TheAscorbicAcid(L-Ascorbate)assaykitisforthespecificmeasurementandanalysisofL-ascorbicacidinbeverages,meat,flour,dairyandvegetableproducts.
Grapeandwineanalysis:Oenologiststoexploitadvancedtestkits.
Charnock,S.C.&McCleary,B.V.(2005).RevuedesEnology,117,1-5.
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Itiswithoutdoubtthattestingplaysapivotalrolethroughoutthewholeofthevinificationprocess.Toproducethebestposs
IBLequalitywineandtominimiseprocessproblemssuchas“stuck”fermentationortroublesomeinfections,itisnowrecognisedthatifpossibletestingshouldbeginpriortoharvestingofthegrapesandcontinuethroughtobottling.Tr
ADItionalmethodsofwineanalysisareoftenexpensive,timeconsuming,requireeitherelaborateequipmentorspecialistexpertiseandfrequentlylackaccuracy.However,enzymaticbio-analysisenablestheaccuratemeasurementofthevastmajorityofanalytesofinteresttothewinemaker,usingjustonepieceofapparatus,thespectrophotometer(
seepreviousissueNo.116foradetailedtechnicalreview).Grapejuiceandwineareamenabletoenzymatictestingasbeingliquidstheyarehomogenous,easytomanipulate,andcangenerallybeanalysedwithoutanysamplepreparation.
Megazyme“advanced”winetestkitsgeneralcharacteristicsandvalidation.
Charnock,S.J.,McCleary,B.V.,Daverede,C.&Gallant,P.(2006).ReveuedesOenologues,120,1-5.
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ManyoftheenzymatictestkitsareofficialmethodsofprestigiousorganisationssuchastheAssociationofOfficialAnalyticalChemicals(AOAC)andtheAmericanAssociationofCerealChemists(AACC)inresponsetotheinterestfromoenologists.Megazymedecidedtouseitslonghistoryofenzymaticbio-analysistomakeasignificantcontributiontothewineindustry,bythedevelopmentofarangeofadvancedenzymatictestkits.Thistaskhasnowbeensuccessfullycompletedthroughthestrategicandcomprehensiveprocessofidentifyinglimitationsofexistingenzymaticbio-analysistestkitswheretheyoccurred,andthenusingadvancedtechniques,suchasmolecular
BIOLOGy(
photo1),torapidlyovercomethem.Noveltestkitshavealsobeendevelopedforanalytesofemerginginteresttotheoenologist,suchasyeastavailablenitrogen(
YAN;seepages2-3ofissue117article),orwherepreviouslyenzymesweresimplyeithernotavailable,orweretooexpensivetoemploy,suchasforD-mannitolanalysis.
BioactivecompoundsfromendemicplantsofSouthwestPortugal:Inhibitionofacetylcholinesteraseandradicalscavengingactivities.
Tavares,L.,Fortalezas,S.,Tyagi,M.,Barata,D.,Serra,A.T.,MartinsDuarte,C.M.M.,Duarte,R.O.,Felicano,R.P.,Bronze,M.R.,Espírito-Santo,M.D.,Ferreira,R.B.&Santos,C.N.(2012).PharmaceuticalBiology,50(2),239-246.
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Context:Naturalproductsarereportedtohavesubstantialneuroprotectiveactivityduetotheirradicalscavengingcapacity,andalsoacetylcholinesterase(AChE)inhibitorycapacity,bothactivitiesimportantinneurodegeneration.Objective:Theundesirablesideeffectsofcompoundsinpharmacologicalusemakeitimportanttoidentifynaturalneuroprotectivemolecules.ThisworkassessesthepotentialoffiveendemicPortugueseplantsassourcesofneuroprotectivecompounds.Materialsandmethods:AntioxidantcapacityforperoxylradicalwasdeterminedbyOxygenRadicalAbsorbanceCapacitymethodandforhydroxylbyElectronParamagneticResonance,aswellasAChEinhibitorycapacityoftheplanthydroethanolicextracts.ThemoleculesresponsibleforthesevaluablepropertieswerealsotentativelyidentifiedbyHPLC.Resultsanddiscussion:ArmeriarouyanaandThymuscapitellatuspresentedsomeofthehighestphenoliccontents(76.60±7.19and12.82±0.24mgGAEg-1dw,respectively)andantioxidantcapacities(592±116and449±57μmolTEg-1dw,respectively).Theflavonoidswereidentifiedasthephytomoleculesrelatedtotheantioxidantcapacityoftheseplantextracts;inthecaseofA.rouyana,L-ascorbicacidalsomadeanimportantcontribution(3.27±0.26mgg-1dw).PlantextractsclearlydemonstratedeffectiveAChEinhibitoryactivity(480±98and490±46μgmL-1,respectively),thatcouldbeassociatedtopolyphenols.Conclusions:TheextractsofA.rouyanaandT.capitellatusandtheiractivecomponents,especiallypolyphenols,demonstrateinterestingneuroprotectivepotential.They,therefore,deservefurtherstudyastheirphytomoleculesarepromisingsourcesofeithernaturalneuroprotectiveproductsand/ornovelleadcompounds.
