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Assume,. for. example,. that. the. encoder. generates. 600. pulses. per. revolution,. and. it. takes. 000. motor. revolutions. to. move. the. platform. from. one. position. to. another,. moving. the. platform. from. position. . to. position. 6. (5. positions). takes. 5000. motor. revolutions. or. 30,000. encoder. pulses.. In. most. practical. applications,. the. frequency. of. these. pulses. is. too. high. for. them. to. be. counted. with. inputs. that. are. not. associated. with. a. highspeed. counter.. Interrupts. S7-200. PLCs. incorporate. instructions. for. use. with. interrupts.. Interrupts. are. used. to. initiate. a. specific,. hort. PLC. program. segment,. called. an. interrupt. routine,. when. an. internal. or. external. event. occurs.. After. the. interrupt. routine. has. been. executed,. control. is. returned. to. the. main. program. Three. types. of. interrupts. are. supported. by. S7-200. PLCs,. communication port interrupts,. I/O interrupts,. and. timebased interrupts .. Communication. port. interrupts. are. used. to. control. a. communication. port. operated. in. Freeport. mode.. I/O. interrupts. are. used. to. respond. quickly. to. high-speed. I/O. transitions,. such. as. those. associated. with. high-speed. counters. or. pulse. train. outputs..Time-based. interrupts. allow. the. user. program. to. execute. an. interrupt. routine. on. a. cyclic. basis. Each. of. these. types. of. interrupts. has. an. associated. priority. that. determines. which. interrupt. is. processed. first. in. the. event. that. two. or. more. interrupts. are. requested. at. the. same. time.. Communication. port. interrupts. have. the. highest. priority. and. time-based. interrupts. have. the. lowest. priority. Pulse Training Output (PTO). S7-200. PLCs. have. two. PTO/PWM generators. that. create. either. a. high-speed. pulse. train. or. a. pulse. width. modulated. waveform.. One. generator. is. assigned. o. output. point. Q0. 0. and. the. other. to. output. point. Q0. .. When. a. generator. is. activated,. it. controls. its. respective. output. Pulse Train Output (PTO). is. used. to. provide. a. series. of. pulses. to. an. output. device,. such. as. a. stepper. motor. driver.. The. PTO. provides. a. square. wave. output. for. a. specified. number. of. pulses. and. a. specified. cycle. time.. The. number. of. pulses. can. be. from. . to. 4,294,967 ,295. pulses.. The. Pulse. Train. Output. has. a. 50%. duty. cycle.. This. means. the. pulse. is. off. for. the. same. amount. of. time. that. it. is. on. 63 The. number. of. pulses. and. he. cycle. time. can. be. changed. with. an. interrupt.. In. the. accompanying. example,. each. pulse. is. initially. on. for. 500. ms. and. off. for. 500. ms.. After. four. pulses,. an. interrupt. occurs. which. changes. the. cycle. time. to. 2. seconds,. . second. on. and. . second. off. . 1 sec 1 sec 500 ms Interrupt Occurs Pulse Width Modulation. (PWM) The. Pulse Width Modulation (PWM) function. provides. a. fixed. cycle. time. with. a. variable. duty. cycle.. When. the. pulse. width. is. equal. to. the. cycle. time,. the. duty. cycle. is. 00%. and. the. output. is. turned. on. continuously.. In. the. following. example,. he. output. initially. has. a. 0%. duty. cycle. (on. 0%,. off. 90%).. After. an. interrupt,. the. output. switches. to. a. 50%. duty. cycle. (on. 50%,. off. 50%). On Off On Off 10% Duty Cycle 50% Duty Cycle Interrupt Occurs The. PWM. function. can. be. used. to. provide. a. programmable. or. adjustable. control. of. machine. timing.. This. allows. machine. operation. to. be. varied. to. compensate. for. product. variations. or. mechanical. wear. And Much More. The. instructions. listed. in. this. section. are. only. examples. of. the. types. of. instructions. available. for. S7-200. PLCs.. The. full. instruction. set. includes. a. uch. broader. range. of. capabilities.. Refer. to. the. S7-200. System. Manual. for. additional. information. 64 Specialized Expansion Modules In. addition. to. the. expansion. modules. previously. discussed. that. provide. additional. discrete. or. analog. I/O,. several. expansion. modules. are. available. to. provide. communication. interfaces. or. specialized. I/O. functions. EM 241 Modem Module One. of. these. modules. is. the EM 241 Modem module.. This. module. supports. communication. between. a. computer. with. STEP. 7. Micro/WIN. and. an. S7-200. PLC. SF/DIAG EM 241 MODEM 241-1AA22-0XA0 S7-200 PLC with EM 241 Modem ModuleModem Computer The. EM. 24. provides. an. international. telephone. line. interface. and. supports. sending. numeric. and. text. paging. messages,. as. well. as. SMS. (Short. Message. Service). messages. to. cellular. phones.. This. is. useful. for. remote. diagnostics. and. maintenance,. machine. control,. alarm. systems,. and. general. communication. functions. In. addition. to. CPU-to-CPU. communication. via. a. telephone. line,. the. EM. 24. also. supports. Modbus. RTU. protocol,. a. protocol . that. has. been. widely. used. for. many. years. SINAUT MD 720-3 GSM/GPRS Modem Module SINAUT Telecontrol (Siemens Network Automation). ermits networking. of. individual. controls. and. control. systems. over. a. WAN. (Wide. Area. Network).. One. approach. for. providing. this. capability. is. SINAUT Micro.. This. is. a. simple. and. flexible. way. to. link. stationary. or. mobile. stations. to. a. master. control. center.. SINAUT. Micro. is. appropriate. where. smaller. amounts. of. data. have. to. be. transmitted. to. permit. monitoring. and. control. of. remote. stations. using. wireless. techniques. with. the. General. Packet. Radio. Service. (GPRS). of. the. Global. System. for. Mobile. Communication. (GSM). mobile. radio. network.. 65 The.SINAUT MD720-3 GSM/GPRS Modem module. and. associated. ANT794-4MR antenna. are. the. hardware. elements. used. to. connect. an. S7-200. PLC. into. a. SINAUT. Micro. system.. SINAUT Micro SC software. is. also. required. WinCC flexiible, Win CC SET S O C RS232 X1 SINAUT MD720-3 720-3AA00 SINAUT MD 720-3 GSM/GPRS Modem Module Antenna CP 243-1, CP 243-1 IT Communication Processors. Industrial. Ethernet. provides. a. proven. means. of. networking computers. and. a. variety. of. intelligent. devices.. CP 243-1 and CP 243-1 IT communication processors. are. used. to. connect. an. S7-200. PLC. to. an. Industrial Ethernet network. CP. 43-. and. CP. 243-. IT. communication. processors. can. be. used. to. connect. an. S7-200. PLC. via. Industrial. Ethernet. to. a. computer. running. STEP. 7. Micro/WIN.. This. allows. the. S7-200. PLC. to. be. configured,. programmed,. and. diagnosed. remotely.. In. addition,. an. S7-200. PLC. connected. to. an. Industrial. Ethernet. network. can. communicate. with. S7-200,. S7-300,. and. S7-400. PLCs. and. a. variety. of. other. devices. The. IT. functions. of. the. CP. 243-. IT. Internet. module. simplify. the. process. of. setting. up. a. control. system. that. can. email. diagnostic. infor mation. or. transfer. files. using.Internet. protocols. S7-200 PLC with CP 243-1 or CP 243-1 IT Communication Processor SF/DIAG SIMATIC S7-300 PLC CP 243-1 Ethernet CP 243-1EX00-0XE0 Industrial Ethernet Programming Device (PG) or Computer SIMATIC S7-400 PLC 66 EM 277 PROFIBUS-DP. Module. PROFIBUS DP. is. an. open,. international. fieldbus. communication standard. that. allows. a. broad. range. of. intelligent. devices. from. various. manufacturers. to. communicate. rapidly. and. efficiently.. This. reduces. wiring. costs. as. well. as. start-up. and. maintenance. expenses.. EM 277 PROFIBUS-DP module. allows. connection. of. the. S7-200. CPU. (CPU. 222. and. above). o. a. PROFIBUS-DP. network. as. a. slave.. Non-Siemens Controllers Other Intelligent Devices and Systems S7-200 PLC with EM 277 PROFIBUS DP Module SIMATIC S7 – 200 SF/DIAG RUN STOP 0 Other SIMATIC Controllers 2 4 0 2 8 X10 6 8 00 . 0 . 1 . 2 . 3 . 4 . 5 . 6 . 7 01 . 0 . 1 CPU 224 AC/DC/RLY X1 6 4 EM 277 PROFIBUS-DP CPU FAULT POWER DP ERROR DX MODE I0 .0 . 1 . 2 . 3 . 4 . 5 . 6 . 7 I1 .0 . 1 . 2 . 3 . 4 . 5 PORT 0 PROFIBUS DP I/O Systems Display Systems Computers CP 243-2 AS-Interface Master Module Actuator Sensor Interface (AS-Interface or AS-i). is. a. system for. networking. field. devices. such. as. sensors. and. actuators. ith. control. and. operator. interface. devices.. AS-i. replaces. the. extensive. parallel. wiring. often. used. to. connect. sensors. and. actuators. to. controllers. with. a. simple. 2-core. cable.. The. cable. is. designed. so. that. devices. can. only. be. connected. correctly. CP 243-2 AS-Interface Master module. allows. connection. of. the. S7-200. CPU. (CPU. 222. and. above). to. a. AS-I. network. as. a. master.. S7-200 PLC with CP 243-2 AS-Interface Master Module SIMATIC S7 – 200 SF/DIAG RUN STOP 00 . 0 . 1 . 2 . 3 . 4 . 5 . 6 . 7 01 . 0 . 1 CPU 224 AC/DC/RLY CM CER PWR APF SF B SET AS-Interface Master CP 243-2 AUP 0 1 2 3 5 6 7 8 9 10 11 12 13 14 15 16 1 7 18 19 20 21 22 23 24 25 26 27 28 29 30 31 31 I0 .0 . 1 . 2 . 3 . 4 . 5 . 6 . 7 I1 .0 . 1 . 2 . 3 . 4 . 5 DISPLAY 6GK7 243-2AX01-0XA0 AS-Interface Power Supply Slaves Repeater AS-Interface Power Supply Slaves PORT 0 100 meters S7-200 PLC with CP 243-2 AS-Interface Master Module SIMATIC S7 – 200 SF/DIAG RUN STOP 00 . 0 . 1 . 2 . 3 . 4 . 5 . 6 . 7 01 . 0 . 1 100 meters CPU 224 AC/DC/RLY CM CER PWR APF SF B SET AS-Interface Master CP 243-2 AUP 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 31 AS-Interface Power Supply Slaves Slaves AS-Interface Extension PlugI0 .0 . 1 . 2 . 3 . 4 . 5 . 6 . 7 I1 .0 . 1 . 2 . 3 . 4 . 