dr. baharuddin badi

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  • Summary Report of Activity Student Exchange Program (MEXT PROJECT) Collaboration Airlangga University (Indonesia) and Kobe University (Japan)

    & Exchange Program Faculty of Health Science, Kobe University Myodani Campus, Kobe, Hyogo Prefecture, Japan 2013

  • ReportofActivitySupervisor :Prof.HakHottaSensei :MasanoriKameoka,PhDNameofStudent :BaharuddinBadi,S.Si(equivalentB.Sc)Introduced of Instruments andMediumwhich used for Cell Culture andDengue VirusExperiment:Purpose : For knew all instruments and understand how to operations thatinstrumentsbysafely,duringexperimentandalsoknewthekindofmedium incellcultureandDengueVirus(DEN)experiment.Results :

    1. RPMI1640MediumfromSigmaAldrich:Descriptionscompositionsare:RPMI1640Medium(500ml)+FBS10%+1%PenicillinStreptomycin.ThisisthemediacompositionforK562cellsculture.

    2. Minimum Essential Media (MEM), the media powders is made by Nissui

    Pharmaceutical Co., Ltd.:

    Descriptionscompositionsare:MediumMEM500ml+FBS10%+1%LGlutamine+AdjustpH

  • Conclusions :1. RPMI1640MediumfromSigmaAldrich:isverygoodtoappliedforcellculture,in

    thiscaseisK562cell(semiadherenttype),becausethismediahascompoundmacroandmicroelementsuchas:Inorganicsalts,Aminoacids,Vitamins,andCarbohydrate(Dglucose).

    2. The MEM (Minimum Essential Media) Media from Nissui Pharmaceutical Co., Ltd.:isverygoodtoappliedforcellcultureinthiscaseisVerocell(adherenttype),because this media has compound macro and micro element such as: Sodiumchloride,Aminoacids,Vitamins,Carbohydrate(glucose)andKanamycin.

    3. AllInstrumentsandMaterialsuchasmediumbeforeandduringwasworkingoncellculture must be sterile and condition must aseptic for prevent there arecontaminations.

    CellCultureExperiment:Purpose :

    1. ToknowhowtomakingmediaforK562cellandverocell.2. Thepurposeforthissessionistorejuvenate,growthupandincreasedthenumbers

    ofcells.Results :

    1. PicturesofVerocells.

    Descriptions:

    A. Verocelltheageof3dayswithconfluent50%.B. Verocelltheageof5dayswithconfluent70%.C. Verocelltheageof6dayswithconfluent100%.

    2. PicturesofK562Cells.

  • Descriptions:A. Verocelltheageof2dayswithconfluent40%.B. Verocelltheageof3dayswithconfluent90%.C. Verocelltheageof5dayswithconfluent100%.

    Conclusions :

    1. The Vero cell hadwe used is a cell linewhich isolated from kidney epithelial anAfricangreenmonkeysbyscientists.Thiscellistypeadherentiscausedverystrongtoattachedonpolystyrene TissueCulture Flask Falcon. Sobecauseof thatweusedTrypsinEDTAhassterilizedwithfilter,fordetachedit.ForcultureofVerocellweusedMEMmediawithcompoundofmaincompositions is :MEM500ml+FBS10%+LGlutamine1%+AdjustpHwithNaHCO3(46ml)andtheresultisgood.

    2. The K562 cell hadwe used is a cell line from erythroleukimia and this is a semiadherent type itcausedcouldnotstrong toattachedonpolystyreneTissueCultureFlaskFalcon such as T75 or T25.

    3. Thegrowing rateofcell iscausedofmany factors suchasconditionofmedia.Thecompositionseveryelementsinmediamustappropriatenecessaryofcell.Afterthatthesecondimportantisconditionsduringincubationsperiod,inthiscasepointingatCO2 concentrationsand the setof temperatures, if there is imbalance thegrowingrateofcellwouldbecometolowerandcontinuetodie.

    DengueVirusInfectionsExperiment:Purpose :

    1. ForunderstandaboutCPE(CytophaticEffect)whileDengueVirusisinfectedtoVerocell.

    2. ForgettingmuchmoreofDengueVirusquantity.3. Thefirstactivityforcountthetiterofvirusobtainedlasttimeharvestedprocess.

    Result :

    1. The Vero cell which experienced of CPE (Cytophatic Effect) because infected ofDengueVirus(DEN).

  • 2. The solution which containing Dengue Virus from Harvesting process with anadditionalNaHCO37,5%foradjustpH.

    Conclusions :

    1. TheCPECytophaticEffectwecansee itonsixthormoredays.Thewrinklingthatwas IndicationofCPEonthecellsthis iscausedofmembranewasbeingdamagedandifthisinjuryhasseveredcanmakeacelltodie.

    2. Harvestingprocesswedoneon fourth,sixthandeighthday.Allsolution fromT75which compound of Dengue Virus taken and put into 50 ml Falcon tube andcentrifugedwithused3.000rpmwithin5min.Afterthat,addedNaHCO3foradjustpHandsaveinfreezer800C.

    Immunostaining:Purpose :Forknewatiterratelevelinoursamplesothatwecanmakeappropriatedilutions.Result :OneofresultstainingprocessforDengueVirusformakingatiter.