Effectof1-MCPonqualityandantioxidantcapacityofinvitrodigestsfrom‘Sunrise’applesstoredatdifferenttemperatures.
Qiu,S.,Lu,C.,Li,X.&Toivonen,P.(2009).FoodResearchInternational,42(3),337-342.
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Theantioxidantcapacitiesofphenolicandnon-phenolicfractionsforinvitrodigestatesfrom‘Sunrise’applewereassessedafterpostharvestapplicationof1-methylcyclopropene(1-MCP),aripeninginhibitor,andthreeweeksstorageat5,13,15,18and22°C.Aninvitrodigestionsystemwasusedtogeneratethesolublebioaccessabledigestatewhichwasthenfractionatedintophenolicandnon-phenolicfractions.ThetwofractionswereassayedforFolin-CiocalteuReaction(FCR)reducingcapacityandperoxylradicalscavengingcapacity.Qualityretentionofthefruitwasassessedbymeasuringinternalethyleneconcentration,firmnessandtitratableacidity.Treatmentwith1-MCPinhibitedinternalethyleneconcentrationandbettermaintainedthefirmnessandtitratableacidityof‘Sunrise’summerapplesascomparedwithuntreatedcontrolapplesatstoragetemperaturesof15°Candabove.TheFCRreducingcapacityofthephenolicfractionoftheinvitro,simulatedgastrointestinaldigestatesshowedsimilarresponseasdidthequalitymeasures,withsignificantlyhigheractivityinthe1-MCPtreatedfruitathigherstoragetemperatures.However,noconsistentdifferenceswerefoundbetween1-MCPandcontroltreatmentsfortheFCRreducingcapacityofthenon-phenolicfractionorfortheperoxylradicalscavengingcapacityofeitherfraction.Thenon-phenolicfractionsconsistentlyhadhigherlevelsofbothtypesofantioxidantcapacities.Treatmentandstorageof‘Sunrise’applesatelevatedtemperatures(>13°C)resultedinimprovedfruitqualityandretentionofreducingcapacityinsimulatedgastrointestinaldigestates.
Influenceofstarterculturesontheantioxidantactivityofkombuchabeverage.
Malbaša,R.V.,Lončar,E.S.,Vitas,J.S.&Čanadanović-Brunet,J.M.(2011).FoodChemistry,127(4),1727-1731.
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Thispaperinvestigatestheinfluenceofstartercultures,obtainedfromkombuchaisolates,ontheantioxidantactivityofkombuchabeverages.Threestartercultureswereusedasfollows:(1)mixedcultureofaceticbacteriaandZygosaccharomycessp.(SC1);(2)mixedcultureofaceticbacteriaandSaccharomycescerevisiae(SC2);aswellas(3)nativelocalkombucha.Thestartercultureswereaddedtoblackandgreenteasweetenedwith7%ofsucrose.Fermentationwascarriedoutat28°Cfor10days.AntioxidantactivitytohydroxylandDPPHradicalswasmonitored.KombuchabeverageonblackteahasshownthehighestantioxidantactivitytobothtypesofradicalswithstarterSC1,whilethegreenteabeveragehasshownthehighestactivitywithnativekombucha.Themainreasonforthedifferentantioxidantactivities,besideteacomposition,wasascribedtodifferingproductionofbothvitaminCandtotalorganicacidsintheinvestigatedsystems.
AntioxidantcapacityofMacaronesiantraditionalmedicinalplants.