5 DISPLAY 6GK7 243-2AX01-0XA0 PORT 0 200 meters 67 EM 253 Position Module. Position. control. describes. a. range. of. applications. that. involve. movement. with. varying. degrees. of. precision.. The. EM 253 Position module. is. a. simple. but. powerful. positioning. module. that. enables. the. user. to. control. position. sys tems. from. microsteppers. to. intelligent. servo. drives. (with. integrated. closed-loop. control). SF/DIAG MF MG P0 DIS P1 CLR PWR EM 253 Position STP ZP LMT RPS – + 253-1AA22-0XA0 S7-200 PLC with EM 253 Position Module EM 253 Features. Features. f. the. module. include: †¢. †¢. †¢. †¢. †¢. †¢. †¢. †¢. †¢. Provides. high-speed. control. with. a. range. from. 20. to.. 200,000. pulse. per. second Supports. both. S. curve. or. linear. acceleration. and. deceleration Provides. a. configurable. measuring. system. that. allows. you. to. enter. data. as. engineering. units. (such. as. inches. or. centimeters). or. as. a. number. of. pulses Provides. configurable. backlash. compensation Supports. absolute,. relative,. and. manual. methods. of. position. control Provides. continuous. operation Provides. up. to. 25. motion. profiles. with. up. to. 4. speed. changes. per. profile Provides. our. different. reference-point. seek. modes. with. a. choice. of. the. starting. seek. direction. and. final. approach. direction. for. each. sequence Provides. removable. field. wiring. connectors. for. easy. installation. and. removal 68 Expansion Modules for Temperature Measurement Two. S7-200. PLC. expansion. modules. are. available. for. accurate temperature. measurement, EM 231 Thermocouple module and EM 231 RTD module. EM. 23. Thermocouple. module. provides. analog. inputs. for. thermocouples.. A. thermocouple. is. a. temperature. sensor. made. from. two. dissimilar. metals. joined. at. a. point. called. a. junction..A. thermocouple. produces. a. small. voltage. that. is. dependent. upon. temperature.. Various. types. of. thermocouples. are. available. for. use. in. different. temperature. ranges.. Two. versions. of. EM. 23. Thermocouple. modules. are. available,. one. for. four. thermocouples. and. one. for. eight. thermocouples.. Each. version. is. compatible. with. J,. K,. T,. E,. R,. S,. or. N. thermocouples,. but. the . thermocouples. used. with. a. specific. module. must. be. of. the. same. type. EM. 23. RTD. module. provides. analog. inputs. for. resistance temperature detectors (RTDs).. An. RTD. is. a. temperature. sensor. made. rom. a. metal,. such. as. platinum,. nickel,. or. copper,. that. varies. in. resistance. in. a. predictable. manner. as. temperature. varies.. Two. versions. of. the. EM. 23. RTD. module. are. available,. one. with. two. analog. inputs. and. one. with. four. analog. inputs.. Either. version. can. be. used. with. a. variety. of. RTD. types,. but. the. RTDs. used. with. a. specific. module. must. be. of. the. same. type. SIMATIC S7 – 200 SF/DIAG RUN STOP 00 . 0 . 1 . 2 . 3 . 4 . 5 . 6 . 7 01 . 0 . 1 CPU 224 AC/DC/RLY +24 VDC SF EM 231 AI4 – TC I0 .0 . 1 . 2 . 3 . 4 . 5 . 6 . 7 I1 .0 . 1 . 2 . 3 . 4 . 5 243-7PD22-0XA0 PORT 0S7-200 PLC with EM 231 Expansion Module A+ A- B+ B- C+ C- D+ D- A+ A- a+ a- B+ B- b+ b- M L+ Configuration M L+ Configuration + 24 VDC Refer to the S7-200 Programmable Controller System Manual for Configuration DIP Switch Positions + 24 VDC Refer to the S7-200 Programmable Controller System Manual for Configuration DIP Switch Positions EM 231 Thermocouple module, 4 Input Version EM 231 RTD module, 2 Input Version 69 SIWAREX MS Weighing Module SIWAREX MS Weighing module provides.. a. simple,. easy to. install. approach. for. weighing. and. force. measurement. applications.. SWAREX. MS. Weighing. module. is. designed. to. measure. the. oltage. produced. by. sensors. commonly. used. to. measure. weight,. force,. or. torque. SIWAREX. MS. is. easily. integrated. into. an. S7-200. PLC. system. as. an. expansion. module.. This. makes. information. obtained. from. SIWAREX. MS. available. to. other. components. of. the. automation. system.. In. addition,. Siemens. offers. a. wide. variety. of. compatible. sensors. and. other. components. S7-200 PLC with SIWAREX MS Module SIMATIC S7 – 200 SF/DIAG RUN STOP 00 . 0 . 1 . 2 . 3 . 4 . 5 . 6 . 7 01 .0 . 1 CPU 224XP DC/DC/DC SF I NET p —I SIWAREX MS ->00< >< T Tp T o SIWAREX R Load Cell Touch Panel TP 177micro 0 Review 6 .. Three. types. of. SIMATIC. counters. available. in. the. S7-200. instruction. set. are. ____________,. ____________. and. ____________. 2†¦ CPU. 22. and. CPU. 222. support. ____. high. speed. counters.. CPU. 224,. CPU. 224XP ,. CPU. 224XPsi,. and. CPU. 226. support. ____. high. speed. counters. 3.. S7-200. PLCs. have. two. ___________. that. create. either. a. high-speed. pulse. train. or. a. pulse-width. modulated. waveform. 4.. ________. and. ________. communication. processors. are. used. to. connect. an. S7-200. PLC. to. an. Industrial. Ethernet. network. 5.. _________. module. allows. connection. of. an.S7-200. CPU. (CPU222. and. above). to. a. PROFIBUS-DP. network. as. a. slave. 6.. _________. module. allows. connection. of. an. S7-200. CPU. (CPU222. and. above). to. an. AS-I. network. as. a. master. 7 .. Two. v ersions. of. EM. 23. Thermocouple. module. are. available,. one. for. ____. thermocouples. and. one. for. ____. thermocouples. 8.. Two. versions. of. EM. 23. RTD. module. are. available,. one. for. ____. RTDs. and. one. for. ____. RTDs. 7 Review Answers Review 1 Review 2. . Review 3. Review 4. ). a:. input. module,. b:. CPU,. c:. output. module,. d:. programming. device,. e:. operator. interface;. 2). 2;. 3). 6;. 4). 00,. 000. 0000,. A. ). discrete;. 2). discrete;. 3). CPU;. 4). Ladder. logic;. 5). Statement. list,. function. block. diagrams;. 6). scan;. 7)024;. 8). firmware; 9). RS-485. ). CPU. 22,. CPU. 222,. CPU. 224,. CPU. 224XP ,. CPU. 224XPsi,. CPU. 226;. 2). b;. 3). 2,. 7;. 4). 8,. 6;. 5). 4,. 0;. 6). Q0. 3;. 7). DIN. ). a:. box,. b:. normally. open. contact,. c:. coil;. 2). AND. Function. -. a:. 0,. b:. 0,. c:. 0,. d:. ,. OR. Function. -. e:. 0,. f:. ,. g:. ,. h:. ;. 3). I0. ,. I0. 0,. Q0. 0. ). 224XP;. 2). On-Delay. Timer. (TON),. Retentive. On-Delay. Ti mer. (TONR),. Off-Delay. Timer. (TOF). ;. 3). 3276. 7. seconds;. ). Retentive. On-Delay. Timer. (TONR);. 5). On-Delay. Timer. (TON),. Off-Delay. Timer. (TOF),. Pulse. Timer. (TP). ). Count. Up. Counter. (CTU),. Count. Down. Counter. (CTD),. Count. Up/Down. Counter. (CTUD);. 2). 4,. 6;. 3). PTO/PWM. generators; 4). CP. 243-,. CP. 243-. IT;. 5). EM. 277. PROFIBUS-DP 6). CP. 243-2. AS-Interface. Master;. 7). 4,. 8;. 8). 2,. 4.. Review 5. Review 6. 72 73 Final Exam You. can. test. your. knowledge. by. taking. the. final. exam. for. this. course. online. at. http://www. usa. siemens. com/step.. This. web. page. provides. links. to. a. variety. of. our. quickSTEP. online. courses.. To. complete. he. final. exam. for. this. course,. click. on. the. Basics of PLCs. link.. Next,. move. your. mouse. over. to. the. left. so. that. the. navigation. bar. pops. out. and. select. the. Final Exam. link.. The. final. exam. page. will. appear.. Before. taking. the. final. exam,. it. is. recommende d. that. you. delete. the. temporary. files. on. your. computer.. For. most. versions. of. Internet Explorer,. you. can. do. this. by. selecting. Internet Options. from. the. Tools. menu. and. then. clicking. on. the. Delete Files. button.. If. you. do. not. perform. this. step,. you. may. see. a. score. of. 0%. after. you. submit. your. xam. for. grading. After. you. complete. the. final. exam,. click. on. the. Grade the Exam. button. at. the. bottom. of. the. page.. Your. score. on. the. exam. will. be. displayed. along. with. the. questions. that. you. missed.. If. you. score. 70%. or. better. on. the. exam,. you. will. be. given. two. options. for. displaying. and. printing. a. certificate. of. completion.. The. Print Certificate. option. allows. you. to. display. and. print. the. certificate. without. saving. your. score. in. our. database. and. the. Save Score. option. allows. you. to. save. your. score. and. display. and. print. your. certificate. 74

Reformation of the 16th Century

The Protestant Reformation ignited a religious reform movement that separated the western Christian church into Catholic and Protestant groups. Martin Luther embarked on a Journey to start the religious reform movement; there were other developments before him that set a foundation for a religious alteration in the sixteenth century.The Protestant Reformation allowed for Protestantism to flourish throughout Europe, united the Roman Catholic Church with Christian denominations, enabled people to develop independent thinking and creative, fostered etermination in people to attain religious and political freedom, and allowed for Christianity to evolve permanently throughout history. Johannes Gutenberg of Mainz helped produce the development of printing from a moving type. In Europe there were thousands of printers that published religious books like the Bible, sermons, Latin and Greek classics, legal handbooks, and works on philosophy.This type of development manifested an â€Å"immedi ate impact on European intellectual life and thought† (Duiker 429). This kind of invention fostered creative thinking and determination in people to study scholarly resources. Printing allowed the European civilization to disperse new religious ideals. This communication throughout Europe played a major role in enabling people to acquire newfound knowledge and formulate beliefs of their own. Many historians state that Desiderius Erasmus â€Å"laid the egg that Luther hatched† (History 1). Erasmus was an influential Christian humanist.Christian humanism or northern Renaissance humanism combined the ideas of the classical Italian Renaissance with the ideas of early Christianity. According to Christian humanism, a society must alter the human beings that compose it. They strongly believed that people are smart enough to formulate their own ideals and beliefs. They did no have to solely depend on an irrational religion for happiness and salvation. Erasmus was most influenti al in teaching that religion should be a philosophy for a direction of life, not a source of arbitrary practices and rituals, which is what medieval religion emphasized.Erasmus also placed an emphasis on teaching the philosophy of Jesus Christ. Erasmus did not approve of the abuse of power of inside the church. The opposition towards a dogmatic religion encouraged the Christian and northern Renaissance humanists to stand against the orruption of the Catholic Church; they gave mankind hope and encouragement to believe in themselves, and to fight for freedom of religion. Another factor that influenced people to start a reform was the corruption of the Catholic Church. The all-powerful Roman Catholic Church labeled any non-believer as a heretic; punishment included being burned at stake.The Catholic Church's power was assembled over centuries, and depended on the lack of religious education and unawareness of people. People's primary motivation of