Statistics Question

statistics writing question and need the explanation and answer to help me learn.

Follow the documents I have posted, please. All information is within the documents.
https://www.proquest.com/docview/2347546923?accountid=12085&forcedol=true&forcedol=true
Requirements: Refer to documents
Research Exploration Assignment Instructions
Overview
There are 4 Research Exploration Assignments in this course, and they are designed to help you explore major concepts and questions related to human development and to enhance your skill in reading and interacting with scholarly literature. You will gain knowledge about prenatal development and will also develop skills that will be helpful as you research other topics in the future.
Instructions
There are two parts to this assignment:
First, you will access the article within the resources section of this assignment and download a PDF copy of the assigned article. Using the highlighting and commenting tools provided within the Adobe PDF program, you will identify the following elements within the article. Be sure to number each element of the article according to the numbered list adopted below:
Title
Author(s)
Journal name
Date of publication
Purpose statement.
Key finding/conclusion (highlight this in the article, not in the Abstract).
In a separate document, respond to the following questions/prompts providing 3-5 complete sentences for each question/prompt (keep in mind that question 4 is an exception to the 3-5 sentence requirement).
Why did the authors write this article? (No direct quotes, but cite your source).
What is the main idea of the article? (No direct quotes, but cite your source).
To whom would this information be relevant and why? (No direct quotes, no citation required).
Include a brief quotation from the article that you found particularly interesting. Be sure to enclose this quote in quotation marks and include a properly-formatted citation.
What is a fact/concept from this article you might share with someone you know? Write this in your own words, but include an APA-formatted citation to identify the source.
Identify a Biblical concept that supports the ideas explored in this article. Explain (in 3-5 sentences) why you believe there is a connection. Include a properly formatted citation for the scripture reference.
Assignment Requirements
Students will upload the edited copy of the assigned article (PDF).
For the separate document (Q & A) please include the following:
Use the current APA format on the APA-formatted title page.
Response to the questions/prompts divided by level-one headings (one heading for each question/prompt). A Research Exploration Assignment Sample Paper is provided in the Research Exploration Resources of each assignment to use as an example/template.
An APA-formatted reference page.
Note: Your assignment will be checked for originality via the Turnitin plagiarism tool.

Research Exploration Assignment
Chris T. Sample
School of Behavioral Sciences, Liberty University
PSYC 210: Developmental Psychology
Professor Smith
March 3, 2020
Why Was This Article Written
Based on the information in this article, write 3-5 sentences (in your own words) explaining why the authors wrote this article. Remember to include an APA-formatted citation to identify your source of information. For example, the author attempted to explain why she believes that science supports the concept that life begins at conception. In addition to providing information about why life begins at conception, ideas supporting the idea that life does not begin at conception are refuted (Irving, 1999).
What is the Main Idea of this Article
Based on the information in this article, write 3-5 sentences (in your own words) identifying the main idea of this article. Remember to include an APA-formatted citation to identify your source of information.
To Whom Would this Article be Relevant
Based on the information in this article, write 3-5 sentences (in your own words) identifying at least one audience (person, groups of people, etc.) that would benefit from reading this article. Explain your choice. No citation is required for this section.
Interesting Quote from this Article
Select a brief quotation from the article that you found particularly interesting. Be sure to copy the quote word for word, enclose this quote in quotation marks, and include a properly-formatted citation. Remember when citing a direct quote you should include the authors’ names, the date of publication, and a page number. Example (Irving, 1999, p. 345).
Shareable Content from this Article
Include a fact/concept from this article you might share with someone you know. Write this in your own words, but include an APA-formatted citation to identify the source.
Biblical Concept Related to the Article
Identify a Biblical concept that supports the ideas explored in this article. Explain why you believe there is a connection. Include a properly formatted citation for the scripture reference. Citation example: The Bible states, “For God so loved the world, that he gave his only Son, that whoever believes in him should not perish but have eternal life” (English Standard Bible, 2001/2016, John 3:16).