Tavares,L.,Carrilho,D.,Tyagi,M.,Barata,D.,Serra,A.T.,Duarte,C.M.M.,Duarte,R.O.,Feliciano,R.P.,Bronze,M.R.,Chicau,P.,Espírito-Santo,M.D.,Ferreira,R.B.&DosSantos,C.N.(2010).Molecules,15(4),2576-2592.
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Theuseofmanytraditionalmedicinalplantsisoftenhamperedbytheabsenceofaproperbiochemicalcharacterization,essentialtoidentifythebioactivecompoundspresent.TheleavesfromfivespeciesendemictotheMacaronesianislandswithrecognizedethnobotanicalapplicationswereanalysed:
Apolloniasbarbujana(Cav.)
Bornm.,
Ocoteafoetens(Ainton)Baill,
Prunusazorica(Mouill.)Rivas-Mart.,Lousã,Fern.Prieto,E.Días,J.C.Costa&C.Aguiar,
RumexmaderensisLoweand
PlantagoarborescensPoir.subsp.
maderensis(Dcne.)A.Hans.etKunk.Sinceoxidativestressisacommonfeatureofmostdiseasestraditionallytreatedbytheseplants,itisimportanttoassesstheirantioxidantcapacityanddeterminethemoleculesresponsibleforthiscapacity.Inthisstudy,theantioxidantcapacityoftheseplantsagainsttwoofthemostimportantreactivespeciesinhumanbody(hydroxylandperoxylradicals)wasdetermined.Totracetheantioxidantorigintotalphenolandflavonoidcontentsaswellasthepolyphenolicprofileandtheamountoftraceelementsweredetermined.Therewasawidevariationamongthespeciesanalysedinwhatconcernstheirtotalleafphenolandflavonoidcontents.FromtheHighPerformanceLiquidChromatography(HPLC)elect
Rochemicallydetectedpeaksitwaspossibletoattributetoflavonoidstheantioxidantcapacitydetectedin
A.barbujana,
O.foetens,
R.maderensisand
P.azoricaextracts.Thesepotentialreactiveflavonoidswereidentifiedfor
A.barbujana,
R.maderensisand
P.azorica.For
R.maderensisahighcontent(7mgg
-1dryweight)ofL-ascorbicacid,analreadydescribedantioxidantphytomolecule,wasfound.Ahighcontentinselenomethionine(414.35μgg
-1dryweight)wasobtainedfor
P.arborescenssubsp.
maderensisextract.Thisselenocompoundisalreadydescribedasahydroxylradicalscavengerisreportedinthisworkasalsopossessingperoxylradicalscavengingcapacity.Thisworkisagoodillustrationofdifferentphytomolecules(flavonoids,organicacidsandselenocompounds),presentsinleavesofthefivetraditionalmedicinalplantsendemictoMacaronesia,allexhibitingantioxidantproperties.
Nutritionalandsensoryqualityduringrefrigeratedstorageoffresh-cutmints(MenthaxpiperitaandM.spicata).
Curutchet,A.,Dellacassa,E.,Ringuelet,J.A.,Chaves,A.R.&Viña,S.Z.(2014).FoodChemistry,143,231-238.
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Theeffectofstoragetimeonqualityattributesofrefrigeratedfresh-cutmints(Mentha×piperitaandM.spicata)wasstudied.Atmospherecomposition,respiratoryactivity,weightloss,surfacecolour,totalchlorophyll,carotenoids,browningpotential,totalphenols,flavonoids,radical-scavengingactivity,ascorbicacidandessentialoilyieldandcompositionwereanalysed.Respiratoryactivityofpeppermintandspearmintsamplesdiminishedmoderately(42%and28%,respectively)after21daysat0°C.Aslightmodificationoftheinternalatmospherewasachieved.Surfacecolour,chlorophyll,carotenoidandantioxidantcompoundsremainedalmostconstant.Theyieldofessentialoildidnotchangeoritshowedanapparentincreaseafter21daysat0°C,dependingonplantgrowthstage.Thecharacteristicflavourcomponentsofpeppermint(menthoneandmenthol)increased,whilethecontentsofthemainconstituentsofspearmintessentialoilshowedminorvariationsafterstorage.Theconditionsassayedforpackagingandstoringfresh-cutmintswereadequatetoachievearelativelylongshelflifeandtheyretainedtheirantioxidantproperties.