believing in the Catholic Church came from the belief of salvation to heaven though the Catholic Church. Any other belief was disregarded. The realization that many popes were not committing to the needs of the church and people outraged many people in Europe. Many popes' interests were not focused on the spiritual uprising of religion, but worldly interests like power and wealth The process ot salvation was becoming a traud.This ignited hate and bitterness towards the Catholic Church and stimulated the idea for change and reform. The Catholic Church strongly emphasized that good works and strong faith were the gateway for personal salvation. Martin Luther had a strong opposition o that idea, even as a monk and professor at the University of Wittenberg. Through personal study and dedication to religion he discovered that no weak and powerless human being could do enough good works to achieve salvation. He believed that through a powerful faith and the sacrifice of Jesus Christ, that alone was enough for people to achiev e salvation. Justification by Faith† (PBS 1) became the primary belief of the Protestant Reformation. Luther's repulsion of Catholic Church teachings enabled him to create the Ninety-Five Theses, which opposed the sale of indulgences and revolted against clerical abuses. According to many historians this was the beginning of the Protestant Reformation. The Ninety-Five Theses were quickly created and disseminated throughout Germany. Luther encouraged German royalty to overthrow the idea of salvation though the Catholic Church.His idea of salvation through faith alone was widespread and becoming acceptable to many people. The Roman Catholic Church was outraged and did not accept his view of salvation. The Catholic Church believed that he was to be excommunicated from the church in 1512. When Luther was summoned to appear to the Holy Roman Empire and to â€Å"recant the heresies† (Duiker 431). He responded by rebuffing and making a statement that â€Å"became the battle c ry of the Reformation† (Duiker 431). This angered the members of Reichstag, who ordered Luther to be detained.Luckily for him, Fredrick of Saxony protected him. A revolution was manifested upon Luther's religious ideals; this revolution changed the European Civilization forever. Luther's popularity fostered from hatred and dissatisfaction with the greed of wealth power, corruption and dogmatic ideals of the Roman Catholic Church. The marriage between Luther and Katherina von Bora demolished the idea of clerical celibacy; his marriage ighlighted the life of a Protestant minister with a family. German rulers strongly supported Luther; they quickly acquired control and supervision of the German churches.Luther emphasized that the churches focus on the preaching the Word of God and Bible study. In Luther's eyes in was necessary to abolish Catholic Mass and gravitate towards new religious services. All of these changes highlight the metamorphosis that Christianity was experiencing. People who believed and promoted these new ideals were complacent with a new religious experience. In 1 519, Charles V ruled a vast empire, composed of Austrian lands, part of Italy, Spain, nd other territories. Though election he was granted the Holy Roman Empire title.His idea was to â€Å"preserve the unity of his empire in the Catholic faith†(Duiker 432). By this time, Germany detached it self from the idea of imperial ruling; this disintegrated any loyalty to the Holy Roman Empire. In 1546, Lutheranism was the prominent belief and practice throughout Germany. Charles Vs effort to bring combat and war to disintegrate Lutheranism was a failure. This failure encouraged Charles V to create the Peace of Augsburg, which granted freedom to German states to choose between Catholicism and Lutheranism.The expeditious spread of Lutheranism and Protestant beliefs reassured that Christian unity was no longer a possibility. Switzerland had a prominent role in the Protestant Reformati on. The revolution sparked upon Ulrich Zwingli. Zwingli ignited religious changes in member states of the federal states of Switzerland and the city of Zurich. Evangelical reforms took over these areas. For example, Mass was replaced by services that promoted biblical study and prayer, relics were destroyed, and pope's authority was no longer accepted.His primary source of support came from the people of Zurich and the agistrate, known as Mark Reust. Zwingli created an alliance with the German rulers, and Martin Luther. In 1513, rage and dissatisfaction festered between Catholic states and the Swiss Protestants. This created a war that led to Zwingli's death, his responsibility and loyalty to manifest change in Switzerland was handed down to John Calvin. John Calvin was a practical theologian who organized the Protestant reform. His conversion to Protestantism led him to escape his native France to Switzerland.His beliefs coincided very closely with Luther's beliefs. A unique belief that he promoted was the idea of predestination. Predestination was a belief that â€Å"God predestined some people to be saved, and the others to be damned, both who He would admit to salvation, and whom He would condemn to destruction† (PBS 1). Calvin also emphasized absolute sovereignty of God; he called it the â€Å"power, grace, and glory of God†(PBS 1). His reputation and credibility arose from his publication of the Institutes of the Christian Religion, an integration of Protestant beliefs and stands.In 1536, he expanded his revolutionary Protestant reform to the city of Geneva. Calvin created The Consistory, a governing body that attested to moral virtue nd discipline, and enabled a systematic way of including clergy and laymen in church service. The success that was fostered upon Geneva, allowed for missionaries to travel throughout Europe and infiltrate all of Europe with Protestantism. The city of Geneva stood as a Protestant citadel that gave success to th e religious reform. The Anabaptists were radical promoters of the Protestant faith, who also played a pivotal role in the reformation.Their concern was to return to practice and religion of early Christianity, they emphasized equality in all believers. Separation of Church and State was something they strongly advocated. They did not see any benefit of the government dictating the Jurisdiction of religion. The Anabaptists opposed the idea of baptism in infants. Magisterial Protestants and Roman Catholics grew hostility and hatred towards the Anabaptists because of that belief. That is why they were heavily persecuted in the sixteenth century. The English Reformation played a pivotal part in the Protestant reformation.King Henry the VII was seen as selfish glutton. Surprisingly his strong desire to divorce Catherine of Aragon, his first wife made the English Reformation a more political than religious cause. The pope refused to grant Henry VII's request to divorce Catherine of Aragon for Anne Boleyn. His attention then focused on the archbishop of Canterbury, head of the highest court in England. The archbishop condoned a divorce and annulled King Henry VII's first marriage. Soon after Anne became queen she had a child, whom was a girl.This infuriated King Henry VII because he did not have a son to inherit his throne. This anger encouraged Henry VII to promote separation of the Church of England with Rome. This was known as the Act of Supremacy in 1534, it declared that the King was in complete control of the doctrine and Church of England. During Henry VII's reign, Archbishop Cranmer attempted to reform the administrative and religious ruling. Shortly after this, Henry VII died and left his ruling to his underage son. Cranmer took advantage of this situation by directing the Church ot England in a Protestant direction.T manifested new Protestant services and ideals in the Church of England. Mary a Catholic succeeded Edward VI. Her nickname â€Å"Bloody Mary' was no understatement. Her primary concern was for the Church of England to return to Catholicism; she murdered more than 300 heretics who opposed Catholicism. Ironically, her actions ed for England to become more Protestant than ever before. By the mid-sixteenth century, the Roman Catholic Church no longer dominated Europe. Protestantism was infiltrating all of Europe and becoming more dominating and accepted.Due to this nature, the Catholic Church underwent a reformation of its own known as the Catholic Reformation. The Catholic is divided in to three parts known as the Jesuits, a reformed papacy, and the Council of Trent. Ignatius of Loyola, Spanish noblemen, founded the Society of Jesus. This society accepted the directing of the pope and promoted the principals of obedience to the papacy, and hierarchical ruling of ociety and civilization. They fostered missionaries to explore Asian countries to convert people to Catholicism The reformed papacy was another aspect of the Catholi c Reformation.Pope Paul Ill took the initiative to create a reform to team to hold Catholic Church's accountable and enforce discipline. Through this he was able to create the Council of Trent and gave recognition to the Society of Jesus. In 1545, church officials gathered in the city of Trent started the Council of Trent. This Council of Trent met continuously though three different sessions in the sixteenth century. Their ideas coalesced and they came to the conclusion that the â€Å"selling of indulgences was prohibited, both faith and good works were required for salvation, and only the church could interpret scripture† (Duiker 438).This manifested one doctrine and religion under the Roman Catholic Church. The Protestant Reformation had significant impacts in the foundation of our world nations. This impact allowed for the re-establishment of political and religious views throughout the European civilization, challenged the authority of the Catholic popes, and led to mode rn concepts of democracy. This reformation is key to understanding how religion has laced an importance in the foundation and building of every nation in our world.The fight for equality and freedom will always be an obstacle throughout history; understanding the struggle, rebellion, and motivation to fght that many people faced in reformation, allows for our world to become educated and motivated to strive towards a better world.

Investigate if small villages can become suburbanised over time, and what factors will affect this

Aim: In this piece of coursework, my aim is to investigate if small villages can become suburbanised over time, and what factors will affect this. For this investigation, we have chosen to look at Pirbright, a small village just outside of surrey. Hypothesis: Is Pirbright a Suburbanised Village? To prove the hypothesis I will need to find out: * Where is Pirbright located? * What is it like? * How has it changed over the years? * What are the reasons? * What are the consequences of these changes for the long-term residents, the businesses and the new comers? Methods Of Research: In order to find answers to these questions and to prove the hypothesis I used various methods of research these will be: Questionnaire – We made up our own questionnaires in groups of four. The reason we thought this would be a good idea was so that we could get opinions from people who have been living in Pirbright and learn more about these people. We surveyed 10 different people and we tried to ask various types of people such as pensioners, young students and adults. In order to do this we asked at different times of the day. Walk To Investigate Services And Shops – We walked around Pirbright to investigate the different types of services they had to offer. Walk To Investigate Land Use And Location – We also walked to and around Pirbright to investigate the uses of land and the layout of the village. Environmental Quality Survey – This was a survey that had already been prepared for us. This helped us to decide whether or not the village was an attractive place to live. We completed the survey in different parts of the village. Research – I will find out any other information from resources such as the Internet. What is a Suburbanised Village? A Suburbanised village is a village that people from nearby towns and cities have moved into, gradually changing the character of the village and making it more urbanised. A suburbanised village change its functions from being a rural agricultural village to being a suburb of a town. Suburbanised villages can sometimes also be called commuter settlements or dormitory towns, this is because the village is used by residents who live and stay there to travel to work in nearby towns or cities. Since the 1980's people have been moving out of major cities such as London, -Reading and Guildford to get away from things such as crime, pollution and traffic congestion in cities. This is called counter urbanisation. The characteristics of a suburbanised village are; * Housing becomes more expensive leaving local people not being able to afford them. We would expect to find new houses built for newcomers, as well as old farmhouses that have been modernised for rich city commuters. * The need for more houses and extra services is required to cater to the needs of the newer residents * There are more newcomers than original residents; this can cause social problems within the village. * Cars, noise and litter cause more pollution than before. * The village is expanded to house more residents. * More cars cause congestion. * The village is forced to change to become more modern; this can also result in it becoming damaged. * People move to rural villages that are commuting distance from their place of work, so a suburbanised village must have access to railway and motorway links. * Newcomers will shop in cities and out of town shopping centres, so we would not expect to find medium or high order shops. People move into villages for many different reasons, these are; * People find towns and cities to be polluted, unattractive and very congested. * Wealthy commuters can afford to buy bigger and more attractive houses in villages and since transport links have improved these people can still easily commute to work in the city. * Retired people prefer to enjoy they leisure time in a peaceful and quieter environment. * People find it to be safer in villages. * There are newer houses being built, these sell for cheaper than they do in cities and towns. How can location affect the sub urbanisation of a village? The location of a village can be affected by its location. If a village were located near a large city, it would be easier to make use of the services provided in the city. This would be useful because there will be more services in the city such as hospitals, large shopping centres or a wider ranger of schools. If a village were located far away from a city it would be harder for the village residents because they would have to do with the only services provided in the village as it would be hard to get to the city. This would be inconvenient because usually there are not many services in a village. If a village were located near good roads that lead into the city it would be easier for people to get to the city, there probably would be a bus or some sort of public transport that would lead into the city. If a village is isolated with poor transport links it would make access in and out of the village difficult. This not only means that if would be for village residents to get to a city but also that visitors would find the village hard to get to. If a village is located on hilly land it would be difficult to build new buildings whereas if the village was built on flat land it would be easier to improve the village with new buildings and services. Where is Pirbright? The village that we are studying is called ‘Pirbright' it is located in Surrey, southeast England (see map below). Pirbright is located near Guildford, Bracknell, Woking, Farnborough and Camberley. The roads that connect these towns to Pirbright are A322, A3214, A320 and A323. These roads are good for commuters because they can travel to and from work in different towns quite easily. If they do not have cars then there are good rail links into other close by towns. There is no train station in Pirbright however the closest one is in Basingstoke. It takes approximately one hour to travel from Pirbright into central London. From the map below I can see that Pirbright very close to London and also to other cities, where good jobs are available. The land that Pirbright is built on is mainly countryside and there are many woodlands and open fields in and around the area. The land is mainly greenery and some areas are quite steep. The land is fertile, which makes it good for farming. There are also good communications in Pirbright. Map 1: Southeast England What is Pirbright like? Pirbright is a very attractive, yet small village. The population is approximately 3644 people, this has changed drastically over the past hundred years when there was only a few hundred people living in Pirbright. It has a large green in the centre of the village (see picture 1). The grass is always short and well kept. There are no signs of vandalism or litter. Around the green there are a few phone boxes and three bus stops, one going towards Woking and the other two going towards Guildford. The bus stops are connections to Guilford, Woking and other surrounding towns. There are a few shops around the green as well; these include a newsagent, a butcher, an antique shop and two pubs; The White Hart and The Royal Oak. There is a large pond and a children's playground on the green as well. Pirbright formally contained a post office, which was then forced to shut down because of robbery. This shows that the crime rate in Pirbright has increased. A mobile library visits Pirbright once a week. Picture 1: views of the green The church in Pirbright is called St Michael's church (see picture 2) and is around the green. There is only one school in Pirbright, Pirbright County Primary. It is only a primary school for five to eleven year olds. This means that when children are ready for high school they will have to travel to a nearby city or town. Most children travel to Guildford and Woking, as these are the closest and easiest to town to get to. Picture 2: St Michael's church Many attractive houses surround the green (see picture 3). These houses are mainly built before the 1940s. The houses are bigger in comparison to the sizes of housing in London, and are also much cheaper. An average three-bedroom house would cost approximately à ¯Ã‚ ¿Ã‚ ½385,000. The houses have extra garden space, bigger drive ways and more space to build extensions because the houses are spread out from each other. Picture 3: Houses around the green Below is a land-use map, to show the services available in Pirbright, and the ages of the houses. People in Pirbright We visited Pirbright on a typical mid-week afternoon to find out about the types of people that live there, we found out this information by asking people to fill out a questionnaire that we had designed. These are the results we came back with. I also did some extra research to find out facts and figures of Pirbright. People's opinions on Pirbright WHAT LEISURE ACTIVITIES ARE THERE IN THE AREA Tennis Karate Golf Cricket Bowles Dance WHAT DO YOU LIKE ABOUT PIRBRIGHT? Quiet Friendly People Plenty For Children To Do Peaceful Area Countryside Good Community WHAT DO YOU DISLIKE ABOUT PIRBRIGHT? Does Not Have a Post Office Is Not Sociable Services and Shops WHAT SERVICES ARE USED IN THE AREA WHO DO THEY CATER FOR? Newsagent Everyone in the village, most people use the newsagent everyday. Pubs Used mainly by adults, as a place to relax. Antique Shop Used by the wealthy residents, once only a few times a year. Butcher Used to provide food for all residents, used once a week. Mobile Library Used by anyone that wants it. Public Transport Used mainly by the retirees and students travelling to high school. Public Park Used by everyone in the village. Village Hall Used for meetings and clubs etc. dance clubs. Comparing Pirbright in 1871 to Pirbright today From the two maps I can see that Pirbright has drastically changed over the past 30 years. There was much more open space in 1871 but now a lot of that land has been used to build things such as houses and other services. The area that Pirbright occupies has expanded; more houses are being built so the village has had to expand to cater for the extra people. The land use has changed from mainly being farmland to having lots of houses and other buildings built on it. From the map of 1871 I can see that there were only a few houses whereas most of Pirbright is covered by houses now. More services have been added in Pirbright for example there used to be only one pub in 1871 but another one has been built now. Overall Pirbright has grown over the years and the open land has been used to build houses on. Conclusion After analysing the statistics and information about Pirbright I have come to a conclusion that all the evidence shows that Pirbright has become a suburbanised village. In 1915, we would have expected people to have jobs such as farmers, woodcutters, small village owners, ground keepers for private estates, housemaids and stable workers. The sort of people that would want to live there now are upper-class people as they do not usually have to work, retired people as they do not have to work either and people whom do not want to live in London, but still be based near London. The facts that point to the conclusion that Pirbright has become suburbanised are: * The population has greatly increased over time and is continuing to do so. Shown by the census figures. * The village has become larger. Many of the old houses are in the centre of the village and the newer ones have been built on the outskirts. This proves that new houses are being built for commuters and other residents wishing to move to Pirbright. This is shown in the land use map. * Most residents are middle ages and are high-skilled professionals – this proves that they are commuters as Pirbright does not offer and high-skilled professions. This is shown in the data that I collected on the field trip. * The home of most residents are detached or semi-detached. This proves that they are expensive and can only be afforded by wealthy commuters. * Most village residents own their own homes or are currently buying. This shows that the average Pirbright resident can afford houses; this is because mainly commuters live in Pirbright. * More then half of the residents own their own car. This also proves that a lot of commuters live in Pirbright, as they need a car to travel to work. * Most residents take their car to work, not public transport. * Most of the residents have lived in the village for 30 to 40 years. * The village is gradually becoming more modernised. The newcomers are transforming the village. * There are no high order shops, meaning residents have to visit a town or city for shopping centres. * Houses are becoming more expensive. The value is increasing as more people want to live in Pirbright and wealthy people can afford them. All these facts that I have stated link back to my theory on suburbanised villages, Pirbright has successfully gained nearly all the characteristics of a suburbanised village. Pirbright has gone from being a unknown small hamlet, to becoming a modern and attractive growing villages used mainly by commuters and retired people. I have found the main reason people move from large towns and cities into Pirbright is because it is a quiet and peaceful area. The long term residence will live closer to the village centre and the newer residence will be living on the edge as this will be where the new houses will be built. Evaluation I feel that this piece of coursework has greatly widened my knowledge of suburbanised villages. I did not thoroughly understand it at the beginning however after undertaking the research about the changes in Pirbright I have come to understand how and why small villages change their function to become urbanised. I feel I have done well in this coursework as I did a lot of research and used many new ICT skills that I have leant. I could improve the coursework by getting a broader range on data, I visited Pirbright on a working day therefore I did not get enough information about residents who may have been at work that day. To improve this now I would visit Pirbright on a weekend and carry out my research then. However overall I believe I have created a good report on how and why Pirbright has become suburbanised.