References
English Standard Bible. (2016). (Original work published 2001)
Irving, D. N. (1999). When do human beings begin? Scientific myths and scientific facts. International Journal of Sociology and Social Policy, 19(3/4), 22-46.

ArticleSleep,LittleBaby:TheCalmingEectsofPrenatalSpeechExposureonNewborns’SleepandHeartrateAdelheidLang1,2,*,RenatadelGiudice1,2,3,4andManuelSchabus1,21DepartmentofPsychology,CentreforCognitiveNeuroscience(CCNS),UniversityofSalzburg,5020Salzburg,Austria;renata.delgiudice@asst-santipaolocarlo.it(R.d.G.);manuel.schabus@sbg.ac.at(M.S.)2LaboratoryforSleep,CognitionandConsciousnessResearch,UniversityofSalzburg,5020Salzburg,Austria3DepartmentofMentalHealth,UniversityofMilan,20142Milan,Italy4SanPaoloUniversityHospital,ASSTSantiPaoloeCarlo,20142Milan,Italy*Correspondence:adelheid.lang@sbg.ac.at;Tel.:+43-662-8044-5113Received:26June2020;Accepted:30July2020;Published:2August2020Abstract:Inapilotstudy,34fetuseswerestimulateddailywithamaternalspokennurseryrhymefromweek34ofgestationonwardandre-exposedtwoandfiveweeksafterbirthtothisfamiliar,aswellastoanunfamiliarrhyme,bothspokenwiththematernalandanunfamiliarfemalevoice.Duringauditorystimulation,newbornswerecontinuouslymonitoredwithpolysomnographyusingvideo-monitoredhdEEG.Afterward,changesinsleep–wake-stateproportionsduringfamiliarandunfamiliarvoicestimulationwereanalyzed.Ourpreliminaryresultsdemonstrateageneralcalmingeectofauditorystimulationexclusivelyininfantswhowereprenatally“familiarized”withaspokennurseryrhyme,asevidencedbylesswakingstates,moretimespentinquiet(deep)sleep,andlowerheartrates.Astimulationnaïvegroup,ontheotherhand,demonstratednosucheects.Stimulus-specificeectsrelatedtothefamiliarityoftheprenatallyreplayedvoiceorrhymewerenotevidentinnewborns.Together,theseresultssuggest“fetallearning”atabasiclevelandpointtoafamiliarizationwithauditorystimulipriortobirth,whichisevidentinthefirstweeksoflifeinbehavioralstatesandheartratephysiologyofthenewborn.Keywords:perinatalmemory;fetallearning;quietsleep;activesleep;fetus;newborn;maternalvoice;EEG1.IntroductionInfants’sleepinghabitsareaconstantconcernforparents,includingthequestionofwhysomeinfantscandriftotosleepsoeasilyandstillsleeponeveninunfamiliarornoisyenvironments,whereasotherssimplycannotinthepresenceoftheslightestauditorycues.Besidesgeneticandbiologicalreasons,oneplausibleanswercouldbeinfants’prenatalexperiences,oftenalsoreferredtoas“fetalprogramming.”Theindividualvariancethatfetusesexperiencewhenitcomestoauditoryexposuredependsontheeverydaymaternalenvironmentandisstronglyinfluencedbyfactorssuchasthemother’sdailyroutinesandsocialhabitsorexposuretoenvironmentalnoise.Asapilotstudyforaplannedlongitudinalexaminationregardingtheperinatalimpactofstressoninfants’development,weexploredthequestionofprenatalinfluencesonnewborns’sleepandphysiology.Indetail,wewereinterestedinwhetherrepeatedprenatalauditorystimulationwithatapedmaternalspokennurseryrhymeleadstofamiliarizationinfetusesandthereforelesssleepdisruptionsinauditoryre-exposednewborns.Furthermore,wewereinterestedinthespecificeectsofstimulusfamiliarity(maternalvs.unfamiliarvoice;prenatallyfamiliarizedvs.unfamiliarnurseryrhyme)oninfant’ssleep–wake-statesandphysiology.BrainSci.2020,10,511;doi:10.3390/brainsci10080511www.mdpi.com/journal/brainsci