Ahigh‐grainproteincontentlocusonbarley(Hordeumvulgare)chromosome6isassociatedwithincreasedflagleafproteolysisandnitrogenremobilization.
Jukanti,A.K.&Fischer,A.M.(2008).PhysiologiaPlantarum,132(4),426-439.
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Leafsenescenceandnitrogenremobilizationfromsenescingtissuesaretwoimportantfactorsdetermininggrainproteincontent(GPC)incereals.Wecomparednear-isogenicbarley(
HordeumvulgareL.)germplasmvaryingintheallelicstateofamajorGPCquantitativetraitlocusonchromosome6,delineatedbymolecular
MarkersHVM74andABG458andexplainingapproximately46%ofthevari
ABIlityinthistrait.HighGPCwasconsistentlyassociatedwithearlierwhole-plantsenescence.SDS–PAGEandimmunoblotanalysisofflagleafproteinsindicatedearlierleafprotein[includingribulose-1,5-bisphosphatecarboxylase/oxygenase(Rubisco)]degradationinhigh-GPCgermplasm.Thiswasaccompaniedbyenhancedavailabilityofammoniumandglutamineindevelopingkernels,suggestingincreasedphloemretranslocationofnitrogen.Basedonpreviousmicroarrayanalysis,weperformedadetailedexpressionstudyofsixleafgenes,tentativelyinvolvedinplastidialproteolysis,vacuolarproteolysis,intermediaryNmetabolismandNtransport.Allofthesewereupregulatedinhigh-GPCbarley,mostlyaround21to28dayspastanthesis,priortooraroundthetimedemonstratingmaximaldifferencesinleafprotein(includingRubisco)levels.Therefore,thesegenesrepresentpotentialtargetstomanipulategrainproteinaccumulation.Itappearslikelythattheirfunctionalanalysiswillenhanceourunderstandingofwhole-plantNrecycling.Additionally,earlierleaf(photosynthetic)proteindegradationmayleadtoreducedNcarbonassimilationinhigh-GPCgermplasm,explainingpaststudiesdemonstratinganegativecorrelationbetweenGPCandyield.
AmutationinGDP-mannosepyrophosphorylasecausesconditionalhypersensitivitytoammonium,resultinginArabidopsisrootgrowthinhibition,alteredammoniummetabolism,andhormonehomeostasis.
Barth,C.,Gouzd,Z.A.,Steele,H.P.,&Imperio,R.M.(2010).JournalofExperimentalBotany,61(2),379-394.
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Ascorbicacid(AA)isanantioxidantfulfillingamultitudeofcellularfunctions.Givenitspivotalroleinmaintainingtherateofcellgrowthanddivisioninthequiescentcentreoftheroot,itwashypothesizedthattheAA-deficient
Arabidopsisthalianamutants
vtc1-1,
vtc2-1,
vtc3-1,and
vtc4-1havealteredrootgrowth.Totestthishypothesis,rootdevelopmentwasstudiedinthewildtypeand
vtcmutantsgrownonMurashigeandSkoogmedium.Itwasdiscovered,however,thatonlythe
vtc1-1mutanthasstronglyretardedrootgrowth,whiletheother
vtcmutantsexhibitawild-typerootphenotype.Itisdemonstratedthattheshort-rootphenotypein
vtc1-1isindependentofAAdeficiencyandoxidativestress.Instead,
vtc1-1isconditionallyhypersensitivetoammonium(NH
4+).ToprovidenewinsightsintothemechanismofNH
4+sensitivityin
vtc1-1,rootdevelopment,NH
4+content,glutaminesynthetase(GS)activity,glutamatedehydrogenaseactivity,andglutaminecontentwereassessedinwild-typeand
vtc1-1mutantplantsgrowninthepresenceandabsenceofhighNH
4+andtheGSinhibitorMSO.Since
VTC1encodesaGDP-mannosepyrophosphorylase,anenzymegeneratingGDP-mannoseforAAbiosynthesisandproteinN-glycosylation,itwasalsotestedwhetherproteinN-glycosylationisaffectedin
vtc1-1.Fur
Thermore,sincerootdevelopmentrequirestheactionofavarietyofhormones,itwasinvestigatedwhetherhormonehomeostasisislinkedtoNH
4+sensitivityin
vtc1-1.OurdatasuggestthatNH
4+hypersensitivityin
vtc1-1iscausedbydisturbedN-glycosylationandthatitisassociatedwithauxinandethylenehomeostasisand/ornitricoxidesignalling.