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Closing Guantanamo Prison - Essay Example Indeed, the United Kingdom has even called for the United States to shut down Guantanamo Bay (‘UK told US won’t shut Guantanamo’). This essay considers a number of contentions regarding the importance of keeping Guantanamo Prison a functioning detention center, and then ultimately argues that after considering the arguments and evidence that the detention facility should be closed. When considering the potential closing of Guantanamo Bay prison there are a number of objections individuals have levied to this proposal. One of the central objections in these regards concerns the intelligence that could have been potentially gained in the interrogation procedures that were conducted there during and immediately after the conflicts in Iraq and Afghanistan. During this period extensive water-boarding interrogation procedures were conducted with individuals that had been identified as major terrorists and believed to have information that was essential to United States war efforts in the Middle East. Governmental investigations into Guantanamo have even at times supported the detaining of prisoners in the facility. A recent Washington Post report stated, â€Å"A Justice Department-led task force has concluded that nearly 50 of the 196 detainees at the U.S. military prison at Guantanamo Bay, Cuba, should be held indefinitely without trial under the laws of war, according to Obama administration officials† (Finn 2010). Still, many reports have indicated the growing levels of torture implemented at the facility. Hyland (2004) indicated that, â€Å"Three Britons freed from Guantanamo Bay in March have released a 115-page dossier accusing the US of carrying out torture and sexual degradation at the military concentration camp in Cuba.† Even as in large part these activities were deemed near torture by human rights groups, they were still allowed to occur as they were carried out within the confines of Guantanamo. When considering the evid ence, it’s shown these interrogation methods are entirely ineffective. Research indicates that overly intensive interrogation methods have been demonstrated to be ineffective in gaining information, and oftentimes lead to false information (Butler 2007). This is particularly clear in the instance of Guantanamo Bay interrogations. Butler (2007, pg. 63) writes, Often all that is gained by aggressive interrogative techniques are false confessions. For example, Asif Iqbal gave interrogators a false confession. After extensive interrogation, he finally admitted that he was, indeed, in the videotape with Osama Bin Laden†¦Later, the British intelligence found proof that Asif Iqbal†¦had been in England at the time the video was made Indeed, the argument is that rather than actually being an effective method of gathering intelligence information, these interrogation methods resulted in false information being gathered. When one considers the issues more intently further evid ence points to this rebuttal being correct. Debeaux (2005) even indicated that a number of detainees were even gathered through a bounty system that was in place between Afghanis and United States’ forces; when one considers this, the increased potential for misinformation is greatly increased. Ultimately this makes it so that the main benefit of keeping this prison open is negated

How Technology Affects the Business Environment Essay Example for Free

How Technology Affects the Business Environment Essay During the last century, the world has been through a technological revolution. We have in few years moved from being a world where it could take months to receive a message, to being a world where it takes microseconds. Within most industries, technology is a very important factor to succeed. Especially in certain industries, like energy, transport and financial services, technological innovations have been vital to make those industries develop into what they are today. Companies are always struggling to acquire better technology, so that they in a more efficient way can produce their products or carry out their services. It is therefore important for companies to keep an eye on new technological innovations. The technological elements of the external environment are a part of the LoNGPEST analysis, which is being used to scan a firms environment, so it in a best possible way can adopt to it. In this essay, I will attempt to explain how technology has been a vital reason for the globalization, and how it has made the world more efficient. There is no doubt that the development of information and communications technology is one of the causes for globalization. The table below shows the reduction in communicating and travelling prices. According to the table, the price of a 3 minutes phone call have been reduced by 98,6%. The reduction in air transport costs per mile is 74%. There is no doubt that such reductions encourage companies to expand geographically. In addition, the rapid growth of the internet has made it possible to share information with the rest of the world in microseconds. Eric Samuels states that one of the industries that has grown because of information technology is the financial service industry. He states that â€Å"delivery of services has become more efficient because of the application of technology.† What he is saying, is that the financial services provided are being done more effective and at a lower cost than before. By using the internet, companies can reach out to the whole world without spending a pound. In addition to claim that the financial service industry has grown due to the information technology, Dr. Nicolas Pologeorgis also states that the industry, since 1998, has gon e through a rapid geographic expansion. This especially applies to wealthy nations, like countries from Europe and USA. He claims that â€Å"customers previously served by local financial institutions are now targeted at a global level.† Dr. Pologeorgis believes that one of the reasons for this, besides deregulation and more trading between countries, is the information technology. Services like ATMs and websites is being used to develop a more efficient relationship between long distance customers and suppliers. A study about globalization of the banking industry, states that improvements in technology â€Å"have facilitated greater geographic reach by allowing institutions to manage larger information flows from more locations and to evaluate and manage risks at lower cost without being geographically close to the customer.† One example of a company from this industry that expanded geographically in the 90’s is Goldman Sachs (GS). I 1990, Robert Ruding and Stephen Friedman decided to focus on GS global operations. During the 1990’s, GS went through a rapid geographical expansion. Today, GS has offices located in all major financial centres around the world. I have made a comparison between the growth in the financial service sector and the use of internet worldwide. It was in the 90’s that the internet use exploded. In 1995, 16 million people were connected to the internet (0,4 of world population). The table below shows that the use of internet exploded about the same time that the financial service industry, according to Pologeorgis, really started expanding geographically. However, it is not only the improvements in communication and information technology that has contributed to a more efficient and globalized world. Another industry that has been affected by improvements in technology the last century is the oil and gas industry. An article made by naturalgas states that â€Å"new innovations have reshaped the industry into a technological leader9.† A report made by the U.S. Department of energy in 1999 shows that if technology had stagnated in 1985, America would have needed twice as many wells to produce the same amount of oil and gas. However, because of technological improvement, we only need half as many wells to produce the same amount in 1999. In 1999, the America used 22 000 less wells to produce the same amount as they did in 1985. A statistical review of world energy made by BP illustrates how much oil and natural gas that has been produced in each country and worldwide from 1965-2010. Worldwide, the total development of oil has increased from 31,806,000 barrels daily in 1965, to 82,095,000 barrels daily in 2010. The development of oil has increased from 96,9 billion cubic feet daily in 1970, to 309 billion cubic feet daily in 2010. The main reason to the increase in production is off course the growing demand, but according to naturalgas it would have been impossible without the improvement in our technology. Consumption of oil increased from 30,783,000 daily in 1965 to 87,382,000 barrels daily in 2010 and consumption of gas increased from 63 billion cubic feet daily in 1965 to 306,6 billion cubic feet daily in 2010. Another industry that has developed because of new technology is the nuclear industry. Numbers taken from NEI shows that 13,5 of the world’s electricity production IN 2010, was provided by nuclear power. Furthermore, NEI states that â€Å"in total, 15 countries relied on nuclear energy to supply at least one-quarter of their total electricity†. According to NEI, one example of a country that is dependent of nuclear energy is France. 74,1 percent of the energy produced in France was generated through nuclear energy. The statistical review published by BP, shows that the consumption of electricity generated from nuclear power has increased tremendously since 1965. In 1965 the global consumption of terra hours has increased from 25,7 in 1965 to 2767,2 terra hours in 2010. FINN UT OM DAILY In this essay, I have tried to show how technology has been a vital part of both the globalization and the development of a more efficient world. I have found that information and communication technology has been important for globalization. Lower costs of travelling and communicating overseas encourage people to expand geographically. In addition, the fact that use of internet has developed into a common thing, especially in developed countries, makes customers able to do services that they could not do before. My essay also shows how the oil and gas industry have become more efficient, and how the nuclear industry has grown due to technological improvements. Reference list: Department for Business Enterprise Regulatory Reform (2008) Globalisation and the changing UK economy, page 3 [online]. Available from: http://www.bis.gov.uk/files/file44332.pdf accessed at 22.11.2011. British Petroleum (2011) Historical data [online]. Available from: http://www.bp.com/sectionbodycopy.do?categoryId=7500contentId=7068481 (then click on historical data) Accessed at 23.11.2011. U.S. Department of energy (1999) Environmental benefits of advance oil and exploration and production technology [online]. Available from: http://www.fe.doe.gov/programs/oilgas/publications/environ_benefits/4envben.pdf Accessed at 22.11.2011. U.S. Department of Energy http://www.fe.doe.gov/programs/oilgas/publications/environ_benefits/3innov.pdf Funding Universe, The Goldman Sachs group INC [online]. Available from: http://www.fundinguniverse.com/company-histories/The-Goldman-Sachs-Group-Inc-Company-History.html Accessed at 22.11.2011. Goldman Sachs (2011), who we are – at a glance [online]. Available from: http://www2.goldmansachs.com/who-we-are/at-a-glance/index.html Accessed at 22.11.2011. Internet World Stats (2008), internet growth statistics [online]. Available from: http://www.internetworldstats.com/emarketing.htm Accessed at 21.11.2011. Internet World Stats (March 31, 2011), internet usage statistics [online]. Available from: http://www.internetworldstats.com/stats.htm Accessed at 21.11.2011. Dr. Nicolas Pologeorgis (2009), The Globalization of Financial Services [online]. Available from: http://www.investopedia.com/articles/financial-theory/09/risk-free-rate-return.asp#ixzz1eNeBhksO Accessed at 20.11.2011. Eric Samuels (2011), Marketing of Financial Services [online]. Available from: http://www.measurethat.com/?cat=5 Accessed at 20.11.2011. Natural Gas (2010), Natural Gas and Technology [online]. Available from: http://www.naturalgas.org/environment/technology.asp Accessed at 22.11.2011. Nuclear Energy Institute (2011), world statistics, nuclear energy around the world [online]. Available from: http://www.nei.org/resourcesandstats/nuclear_statistics/worldstatistics/ Accessed at 22.11.2011. Wall Street Journal (2010), A Brief History of Goldman Sachs [online]. Available from: http://online.wsj.com/article/SB10001424052748704671904575193780425970078.html Accessed at 21.11.2011 Allen N. Berger, Qinglei Dai, Steven Ongen, David C. Smith (2002) To What Extent Will the Banking Industry be Globalized?A Study of Bank Nationality and Reach in 20 European Nations, page 3 [online] Available from: http://siteresources.worldbank.org/INTFR/Resources/banking_industry_globalization.pdf Accessed at 22.11.2011.

Reasons for Government Involvement in Tourism

Reasons for Government Involvement in Tourism What is tourism? Tourism can be defined in more than one way. Conceptually, it refers to the ideas and opinions people hold which shape their decisions about going on trips, about where to go, and what to do (Leiper 1995). Technically, it refers to the activities of the persons traveling to and staying in places outside their usual environment for not more than one consecutive year for leisure, health, business and other purposes (Leiper 1990, Pearce 1989). From the sociological point of view, tourism also refers to commercialized hospitality, democratized travel, a modern variety of the traditional pilgrimage, and an expression of basic cultural themes (Cohen 1984). However, the most significant form of tourism is its relations with the economic development of a country. In many countries, tourism is an economic activity which consumes a large part of its natural resources, generates revenue of billions of dollars every year and involving thousands of stakeholders and the general pu blic. As a result, it is one of the most important responsibilities of the government as to the planning, facilitating, coordinating, monitoring and protecting of tourism of that country. The World Tourism Organization (WTO) has conducted an examination on the involvement of the governments in tourism. According to Richardson Fluker (2004), a government has the responsibilities of establishing the framework of tourism, constructing infrastructure and communications; promoting the tourist destinations to the visitors, providing legislations, regulations and controls for tourism and the protecting the interests of the stakeholders. As Accrcnza (1985) pointed out there are basic stages in tourism planning which required the involvement of the government. They are analysis of previous tourism development, evaluation of the present position of tourism, formulation of tourism policy, definition of development strategy and elaboration of an action programme. All these require large amount of manpower and resources which are impossible for private sectors to manage. For instance, in establishing the framework of tourism, a functioning tourism system is identified which might affect governments decision on tourists destinations. Two major major force, demand (marketers) and supply (development), are to be considered in this respect. While the demand side is tourists who are going to the destinations, the supply side is related to the land and the geographic characteristics of the destinations (Gunn 1993). In addition, the supply side also made up of five major components, the attraction; transportation; services; info rmation and promotion. The attraction is the most powerful element as it helps to encourage and increase the desire of the tourists to visit the area. However, to develop the attraction, it may also involve the considerations of such external factors as the natural resources and the cultural resources. The natural resources provide the foundation for tourism development which preferred by tourist, such as water life; vegetative cover; wildlife; topography, soils, geology and climate, atmosphere. The Cultural resources are important for attractions developed as historic; ethnic national, craft and related economic development. As a result it may require the government to win the support of the community regarding the development of these destinations as the community is a significant factor on the functioning of tourism. Its acceptance and willingness to support tourism industries would greatly facilitate the long term development of the sites. Transportation which enable tourist to travel to different destinations; and services and information such as shops, banks and restaurants; are two other components of the supply side. As Gunn (1993) pointed out it was the responsibility of the government to create new and improve physical plants such as superstructure and infrastructure. Superstructure refers to the land and building developments used directly by travelers, such as hotels, restaurants and attractions. Infrastructure is applied to necessary support developments, such as roads, airports, water supply and fire protection. These public facilities have to be provided by the government so as to promote the growth of tourism. In addition, (Gee, Dexrer James 1984) physical development of tourism is made up a great diversity for business, government, and nonprofit components, such as direct provider, include transportation, travel agencies and restaurant; the support services which are tour organizers, management; and the developmental organization include planners and government agencies. Regarding the promotion of the tourist industry which helps to increase tourist to visit that area, government agencies have an important role as the promoter in this industry. Advertising in mass media and internet is very important in promoting and stimulating travel to the destinations. Familiarization tours and travel shows can provide information and attract tourist to visit that destination. Moreover, if they are organized by the government, they will become promises with reputation and will increase the numbers of the visitors significantly. Tourism product such as parks, historic sites or convention centers can increase the tourist flow, but these attraction products must be aware as a promised. If tourists discover the destination is not the same as its promise, it may affect the number of tourist who willing to come to the destination (Gunn 1993, Pearce 1989). Therefore, management of these tourism products has to be carefully supervised and monitored. Management is to make institutions capable of producing result (Drucker 1998). According to Richardson Fluker (2004), the management method which adopted by the WTO are the consulting with various stakeholders through meetings with industry, advisory groups and the general public The aim of these meetings is to discuss specific subjects such as strategic planning; issues on tourism industry such as promotions and regulations; setting up and funding of specialized tourism agencies which help to develop the tourism industry, and providing facilities and operations in ord er to provide money or facilities for national or communities interest. Moreover the meetings are also about creating the appropriate fiscal, regulatory and social environment within private business; and adjudicating between conflicting interest on society which are concerned with the environment, recreation and wildlife recourses. As we can see, management is also related to the protection on destinations interest. It is very important in tourism industry. Decision-making through managerial perceptions and the objective dimensions of tourism industry structure influence the natural environmental resources. The tourism resorts and its environment are necessary for survival and growth. The strategic managers have the most responsibility to that environment. As a result, the planning process in tactical (short-term) and strategic (long term) has to be carefully considered and monitored as it can improve the tourism industry performance and allow an effective response to a continually changing environment (Knowles, Dianmantis El-Mourhabi 2004). All in all, the development of tourism of one country is closely related to the policies of that government. According to Richardson Fluker (2004), policy is a course of action adopted and pursued by a government, expressed as a broad statement of general intent which guides the actions of public servants and others in carrying out the governments wishes. In other words, only the government has the power to command (including by legislature) such an enormous resources to develop strategy which matches the demand and supply factors such as an evaluation of tourist and market resources; the visitor satisfaction; environmental protection and adequate rewards for developers and investors. In conclusion, the government plays an important role in the tourism industry. Government agencies have to provide legislations, regulations and controls for tourism in order to ensure its tourism industry can sustain its level and compete with other tourist destination. They have to establish a framework which ensures its tactical and strategic in the attractiveness within the tourist destination in order to increase the amount of tourist visitation. The protection of interest is another important factor of the destination area needed to be considered too. If the attractiveness of its destination decline, it will affect the numbers of tourists who are willing to visit. As a result, to maintain a stable attractive in tourist destination, a new destination may have to be explored or artificial attract resources may be needed. Government agencies have to be concerned with the acceptance in the society whether they welcome or in conflict with the tourist interests. Government agencies need to make a good balance between two important factors. The promotion on tourist destinations helps its national economic and attraction competitiveness around the world. Tourists who are attracted by the promotion will come to visit instead of other places, so that the national economics will increase. However, if the destination doesnt match with what the government agencies promote, the national reputation may decline. As a matter of fact, government agencies play an important role in the tourism industry as the planner, coordinator, legislator and regulator, stimulator, promoter and protector of the destination interest.

Role of Elevated Levels of Homocysteine in Blood Plasma

Role of Elevated Levels of Homocysteine in Blood Plasma Homocysteine, a non-protein amino acid, is an intermediate in the metabolism of methionine and biosynthesis of cysteine. It has gained prominence in the past half-century because its accumulation in the body has been linked to increased risk and occurrence of atherosclerosis and cardiovascular disease. Homocystinuria is due to errors in metabolism and homocysteinemia is attributed to polymorphisms in the genes involved in methionine metabolism and cysteine biosynthesis, and deficiencies in the nutrients folic acid, and vitamins B12 and B6. Proof of these can be found in the results of genetic and dietary studies. The clinical quantification of homocysteine has evolved through the years and routine tests are currently available. Baseline homocysteine concentration for normal adults has been identified to be between 12-15 Â µmol/L. Elevated levels of homocysteine can be reduced with folate and B vitamins supplementation, but this intervention does not work in patients who have suffere d stroke or heart attacks. More research studies have supported the hypothesis that elevated homocysteine causes CVD and is not just a marker for the disease. However, despite the amount homocysteine research performed, many issues remain to be resolved, among which are the elucidation of the molecular mechanism of the direct action of homocysteine and the standardization of techniques for the quantification of homocysteine levels. Brief introduction. 200 w The role of elevated levels of homocysteine in blood plasma has been the subject of intense study and literature reviews for more than 50 years since an association between defects in homocysteine metabolism and thromboembolism was observed. This role was further verified in patients with homocystinuria and abnormalities in vitamin B12 metabolism with general vascular damage and widespread thrombosis. Subsequently, the homocysteine theory of arteriosclerosis was formulated by McCully and Wilson in 1975. Since then, the positive association between the risk of cardiovascular disease and homocysteine levels in the general population was validated in many epidemiological studies (Boushey, et al., 1995; Verhoef, et al., 1996; Eikelboom et al., 1999; Humphrey et al., 2008). Significantly, these studies found that small increases of homocysteine levels in blood increase the risk of coronary heart disease. Another important finding was the role of diet, vitamins and folic acid in lowering h omocysteine levels. Some authors have questioned the direct homocysteine-cardiovascular disease link, basing their conclusions on a review of longitudinal, prospective studies (Kaul, Zadeh and Shah, 2006). Accordingly, the evidence is not strong enough to warrant a causal effect, the mechanisms for how homocysteine causes cardiovascular disease has not been elucidated, and that there has been no proof showing that interventions of decreasing homocysteine levels have modified the risk for atherosclerosis (Kaul, Zadeh and Shah, 2006). Moreover, homocysteine has been proposed to be a marker, and not a cause of CVD (Wierzbicki, 2007). Recent studies have proposed a mechanism on increased risk of cardiovascular disease (CVD) due to elevated homocysteine. Inhibition of the growth of endothelial cells and promotion of vascular smooth muscle cell proliferation results in damage of vascular cells (Zou, 2007). The abnormality in the production of endothelial cells was caused by homocysteines inhibition of DNA methylation in the promoter region in the gene of an activator of the cell cycle (Jamaluddin, et al., 2007). Amino acid and focus on cysteine. 