideration of obvious priority reduction of finite communication network time and access available to medical services during contingency deployments and actual combat operations. There should be traditional backup systems in case bandwidth is unavailable especially since bandwidth is at a premium during operational maneuvers/war. To assure that the onsite PCM's do not become technology dependent on telemedicine they should continue to receive usual recurrent medical skills training.Internet access was limited and inconsistent on this deployment, so writing teaching cases to a web site was not performed. Instead, email attachments were used to send photos and radiographs, with replies sent from the referred specialists by replying to the email transmissions. One important limitation to this proof-of-concept study design, was a lack of quantifiable resolution differences in the .jpg and .img image formats. Further research is recommended in this area.5. ConclusionsThis exploratory research attempted to demonstrate the feasibility of an alternative to DICOM technology that is more widely available, with minimum diagnostic quality loss. It was found that email attachments of Lossy compressed radiographs imported into PowerPoint was the most efficient and universally accepted method of sending radiographs to specialists without DICOM capability during our trial period at sea. The authors conclude that the more widespread and faster availability of the images outweighed the minimal loss in resolution, however, more research is needed in resolution comparisons. Review of several DICIOM verses PowerPoint images failed to reveal significant loss of diagnostic quality in that the pathology of interest was evident on the .jpg images sent and received. Getting the knowledge to the provider in a timely manner through tele-eduation allowed for more definitive patient care while establishing a case dialog with the specialist that would ultimately be seeing the patient once ashore.Turn around time with email attachments to specialist with replys within 24 hours’ verses multiple day turn around with centers with DICOM compatibility. More importantly, the images were electronically delivered to the specialists that were going to be seeing the patients once reaching port. Usual security and confidentiality issues were avoided by performing the tele-consultation as tele-education. Providing the images in PowerPoint allow specialists to easily incorporate interesting cases into their presentations. Perhaps this could serve as additional incentives for specialists to continually evaluate interesting cases in remote areas such as deployed aircraft carriers and hospital ships.Our sailors and troops injured or ill at sea cannot afford to wait for medical services as facility technology catches up. They can get it now with just-in-time technology. Our goal was to develop an acceptable system interface between the more advanced radiology system available shipboard and translate it into a readable, compact imagery data set that could be received and evaluated in a timely manner using systems now commonly available to specialty consultants, at the same hospital where the patients would physically go when ashore.6. AcknowledgementsThe authors greatly appreciate the cooperation and support from all the members of the Medical Group on the USS GEORGE WASHINGTON. Special thanks to Dr. Solomon, the ship’s Chief Primary care physician and Dr. Rocereto, the ship’s Senior Medical Officer for their highly professional assistance with this project.7. DisclaimerThe opinions expressed are strictly those of the authors, and do not necessarily reflect the opinions or policy of the Department of Defense, the US Air Force or the US Navy.8. References[1] Folio, L., Sneeder, W., Solomon, C. Rocereto, P., Applied Telemedicine, Teleconsultation And Tele-Education Aboard Uss George Washington Aircraft Carrier; Aviation, Space and Environmental Medicine (Abstract), March 2000, pp. 165.[2] Folio, L ., Stokes, R.,Frankfurter, J.“From Film to Filmless: Military Experience with Teleradiology in Korea;” Applied Radiology, July 95.[3] Karpf, B., “Navy Telemedicine Sails ‘Off-The-Shelf,” U.S. Medcine, February 2000, pp. 2, 28.[4] Buntic, R.F., Siko, P.P., Buncke, G.M., et.al., “Using the Internet for Rapid Exchange of Photographs and X-Ray Images to Evaluate Potential Extremity Replantation Candidates,” The Journal of Trauma, Injury, Infection and Critical Care, Vol 43, No.2, August 97, pp. 342-344.Complementary papers in this session relating to tele-education technologies:1. Banks: “Continuing medical education via IT: a viable alternative for female rural and remote general practitioners, disenfranchised from travel based options by family responsibilities”2. Arnott, Davis: "Recording the Use of Computer Aided Learning in Veterinary Anatomy: Results for a Complete Year"3. Togno: “Policy on Using Information Technology to improve rural health care”。