ReductionofacrylamideformationbyvanadiumsaltinpotatoFrenchfriesandchips.
Kalita,D.&Jayanty,S.S.(2013).FoodChemistry,138(1),644-649.
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Theeffectsofvanadylsulphateontheformationofacrylamidehavebeenstudiedinfriedpotatoproducts,suchasFrenchfriesandchips.Acrylamideformationwasinhibitedby30.3%,53.3%and89.3%whentheslicedpotatostripsweresoakedin0.001,0.01and0.1Mvanadylsulphate(VOSO4)solutions,respectively,for60minbeforefrying.Moreover,57.7%,71.4%and92.5%inhibitionofacrylamideformationwasobservedwhenchipsweresoakedintherespectivevanadylsulphatesolutionbeforefrying.Inaseparatemodelreaction,asolutioncontaininganequimolarconcentrationofL-asparagineandD-glucoseshowedasignificantinhibitionofacrylamideformationwhenheatedat150°Cfor30mininthepresenceofvanadylsulphate(VOSO4).TheresultsindicatethatthebindingofVO2+toasparagineandthedecreaseinthepHofthepotatosamplesresultedinasignificantreductionofacrylamideformationinfriedpotatoproducts.
Roleofpolyphenolsinacrylamideformationinthefriedproductsofpotatotuberswithcoloredflesh.
Kalita,D.,Holm,D.G.&Jayanty,S.S.(2013).FoodResearchInternational,54(1),753-759.
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Thelevelsofasparagine,reducingsugarsandtotalphenolicsinsomepotatocultivarsandadvancedselectionswithdistinctivefleshcolor(white,yellow,redandpurple)andtheirpotentialinacrylamideformationinFrenchfriesandpotatochipshavebeeninvestigatedinthisstudy.Therangeofasparagineandreducingsugarswere1.8–9.0and1.35–11.7mg/grespectively.Therewasnosignificantcorrelationofasparagineandreducingsugarswithfleshcolorofthepotatotubers.Tuberswithredandpurplefleshhadhigherlevelsoftotalphenolicsthanthewhiteandyellowones.TheamountofacrylamideformedinFrenchfriesandchipswererangedfrom128.1to1651.3μg/kgandfrom2104.3to2978.5μg/kgrespectively.Thelevelsofasparagineandreducingsugarswerepositivelycorrelatedwithacrylamideformationwhiletotalphenolicsandchlorogenicacidcorrelatednegatively.
cMycincreasescellnumberthroughuncouplingofcelldivisionfromcellsizeinCHOcells.
Kuystermans,D.&Al-Rubeai,M.(2009).BMCBiotechnology,9(76).
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Background:Overthepastdecades,theincreaseinmaximalcellnumbersfortheproductionofmammalianderivedbiologicshasbeeninalargepartduetothedevelopmentofoptimalfeedingstrategies.Engineeringofthecelllineisoneofprobableapproachesforincreasingcellnumbersinbioreactor.Results:Wehavedemonstratedthattheover-expressionofthec-mycgeneinimmortalisedCHOcellscanincreaseproliferationrateandmaximalcelldensityinbatchculturecomparedtothecontrol.ThechangeswereattributedtoarapidtransitionintoS-phasefromashorteneddurationofG1phaseandtotheuncouplingofcellsizefromcellproliferation.Toachievethe>70%increaseinmaximalcelldensitywithoutadditionalsupplyofnutrientsthecellsunderwentanoverallreductionof14%insizeaswellasasignificantdecreaseinglucoseandaminoacidconsumptionrate.Consequently,thetotalbiomassaccumulationdidnotshowasignificantchangefromthecontrol.TheamountofhSEAP-hFcactivityoftheoverexpressingc-myccelllinewasfoundtobewithin0.7%ofthecontrol.Conclusion:ItisshownthatthemanipulationofcellcyclekineticsandindirectlycellmetabolismgiveshighercelldensitiesinCHObatchcultures.Theunalteredapoptoticratesupportedthepropositionthattheincreaseincellnumberwasaresultofenhancecellcyclekineticsandcellularmetabolismratherthanincreasingviability.ProductionofhSEAP-hFcfromaconstitutivec-mycover-expressingcelllinedidnotincreasewiththeincreaseincellnumber.