300 w All living cells contain the biomolecules proteins, carbohydrates, nucleic acids and lipids (Mathews and Van Holde, 1996; McKee and McKee, 2004). These are made up of the repeating sub-units of amino acids, sugars, nucleotides and fatty acids respectively. Cells also contain small organic molecules that are involved in complex biosynthetic and regulatory pathways that are tightly controlled at the molecular level. Amino acids are naturally occurring compounds containing an amino group, a carboxyl group, and a unique side chain or R group (Figure 1). The chemical and functional properties of an amino acid are largely determined by its R group (McKee and McKee, 2004; Mathews and Van Holde, 1996). Although there are hundreds of amino acids, only twenty have been identified to be building blocks of proteins. Among these is cysteine, which contains a sulfhydryl, or thiol (SH) in its R group (-CH2-SH) (Berg, Tymoczko and Stryer, 2002). The sulfhydryl group is very reactive and could react with another thiol group, forming disulfide bonds or sulfide bridges. These bridges are important in increasing the stability of some proteins. Two sulfhydryl moieties of two cysteine bonds can oxidize forming cystine. This commonly occurs in extracellular fluid like blood and urine. However, the solubility of cystine is very low, and in large amounts, cystine can cause kidney stones (McKee and McKee, 2004).The sulfhydryl moiety also forms weak bonds with nitrogen and oxygen. Another amino acid which contains a sulfhydryl group is methionine. Its side chain is -CH2CH2SCH3. The sulfur in methionine can form bonds with electrophiles (Berg, Tymoczko and Stryer, 2002). The methyl group (-CH3) can be activated and is involved in many reactions where just one carbon atom is being added to another compound. Cysteine is considered a non-essential amino acid because it can be synthesized de novo in both plants and animals (Figure 2). In animals, cysteine is formed from the cleavage of cystathionine through the action of ÃŽ ³-cystathionase. Cystathionine is produced when serine condenses with homocysteine that is derived from methionine (McKee and McKee, 2004). Methionine, an essential amino acid, cannot be synthesized in humans, and thus, have to be provided in the diet. Its metabolism involves homocysteine. Homocysteine structure and metabolism. 200 w Homocysteine is a non-protein amino acid and thus, it is not utilized in protein synthesis. Its structure is similar to cysteine but it has an additional carbon before the sulfhydryl moiety (R group: -CH2 -CH2 -SH). The tendency of homocysteine to form cyclic compounds may have limited its potential as a protein building block. Homocysteine is not provided in the diet, but is biosynthesized from methionine in a process involving several steps. Methionine is first adenosylated to form S-adenosyl methionine (SAM), and then the methyl group is transferred to an acceptor molecule to form S-adenosyl homocysteine (SAH) in a process called transmethylation. Adenosine is then removed thereby forming homocysteine (Figure 3) (Durand et al., 2001; Selhub J. , 1999; Marinou, et al., 2005). Transmethylation is the only pathway for producing homocysteine in the body. Homocysteine then undergoes two pathways: remethylation to methionine, and transsulfuration to produce cystathionine, cysteine, pyruvate and taurine. Remethylation requires folate and cobalamin (vitamin B12), and transsulfuration requires pyridoxine (vitamin B6). Homocystinuria results from genetic errors in the metabolic pathways (Kluijtmans, et al., 2003; Klerk, et al., 2002), but the current focus is on the more common hyperhomocysteinemia that results fr om genetic variations or nutritional inadequacy (Finkelstein and Martin, 2000). Polymorphisms in the gene methylenetetrahydrofolate reductase (MTHFR) which catalyzes the transfer of a methyl group to homocysteine to re-form methionine have been found in many studies to increase homocysteine levels (Klerk, et al., 2002). Among the proposed effects of increased homocysteine levels resulting in CVD are oxidation of low density lipoprotein, decrease in the thrombomodulin expression inhibiting the anticoagulant pathway leading to thrombosis, platelet activation and aggregation, and smooth muscle cell proliferation (Eikelboom, et al., 1999; Jamaluddin, et al., 2007). Normal Homocysteine levels. 200 w The concentration of circulating total homocysteine (tHcy) is an accurate marker of low folate and vitamin B12. In 1999, 3563 male participants and 4523 female participants were surveyed to determine the normal blood homocysteine levels (Selhub, et al., 1999; Selhub, 1999). The survey found that homocysteine increased with age and was higher in adult males than and females. Screening of babies confirmed that homocysteine levels are higher in baby boys (Refsum, et al., 2004). It was recommended that blood homocysteine levels equal to or greater than 11.4 Â µmol/L (males) and 10.4 Â µmol/L (females) are to be considered above normal and was associated with low vitamin concentrations in two-thirds of the subjects surveyed (Selhub, et al., 1999). This verified that an assessment of homocysteinemia requires the knowledge of the health status of the patient. The degree of homocysteinemia is classified based on determination fasting levels of serum homocysteine. Currently, a plasma tHcy level of 15 Â µmol/L is considered as normal (Refsum, et al., 2004). In healthy adults with good folate and B vitamins status, the upper reference limit is 12 Â µmol/L. Accordingly, homocysteinemia is classified as moderate (15-30 Â µmol/L), intermediate (30-100 Â µmol/L) and severe (> 100 Â µmol/L). This categorization is necessary to come up with a decision regarding the treatment of the condition. Homocysteine and nutrition. 300 w Studies have shown that the enzymes regulating the metabolism of homocysteine are activated by the B vitamins and folate which can be sourced from the diet. The balance of the remethylation and transmethylation pathways are coordinated by nutritional inputs, specifically methionine (Selhub, 1999). Decreasing dietary methionine increased homocysteine remethylation, while increased methionine increased the production of cystathionine through upregulation of the transsulfuration pathway. The molecular mechanisms for the effects of dietary methionine are based on the capacity of S-adenosyl methionine SAM to inhibit methylenetetrahydrofolate reductase (MTFHR) and activate cystathionine synthase (Selhub and Miller, 1992). Thus, when dietary methionine is high, there is a rapid conversion of methionine to SAM. The high levels of SAM promote the inhibition of MTHFR and subsequently, remethylation of homocysteine is also depressed. The pathway is channeled to the transsulfuration pathway, to produce cysteine. In opposite conditions of low dietary methionine, low SAM levels are not enough to inhibit MTFHR activity, resulting in the remethylation of homocysteine. The role of folate in homocysteine metabolism has been the subject of many research studies (Antoniades, et al., 2009; Boushey, et al., 1995; Lonn, et al., 2006). A diet that is poor in folate was also found to impair remethylation and synthesis of SAM (Miller, et al., 1994). Folate deficiency increased the concentration of tHcy and hepatic SAM concentration. Introducing dietary folate and methionine also decreased tHcy. Since vitamins and trace minerals are lost due to food processing, fortification is proposed in order to replenish the lost nutrients. Folic fortification of breakfast cereals when consumed daily was recommended to be the most effective means of decreasing serum folate (Riddell, et al., 2000). High-dose folic acid supplementation also decreased tHcy levels in cases where there is vitamin B12 deficiency (Min, 2009). However, a high methionine diet is not recommended since it can decrease body weights and HDL-cholesterol production in mice (Velez-Carrasco, Merkel and S mith, 2008). Aside from the observed effects of folic acid on reducing tHcy concentrations, folic acid is also thought to be involved in ameliorating endothelial dysfunction through its action of maintaining endothelial nitric oxide synthase in its coupled state which favors the formation of nitric oxide, and not oxygen radicals (Moens, et al., 2008). Role of Homocysteine and vitamins. 200 w The enzymes involved in metabolism of homocysteine require the participation of vitamin cofactors B6 and B12, and folic acid. The direct association between low folate, vitamin B6 and B12 status and hyperhomocysteinemia have been validated many times Hao, et al; 2007; (Hao, et al., 2007; Boushey, et al., 1995; Kluijtmans, et al., 2003; Koehler, et al., 2001). Investigations have geared towards the probability of using these vitamins in the prevention and treatment of homocysteinemia in the general population, and in those who already have cardiovascular disease. The use of multivitamins supplementation for several months improved the concentration of the B vitamins and folate acid in plasma, and decreased homocysteine and LDL cholesterol levels in 182 study participants (Earnest, Wood and Church, 2003). Similar homocysteine-lowering effects were also observed in patients with celiac disease, who have malabsorption problems, who take vitamin supplements (Hadithi, et al., 2009). However, the case for the vitamins is different in patients who already have CVD. Although the treatment of different combinations of vitamins B6, B12 and folic acid of patients with coronary artery disease resulted in a 30% reduction of tHcy one year after receiving vitamin B12 and folic acid, follow-up measurements of homocysteine levels showed that the vitamin supplementation did not significantly affect the total cardiovascular events (Ebbing, et al., 2008). Thus, the findings do not support the use these vitamins for secondary prevention of coronary artery disease. The Norwegian Vitamin Trial (NORVIT) also found that vitamin supplementation did not reduce the risk of cardiovascular disease recurrence after a heart attack (BÃ ¸naa, et al., 2006). Moreover, preliminary results of the NORVIT suggested that the B vitamin treatment increased risk of cancer, and that such treatment should not be given. Role of Homocysteine in atherosclerosis. 500w Many studies since the 1960s have validated the relationship between hyperhomocysteinemia and risk of atherosclerosis (Eikelboom, et al., 1999). Severe hyperhomocysteinemia (homocysteine levels greater than 100 Â µmol/L) can be caused by several inherited genetic disorders. Foremost among the disorders are mutations in the gene encoding for the major transsulfuration enzyme cystathionine ÃŽ ²-synthase, remethylation enzymes methylenetetrahydrofolate reductase, methionine synthase in the methionine cycle, or defects in vitamin B12 metabolism (Finkelstein, 1998; Kraus, 1998). These genetic conditions lead to extreme elevations of plasma homocysteine and early atherothrombotic disease, where the typical pathologic features of endothelial injury, vascular smooth muscle cell proliferation, and progressive arterial stenosis are observed. While these genetic errors in metabolism are rare, they gave researchers a model for studying cardiovascular injury that was associated with high homocys teine levels. The clinical and pathologic features observed under the conditions of homocystinuria resulted in the homocysteine theory of atherosclerosis that declared high plasma homocysteine concentrations to be responsible for vascular damage. Results of individual studies and meta-analysis have come up with support for the homocysteine theory of atherosclerosis. However, there are still controversy over whether homocysteinemia causes CVD or vice versa, or is it that homocysteinemia is just a marker or indicator of CVD. Mounting evidence for the causal effect of homocysteine was backed up by basic and cellular studies which utilized advanced molecular and genetic techniques. Early studies infer that homocysteine damages the endothelial cells lining the blood vessels and increase the production of vascular smooth muscle (Berg, Tymoczko and Stryer, 2002). From a pathophysiologic point of view, homocysteinemia is associated with increased oxidative stress in the cells, and development of thrombosis (Tyagi, et al., 2005), impaired endothelial function (Stuhlinger, et al., 2001) and the activation of inflammatory pathways that are sensitive to changes in cellular redox states (Weiss, et al., 2003). It was originally proposed that homocysteinemia promotes the accumulation of S-adenosyl homocysteine, a potent inhibitor of cellular DNA methylation (Zou, 2007). The hypomethylation of DNA could affect promoter activity resulting in remodelling of chromatin and changes in the transcription of certain genes. However, it was found that In the case of homocysteinemia, increased tHcy levels cause the hypomethylation of DNA promoter region of the cyclin A gene (Jamaluddin, et al., 2007; Zou, 2007). Cyclin A protein is a sub-unit of cyclin-dependent kinases (CDK) that activates CDK during the cell cycle. In the presence of high tHcy concentration, methylation was inhibited at two CpG sites in the cyclin A promoter, resulting in a 6-fold increase in promoter activity, and therefore increased proliferation of endothelial cells. Homocysteine directly inhibited the activity of DNA methyltransferase I (DNMT1) by 30% (Jamaluddin, et al., 2007). Furthermore, homocysteine reduced binding of methyl CpG binding protein 2, while it also increased the binding of cyclin A promoter to histones H3 and H4 which led to chromatin remodelling. Review methods of analysing homocysteine and differences between measurements. 