  • Descriptions:TheresultofstainingprocessonM24platesby lasttimewhich incubationofVerocelland infectedwithDengueVirus(onpictures isshowedDEN1day4,6andDEN4day6and8).Conclusions :Countingofvirustiter.CalculationsperformedonthevirustiterplateM24(Falcon)withavolumeof1ml@wellis,whopreviouslyperformedmultileveldilutionordilutionbyusingplateM96 (Falcon ). This procedure is performed to determine the titer of virus stockowned.OfthisprocesscanalsonotethatthesizeoftheplaquethatformsonanytypeofviruscanbedifferentwheretheimagesizeissmallerplaqueDEN2comparedwithDEN1.ElISAforDengueVirusExperiment:Purpose :

    1. ForchecksthereareornoDengueVirusonsamples.2. Forcorrectionwhetheranantigenhasappropriatetoantibody.3. Tocompareswithdenguevirusdilutions.

    Result :

    1. ResultfromELISAprocesswhicheventreactionsantigensandantibodies.Conclusions :FortheELISAtestweusedMaxiSORPplate96NUNCTMbecausethebottomof the layer (stationary phase) can strongly capture antibody. For antibody (capture)weusedantiNGCdengue(DEN2)Thisprocess isknownas"coatingantibody"afterthatweadded antigen in this case is dengue virus DEN 2 NGC strain (New Guinea Strain) andprimary antibody (primary detector)we added this is an universal antibody for dengue(4G2),(IgG)whichisan"ascitesfluid"orMAbfrommice.Thenextstage,addedasecondaryantibody(antimouseIgG)alkalinephosphataselabeledsubstratePNPPthenandincubated,andreadonamachinereaderinexperimentedweusedinstrumentfromBioRad.

  • 2. ResultfromreaderELISAprocess.

    Conclusions :WithusingReaderInstrumentfromBioRadwecanseetheresdecreasedofquantityof DengueVirus (DEN)which indicated lowofOD (opticaldensity)valueneartozero"0"pointwhendilutionupper109specialforDEN1day8andDEN2day8.

    WesternBlotExperiment :Purpose : FunctionoftheWesternBlot istodetectthepresenceorabsenceofproteins inasample,wheretheproteinwastransferredtoaspecialmembranewhichfurtherstainedinordertovisualized.Result :

    1. ResultfromWesternBlotprocess.

  • Conclusions :1. Westernblotprocedure consistsof samplepreparation,gelelectrophoresis, transfer

    fromgeltomembrane,andimmunestainingoftheblot.2. Principlesusedinthewesternblotistheprincipleofantigenbindingcomplexatibodi.

    Protein inNCwe thinkofasantigen.Primaryantibody isanantibody that canbindspecifically toantigens inNC.Tobeable tosee thebondingof theantigenantibodycomplexthenwegivecolortotheantigenantibody.

    3. Ofthisprocesscanbeseen intheformofviralproteinantigens inasamplewhich isdetermined by comparing the band positions obtainedwithMarker position besidehim.

    ThankYouVeryMuch.

  • ReportofActivitySensei :Prof.SatoshiTeraoAssistant :MiyakoKiso(PhD.Student)NamaStudent:BaharuddinBadi,S.SiJapaneseLanguageandCulturePurposes:

    1. ToKnowandunderstandrulesofJapaneseLanguage.2. ToknowandunderstandaboutJapaneseCulture.

    Result:

    1. JapaneseLanguageandCulturesessionclass.Descriptions:

    A. Whileteachingandlearningprocessintheclassroom.B. ThemainhandbookwhichuseduringinJapaneseLanguageclass.

    2. TriptoaroundofKobeUniversityinRokkohighlandarea.

    Descriptions:

    A. Meet and acquaintedwith Indonesian studentswhich are studying atKobeUniversity.

    B. The hospital in Rokko campus has functions to support the health everystudents and teachersduring school atKobeUniversity and for realize thateveryyear,everystudentsmustcometohereforcheckuphis/herconditions.

  • 3. Excursion toCenterofKobeMuseum for learn the livesof Japanesecitizens fromagetoage.

    4. WhilewasmeetingwithMr.TakayoshiKunoM.DPh.D.andteam fromTheCenterforEducationofGlobalLeadersforASEANandStudentsIndonesianJapanExchangeProjectandsomestudentsfromMedicalFacultyofKobeUniversity.

    Conlusions :

    1. In Japanese Language thereare threeof characterswhich composea sentences.The characters areused isKanji,Hiragana andKatakana.Kanji characters is camefromChineandHiragana isoriginwords from JapanbutKatana charactersuse todetermineasomethingofforeign.

  • 2. Thedailyof Japanesepeopleshowahabitofdisciplinesandalways focusto finishhis/herjobs.

    3. TheJapancountryhasbecomeagreatcountryinglobalmarketbecausetheyalwaysdoinginnovationtothemproducts.

    4. ThegovernmentofJapanhaspreventedtrapjamwithuseselectricaltrains,andonthestreetsthegovernmenthasimplementedstrictregulationforcontrolofquantityofvehiclewhichwillreducepollutionsontheair.

    5. The country of Japan very susceptible for earthquake because the Japan islandstandingontopoftheoceanicplateandcontinentalplatewhichnottoostabile,soevery peoples which stay in Japan must know how to indication event theearthquakeandknowofthewarningsystem.

    ThankYouVeryMuch.