2800 w Total homocysteine, tHcy, is the sum of all chemical species of homocysteine that is present in the blood (Figure 4) (Ueland, et al., 1993; Refsum, et al., 2004). As mentioned earlier, only a small portion of the tHcy comes from free homocysteine, a large portion comes from mixed disulfides and protein-bound homocysteine. All the assays used to quantify tHcy cleave and convert all these forms into a single species by adding a reducing agent, therefore what is measured is the reduced form. Blood for homocysteine determination is collected in tubes. If plasma homocysteine is to be measured, the tube contains anticoagulants like EDTA, heparin or sodium citrate. If serum homocysteine levels are to be determined, then the samples are allowed to coagulate, which takes thirty minutes and not more, to reduce the risk of increased homocysteine levels coming from the cellular fraction of blood (comprised of red blood cells and platelets). Increased release of tHcy from cells also makes it necessary to centrifuge and separate blood components within 30 minutes of collection. Centrifugation time and speed are relatively low (5 min and 300g respectively, for example) and is performed at 4Â ° C. The supernatant (serum/ plasma fraction) that is collected may be analyzed immediately, or stored for 2-3 weeks in the refrigerator or frozen for 2-3 months (Homocysteine, 2008). It is possible to store the samples once processed because the homocysteine levels have been shown to be stable after separation of plasma/serum from the cellular fraction (Refsum, et al., 2004). There are reports showing that tHcy in serum can remain stable for several years if stored at -70 Â °C. Scheme for the steps followed in the determination of total homocysteine from blood serum or plasma. (Hcy-SR, Hcy-mixed disulfide; EC, electrochemical detection; LC, liquid chromatography; CE-LIF, capillary electrophoresis with laser-induced fluorescence detection; Ab, antibody; EIA, enzyme immunoassay; FPIA, fluorescence polarization immunoassay; MS-MS, tandem mass spectrometry). Figure from Refsum et al., 2004. The first step is to generate free homocysteine by chemical reduction of disulfide bonds using dithioethritol, sodium borohydride, n-tributyl phosphine, 2-mercaptoethanol and phosphine tris (2-carboxyethyl) phosphine, which is water-soluble (Jacobsen, 1998). After the reduction, homocysteine is separated from the cysteine, glutathione and cysteinylglycine, low molecular weight thiols, by HPLC. Homocysteine can also be derivatized for gas chromatography determination/ HPLC or subjected to immunoassay (Jacobsen, 1998; Refsum, et al., 2004). The development of the methodologies for determining total homocysteine started in the 1980s. There are many variants of the methodologies, but they can be categorized into three general methodologies: (1) chromatographic, (2) enzyme immunoassays and, (3) enzyme cycling. Chromatographic methods The earliest methods used were the chromatographic methods, of which high pressure liquid chromatography (HPLC) is more used compared to gas chromatography (GC). Variations in the methodology include the combination of liquid chromatography (LC) with mass spectrometry (MS) and GC with MS. The equipment and the skill requirements for these techniques have limited their use mainly in research laboratories. In the HPLC method, the reduced samples are deproteinated and then directly separated on an ion exchange column. The separated homocysteine molecules undergo derivatization with ninhydrin (a chromophore for colorimetric quantification), or fluorescent compounds (e.g. fluorescamine, SBD-F). Alternatively, the reduced homocysteine molecules are first derivatized before separation through reversed-phase HPLC. 6-Aminoquinolyl-n-hydroxylsuccinimidyl carbamate (AQC) is a fluorophore that gives higher sensitivity when used to derivatize cystathionine and homocysteine (Seo, 2005).The derivatized samples are then passed through an ion exchange column. The length of time that it takes for the sample to go through the column is called the retention time. The samples leaving the column pass through a detector, which could be a UV absorbance detector (spectrophotometer), a fluorescence detector or a mass spectrometer which is a powerful detector. The detectors quantify the samples, and send signa ls to a recorder which produces the chromatogram, where quantities can be seen as peaks. An internal standard of known quantity is used in the determination of the actual quantity of the sample. Normally, the internal standard should have a retention time that is the same as homocysteine. Internal standards are added to the sample tubes and derivatized together with the reduced homocysteine. To quantify homocysteine, the tHcy peak area is compared with the peak area of the standard eluted at the same retention time. Although there are many advancements in the HPLC technology, tHcy quantification using this method is relatively slow, time-consuming, requires very careful sample preparation and handling, and skilled technicians. Sample pre-treatment requires 30-60 minutes, while an HPLC run takes 10-30 minutes. Samples are run sequentially, which makes HPLC not suitable for high throughput determinations. Some laboratories use gas chromatography or gas chromatography with mass spectrometry to get more sensitive results. The principles behind GC and HPLC determination are similar, except that in GC, the samples are vaporized and passed through a gas before elution from a column. Similar to HPLC, GC is also more time-consuming compared to other methods. Homocysteine of dried blood samples have been determined using reversed phase HPLC (Bowron, et al., 2005). In this technique, blood samples were collected in tubes that contain potassium EDTA. After mixing gently, the blood was spotted on filter paper cards that are used in standard neonatal screening. The cards were air-dried and stored at room temperature until analysis. Six-mm discs of the cards were punched out, and incubated in tubes with an internal standard and a deproteinization agent. The produced thiols were then derivatized for fluorescent detection, and separated by reversed phase HPLC. The resulting measurements showed that the method was linear up to a homocysteine concentration of 140 Â µmol/L, in comparison with HPLC of plasma homocysteine which showed a linearity up to 200 Â µmol/L. The coefficient of variation within a batch of measurement was 5% and within batches was 8%. HPLC of plasma samples was only 1% within batch and 4% between batches. Storage of the dried blood sample spots for more than 24 hours in room temperature decreased the homocysteine concentration, probably due to the hemolysis of whole blood during drying. The hemolysis could have been accompanied by the denaturation of enzymes that release homocysteine from blood cells. Overall, although the use of dried blood samples offers flexibility in sampling and storage, the assay had lower precision compared to HPLC. It may not be able to quantify dried blood homocysteine in newborns because its lower limit of detection is above the levels that some healthy neonates have. The method can be used in the detection of homocysteinuria. A more rapid protocol for tHcy measurement is liquid chromatography electrospray ionization-tandem mass spectrometry (LC/MS/MS) (Gempel, et al., 2000). It removes the derivatization step and can perform 400 analyses daily. Blood samples are applied to filter cards that are used for neonatal homocysteine screening. The cards are incubated in the presence of a reducing agent and a deuterated internal standard. The tHcy is then extracted in an acetonitrile solution and loaded on a cyano column. Eluates are ionized with an ion spray device before passing through a mass spectrometer. The procedure makes use of an autosampler, which hastens the analysis. The advantage of the method is the removal of the derivatization step, because the mass spectrum is determined directly. Impurities of the samples are removed by passing them through the cyano column. Since the procedure takes only 3 minutes to finish 1 sample, more samples can be processed in less time compared to HPLC (Gempel, et al., 20 00). An innovation in a high throughput liquid chromatography-tandem mass spectrometry method is the use of a 96-well plate format that skips precipitation, centrifugation and derivatization. Plasma samples, calibrator and control were mixed well in 96-well plates. From each well, an aliquot was transferred to another plate with sodium hydroxide solution (diluent) and dithioethritol (or another suitable reductant). After thorough shaking and a short incubation period, the plate was placed in the autosampler for LC-MS/MS analysis. Comparison of the results with those obtained by HPLC showed that the LC-MS/MS method gave consistently higher homocysteine readings, which was attributed to the use of different standards by the two methods. The biggest advantage of the new method over HPLC was the expense, because LC-MS/MS reduced the costs for material and manpower by more than 50% (Arndt, et al., 2004). Enzyme Immunoassay The immunoassay-based tests for homocysteine were developed in the 1990s. Most assays employ a mouse monoclonal antibody against S-adenosylhomocysteine (SAH), which is formed when adenosine and S-adenosylhomocysteine hydrolase are added to reduced total homocysteine of the plasma/serum sample. The earliest report on the procedure involved the use of flouresceinated SAH as tracer. This method is fluorescent polarization immunoassay or FPIA. After SAH is generated, it is mixed with a fluorescein-tagged SAH. The mixture is coated onto a microtiter plate, then made to bind with mouse anti-SAH (Figure 6). When antibody-SAH bind, the changes in fluorescent polarization of the SAH-tracer are detected and serve as basis for the quantification after a standard curve of known homocysteine calibrators is constructed (Shipchandler and Moore, 1995). The newer versions of enzyme immunoassays were performed on microtiter plates and used the formation of colored complexes to measure absorbance which was used as the basis of the quantification (Figure 7) (Frantzen, et al., 1998). After the conversion of reduced tHcy to SAH, another enzyme is added to the mixture to hydrolyze the excess adenosine. After reaction with a monoclonal anti-SAH antibody, a specific anti-mouse-antibody is added, which forms a colored product that can be detected spectrophotometrically for quantification. The immunoassays are simpler to perform compared to chromatography because they do away with the derivatization step and the expensive equipment needed in chromatographic separation. The method has been proven to be rapid and precise. The coefficient of variation between and within assay is 8% and less than 6% respectively, and the results also correlated well with those obtained using HPLC. The advantage of the immunoassay over HPLC is speed and ease of determination (Frantzen, et al., 1998). Many samples can be run simultaneously on several microtiter plates, which make the immunoassay ideal for routine screening. Furthermore, the technique can be and has been fully automated using robotics technology, and is highly flexible and can be used on different analytical platforms. Other similar imunoassays have been developed. In a variant of the assay, recombinant homocysteine lyase, converts homocysteine into ÃŽ ±-ketoglutarate, with the evolution of ammonia and H2S (Tan, et al., 2000). A second reaction reacts H2S with N,N-Dibutylphenylene diamine (DBPDA) resulting in the formation of form 3,7-bis(dibutyl amino)phenothiazine-5-ium chloride, a highly fluorescent compound. Both highly purified recombinant homocysteine lyase and DBPDA were synthesized in the authors laboratory, which somehow limits the utility of the procedure because their production requires skill and specialized equipment. Another assay utilizes the recombinant enzyme, methionine ÃŽ ³-lyase, and the commercially available fluorophore N,N-diethyl-p-phenylenediamine (Chan, et al., 2005). The rest of the procedure is similar to that used in typical immunoassays. The method can be used in an automated analyzer or manually, which makes it ideal for laboratories that cannot afford the expensive automated analyzers. Although the recombinant enzyme was purified only from a crude extract of E. coli, the results compared well with those that were obtained using HPLC. These results showed that the need for tedious and careful purification procedures can be removed. However, a main drawback was the non-specificity of the enzyme for homocysteine; it also reacts, although with less specificity, with cysteine which is present in larger amounts than homocysteine in the plasma. Thus, the use of pure homocysteine as calibrator is not suitable for the assay. Instead, the recommendation is to use pooled plasma as the cali brator, but this requires the determination of the tHcy by HPLC, which complicates the procedures for small laboratories who do not have access to HPLC equipment. Moreover, the precision of the procedure is also lowered if samples are diluted, because this also reduces the amount of cysteine in the sample relative to its concentration the calibrators. The presence of cysteine is also the cause for the lower sensitivity of this assay, because at lower tHcy levels the background fluorescence due to cysteine is amplified (Chan, et al., 2005). Enzyme cycling The most recent method for tHcy determination is the enzyme cycling method which amplifies the detection signal to improve the sensitivity of the detection (Figure 8) (Dou, et al., 2005). Similar to the previous methodologies, all protein-bound and oxidized forms of homocysteine are first reduced to free homocysteine. Next, methionine and S-adenosyl homocysteine (SAH) are formed from the methylation of free tHcy by the action of the enzyme homocysteine methyltransferase with S-adenosyl methionine as methyl donor or co-substrate. The SAH so formed comes from SAM and the transmethylation reaction, and not from the free homocysteine molecules. Upon formation, SAH is hydrolyzed immediately to homocysteine and adenosine by SAH hydrolase. The homocysteine originating from SAM enters the homocysteine transmethylation reaction catalyzed by homocysteine methyltransferase to form homocysteine and methionine again, resulting in an enzymatic cycle and an increase in detection signals. The reacti ons caus