BrainSci.2020,10,5112of11Whatisalreadyknownaboutsleepduringtheperinatalperiod?Third-trimesterfetusesandnewbornsareknowntospendthevastmajorityoftheirtimesleeping[1,2],whichiscompatiblewiththeparticularrelevanceofsleepforbrainandbodydevelopment.Asthecircadianrhythmisnotfullyestablishedeveninhealthyfull-termnewborns,newbornsareknowntoshowanirregularyetextensivepatternofsleep(upto18h)andwakefulnessduringdayandnight,withsleepoftenonlyinterruptedbyfeedingperiods[3].Newborn’ssleepisdividedintothreestages[1]:Quietsleep(QS;similartoadultnon-rapid-eye-movementsleep)withnoorminimalbodyandeyemovements,regularrespirationandheartrate;activesleep(AS;similartoadultrapid-eye-movementsleep)withirregularrespiration,facialgrimaces,smiling,sucking,muscletwitches,andgrosslimbmovements;aswellastransitionalsleep(TS),whichusuallymarkstransitionsfromwake(W)toASandviceversa.Incontrasttoolderinfants,thenewbornsleepcycleusuallystartswithASandischaracterizedbyanequalproportionofASandQS[3],withASsteadilydecreasingforthesakeofmoreQSoverthefirstweeksoflife[4].Sleepandsleeppatternsinnewbornsareinfluencedbyseveralfactors.Besidescentralnervoussystemmaturation[5],thedepthofsleepisreportedtobeinfluencedbythesleepingposition(QSisenhancedinsupinevs.proneposition)[6],aswellascaregivingpractices(suchastheamountofmaternalskin-to-skincontact)[7].Additionally,thereareafewstudiesinvestigatingchangesinnewborn’ssleeppatternsinresponsetoenvironmentalnoise.Forexample,QSisfoundtobeincreasedinpretermsafterreducingambientnoise[8,9]andsleepdisruptionsincreasetounfamiliarsoundsinpreterminfants,evenifsoundpeakswereonlymoderatelyabovebackgroundnoiselevels[10].Ontheotherhand,Filippaetal.[11]foundthatlisteningtothematernalvoice—probablythemostsalientstimulusfornewborns—leadstoarousingshiftsfromsleeptowakewhennewbornswereexposedtomaternalspeechbutnotwhentheywereexposedtothesingingvoiceofthemother[11].Itisnoteworthythat,inpaststudies,thematernalvoiceissometimesinterpretedasarousing[12],whereasinotherstudies,itisinterpretedasrathercalming[13,14],asoftenevidencedbyheartratechanges.Onlyafewofthestudiespublishedactuallyreportthebehavioralstate(wake,AS,QS)ofthebabiesatstimulationonset.Eventually,thedierentialbehavioralreactions(calmingvs.arousing)foundinresponsetoauditorystimulationmaythusberelatedtosuchstateeects.Ourstudynowfocusesexplicitlyonsleep-wake-statechangesto(prenatallypresentedornotpresented)auditorystimuli.Giventheliterature,itiswelldocumentedthatevennewbornsstayreceptibletospecificincominginformationevenatovertunconsciousand“oine”periodsofsleep.Indetail,ithasbeenreportedthatsleepingnewbornsareabletodistinguishbetweenvariousauditorystimuli,asdemonstratedforspeechvs.music[15],standardvs.deviantvowels[16,17],andforwardvs.backwardvoices[18].Furthermore,sleepinginfantsarereportedtodetectprosodicspeechcues[19],discriminateangerandfearinvoices[20],andevenlearnduringsleep,asevidencedbyincreasedeyemovementresponsestoatoneearlierpairedwithanairpu[21,22].Also,sleepingnewborns(agedtwodaystosixweeks)arereportedtoshowheartratede-oraccelerationstovariousauditorystimuli[11,23,24].However,thereisaninterestingindividualvariationintheintensitylevelneededtoevokearesponseinnewbornsduringdierentbehavioralstates(foranoverviewsee[24]).Additionally,ithasbeendemonstratedthatfetuses’[25,26]andnewborns’[24,27,28]responsestosoundsare,amongstothers,dependentonprevioussoundexposure.Surprisingly,theseresponsestofamiliarstimuliaresometimesreportedtobestronger[24,28],indicatinganorientingresponse,andsometimesweaker[25–27],indicatinghabituation,ascomparedtonewandunfamiliarstimuli.Thesestimulusfamiliaritystudiesallowtheinvestigationoftheearliestformsofmemory,asitsoriginslieinsomekindoflearningor“familiarization”beforestimulusre-exposure.Evidenceforauditorylearninginfetuses(i.e.,dierentialresponsestofamiliarvs.unfamiliarstimuli)hasbeenreportedfromthethirdtrimesterofgestationonwards.Fetusesareabletolearnmusic[28,29],voices,stories,andnurseryrhymes(foranoverview,seeKisilevsky&Hains[30]),asevidencedbyfetalheartratechangestothesefamiliarstimuli.Furthermore,decreasingstartleresponsesormovementstoarepeatedlypresentedvibroacousticstimulushasbeenshownatthisearlystage[25,26],aswellas
BrainSci.2020,10,5113of11habituationeectstorepeatedlypresentedtonesonabrain-levelusingfetalMagnetoencephalography(MEG)[31].Additionally,behavioralstatetransitionshavebeenreportedtobecomemoreprevalentwhenfetusesarere-exposedtofamiliarmusic[28].Suchkindsoffetalmemorieshavebeenshowntopersistintotheneonatalperiodandthefirstmonthsoflife.Themoststudiedstimulusinthiscontextisagainthematernalvoice.Afterbirth,newbornsarereportedtoreactquitedierentlytothematernalvoice,asevidencedbychangesinsuckingbehaviorofpacifiers[32,33],decreasingamountofmovements[34],orfewerbehavioralsignsofpainduringbloodsamplingwhenthematernalvoicehasbeenreplayedatthesametime[35].Besidesthematernalvoice,thereisalsoevidencethatprenatallypresentedmusicisrecognizedatbirth,asevidencedbyheartrate(HR)decreases[36]andsleeptowaketransitionsinnewborns[28].Insummary,thereisgoodevidenceforprenatallearningandtheretentionofprenatalmemorytracesuptothepostnatalperiod.Inthepresentstudy,wethereforeexaminedchangesinbehavioralstates(W,QS,AS)toauditorystimulationinprenatallyfamiliarized(EG)orstimulationnaïve(CG)newborns.Forthatpurpose,wecontinuouslymonitorednewbornswithpolysomnography(i.e.,usingvideo-monitoredhighdensityelectroencephalography;hdEEG),includingelectromyogram,electrooculogram,electrocardiographyandvideography)atweektwoandweekfiveafterbirthintheirhomeenvironmentwhentheywereexposedtoprenatallypresented“familiar”ornew“unfamiliar”nurseryrhymesineitherthevoiceofthemotheroranunfamiliarfemalevoice.Despitemixedearlierfindings,wehypothesizedincreasedwakingduringstimulationwiththeunfamiliarrhymeintheprenatallyfamiliarizedEG.Furthermore,wehypothesizedthatthefamiliar(maternal)voicewillcalmallnewborns(EGandCG)andresultinlesswaking.Additionally,weexpectedingenerallessarousal,indicatedbylowerheartrates,inprenatallystimulatednewborns(EG).2.MaterialsandMethods2.1.ParticipantsForty-fivepregnantwomenwithaminimumageof18yearsandcomplication-freepregnancieswererecruitedfrom2016to2018atmeetingsforparents-to-beoflocalhospitals.Anadditionalsampleoftenparticipantsdroppedoutbeforepostnataltestingduetoprematurebirthorpregnancy-relatedhealthproblems.Fromthe45babiesincludedinthefullstudy,11infants(EG=6;CG=5)hadtobelaterexcludedfromdataanalysisduetouncorrectableartifacts(frommovements,crying,and/orbadelectrodes)inphysiologicalmeasuresandasaconsequenceambiguoussleepstagingscores.Thefinalsample,therefore,includeddatafrom34infants(EG=23;CG=11)withameanageof14.78days(SD=4.29)inthefirstrecordingandameanageof36.65days(SD=4.49)inthesecondrecording.Allinfantswerebornfull-term(>38weeksofgestation,mean=39.45)andhealthy.Mothers-to-bewereonaverage31.47years(SD=4.29)oldandnativeGermanspeakers.Fifty-threeercentweremarried,47%wereunmarriedbutlivingwiththeirpartner,and42%hadauniversitydegree.Participantsgavewritteninformedconsentattheirfirstvisittoourlaboratory.ThepresentedstudywasapprovedbytheethicscommitteeoftheUniversityofSalzburg(EK-GZ12/2013).2.2.MaterialsWeusedtwodierentecologicallyvalidnurseryrhymesforauditorystimulation,whichweredierentinrhythm(calmvs.lively).Bothrhymesweretapedwiththeindividualmaternalvoiceandanunfamiliarfemalevoice(aprofessionalfemalespeaker,identicaloverallrecordings).UsingthefreelyavailablesoftwareAudacity®(iWebMediaLtd.,Birkirkara,Malta[37]),westandardizedthetotallengthofeachrhymetoexactly60sperrhyme.Astwicedaily(morningandevening)prenatalstimulation(withonerhyme)wasplannedtolastfiveminutesperstimulation,aloopwithfiverepetitionswascreatedwiththeoriginallytapedrhyme,resultinginfiverhymerepetitionsperstimulationday.Forstimulationafterbirth,stimuliwererandomlypresentedoverspeakers(60dB)
BrainSci.2020,10,5114of11withalaptopusingthepresentationsoftwarePresentation(NeuroBehavioralSystems,Berkeley,CA,USA[38]).2.3.ExperimentalProcedureTheexperimentalprotocolwasdividedinaprenatalandpostnatalstimulationpart(seeFigure1).Inpreparationfortheprenatalstimulation,mothers-to-bevisitedourlaboratorybeforegestationalweek34andtapedthetwodierentnurseryrhymesmentionedbefore.Theexperimentalgroup(n=23)receivedoneofthetwomaternalspokenrhymes(i.e.,randomlychosenrhymesofdierentrhythm)tapedonaCDandreplayeditoverspeakers(80dB)fromgestationalweek34onuntilbirth,asweexpect20dBdampeningacrossthemothers’bellyaccordingtoGerhardtandAbrams[39].Auditorystimulationtookplacetwiceaday(morningandevening)forfiveminutes(fiverepetitionsofthe60slastingnurseryrhyme)inthehomeenvironmentofmotherandchild.MothersintheEGwereinstructedtositandrelaxinaquietroomandavoidtouchingtheirbellyduringauditorystimulation.Dailystimulationfrequency,stimulationsoundpressurelevel,andmothers’well-beingwereverifiedanddocumentedusingatabletatthemothers’homes.Anadditionalcontrolgroup(n=11)ofmothersdidnotplayanyrhymebeforebirthbutcompletedanidenticalstudyprotocolthereafter.Atabouttwoandfiveweeksafterbirth,mothersandinfantswerevisitedintheirhomeenvironment(exceptforfourmothers,whodecidedtocometothelabforpostnatalrecordings).Afterinstructingtheparticipants,anhdEEGcap[40]wasplacedontheinfant’shead,andacamera,speakers,andlaptopforstimuluspresentationweresetup.Polysomnography(PSG)includinginfants’electrocardiography(ECG),hdEEG,andvideowerecollectedduringbaseline(silence)periodsaswellasduringauditorystimulationwiththetwodierentnurseryrhymes,tapedandspokenbythemother(familiarvoice)andbyaprofessionalfemalespeaker(unfamiliarvoice).Recordingtimewasintotal27min,withthreeminutes(=threerepetitionsoftheone-minuterhyme)foreachofthefourrandomizedpresentedstimuluscondition(2rhymes2voices)andathree-minutebaseline(silence)beforeeach(andafterthelast)stimulus.Withinaboutfiveweeksafterbirth,thesameexperimentalsettingwasrepeated.Theexperimentalsetupafterbirthwasidenticalfortheexperimentalandcontrolgroup.Figure1.Experimentalprocedure.Pregnantwomen(n=34)wereaskedtotapetwodierentnurseryrhymesandwereafterwardrandomlyassignedtotwogroups.Theexperimentalgroup(EG,n=23)replayedonerhyme(80dBoverspeakers;twiceadayforfiveminutes)fromweek34ofgestationuntilbirth(i.e.,upto~42days).Thecontrolgroup(CG,n=11)didnotreplayanyrhyme.Twoandfiveweeksafterbirthinfants’electrocardiography(ECG)andhighdensityelectroencephalography(hdEEG)wererecordedduringbaseline(silence)andauditorystimulationperiodswithbothrhymes,eachpresentedwiththematernalandanunfamiliarfemalevoice.2.4.ElectrophysiologicalDataCollectionForEEGacquisition,a128electrodesGSNHydroCelGeodesicSensorNet(ElectricalGeodesicInc.,Eugene,OR,USA[40]),providedinthreedierentsizes(34–36,36–37,37–38cm),andaNet
BrainSci.2020,10,5115of11Amps400amplifier(ElectricalGeodesicInc.,Eugene,OR,USA[40]),wereused.Electromyogram(EMG)andhorizontalelectrooculography(EOG)werecollectedusingelectrodesfromtheEEGnet,foradditionalverticalEOG,twoadditionalelectrodeswereplacedaboveandbelowtheinfant’slefteye.Twofurtherelectrodeswereplaceddiagonallyabovetheinfant’srightclavicleandleftabdomenforECGmeasurements.EEGandECGdatawererecordedwithasamplingrateof1000Hz.Duringelectrophysiologicaldatacollection,allbabieswerelyinginthearmoftheirmothers,whowereinstructedtokeeptheirbabystillor,ifthebabygetsuneasy,makeonlygentlerockingmovements.2.5.SleepStagingAnalysisSleepstagingwasbasedonsixEEG(F3,F4,C3,C4,O1,O2),bipolarECG/EMG/VEOG,andHEOGchannels(i.e.,usingaclassicalPSGsetup).Every30-sec-epochwasassignedtowake(W),activesleep(AS),quietsleep(QS),transitionalsleep(TS),orartifact-richepochsofmovement(X),accordingtoScholleandFeldmann-Ulrich[41].Simultaneously,videostapedduringdataacquisitionwerealsoanalyzedin30-sec-epochs,accordingtoPrechtlStates1–5[42].Afterward,sleepstagingandPrechtlstateswerecomparedbyasecondrater,andincaseofamismatch(e.g.,ASinPSGvs.wakeinPrechtl),sleepstagingresultswerecorrected.Inthefinalanalysis,onlysubjectswithunambiguoussleepstagingscoreswereincluded,andepochswithtransitionalsleepandmovementartifactswereexcludedfromanalysis.2.6.ECGPreprocessingandAnalysisECGdatawaspreprocessedusingMatlab(TheMathWorks,Inc.,Natick,MA,USA[43])softwarewiththeAnslabProfessional[44]toolbox.AftercorrectingforartifactsandincorrectlydetectedR-peaks,thedatawas—correspondingtosleepstaginganalysis—segmentedin30sepochs,andinfants’meanheartratewascalculatedforeveryepoch.2.7.StatisticalAnalysesStatisticalanalyseswereperformedusingIBMSPSSStatistics24[45]andbasedonrepeatedmeasuresandmixedanalysesofvariance(ANOVA).Significancelevelsweresettop<0.05andeectsizesarereportedaspartialetasquared(p.eta2)andCohen’sd(d).Greenhouse-Geissercorrectionwasutilizedforviolationsofsphericity.Totestwhetherauditorystimulationinfluencesthesleep–wakecycle,thetime(inseconds)spentinW,AS,andQSwascalculatedforeverysubject.Afterward,wecalculatedthemeantimespentinW,ASandQSforfourbaselines(180seach;orsixbehavioralstatevalueswith30sduration)andfourstimulationphases(also180seach).Theeectofvoicefamiliarityonsleep–wakestateswascomputedbyfurtherdividingpreviouslymentionedstimulationperiodsintofamiliarvoiceandunfamiliarvoice.ToexaminetheeectofrhymefamiliarityintheEG,werepeatedthedescribedprocedureanddividedstimulationperiodsintofamiliarandunfamiliarrhyme.Toexaminewhetherauditorystimulationaectstheautonomicnervoussystemreaction(measuredbyheartrate),thesleepstagingdataandthecorrespondingmeanheartrateswerematchedforevery30-secondepochanddividedintobaselineandstimulationperiods.Afterward,meanheartratesforAS,QS,andWduringbaselinesandstimulationperiodswerecalculatedforeverysubject.3.Results3.1.EectofAuditoryStimulationonSleep-Wake-StatesRepeatedmeasuresANOVAwiththeindependentvariablesandwithin-factorsbehavioralSTATE(QS,AS,W),PHASE(baseline,stimulation)andAGE(2weeks,5weeks)andthebetween-subjectfactorGROUP(prenatalstimulation/experimentalgroup=EG,noprenatalstimulation/controlgroup=CG)werecomputed.Thedependentvariablewasthetime(inseconds)spentwithinacertainbehavioralstate(W,AS,orQS).Importantly,theamountofoverallstagedsleepepochsistherebynotdierent BrainSci.2020,10,5116of11betweenbaselineandsimulationperiodsatbothrecordingstwo(t(33)=1.56,p=0.129)andfive(t(33)=0.749,p=0.459)weeksafterbirth.Furtheranalysesrevealedthat,overall,thetimespentinthethreebehavioralstateswasdierentforW,QS,andAS(F(1.53,49.03)=14.71,p<0.001,p.eta2=0.32),withmostofthetimespentinactivesleep,followedbywakeandquietsleep(fordetailspleaserefertosuppl.materials).Inaddition,thetimespentinthethreebehavioralstatestendedtobedierentbetweentheEGcomparedtotheCG,asindicatedbytheinteractionSTATE*GROUP(F(2,64)=2.82,p=0.067,p.eta2=0.08).Exploratorypost-hocindependentt-testrevealed(t(32)=1.89,p=0.068)thatthetimespentinAStendedtobehigherintheEGgroup(M=421.60,SE=32.10)ascomparedtotheCG(M=302.05,SE=62.48)andalsothetimespentinWtendedoveralltobelowerintheEG(t(32)=1.73,p=0.093;EG:M=143.15,SE=32.24vs.CG:M=256.36,SE=67.08).TheinteractionSTATE*PHASEindicatesanothertrendfortheEG(F(2,44)=2.89,p=0.066),andnosucheectfortheCG(F(2,20)=1.17,p=0.331,p.eta2=0.10)whenfocusingonthechangefrombaselinetostimulationforthethreebehavioralstatesofinterest(cf.Figure2).Exploratorypost-hocpairedt-teststherebyrevealedthatintheEGgroupthetimespentinWdecreased(t(22)=2.32,p=0.029)frombaseline(M=157.82,SE=33.53)tostimulationperiods(M=154.22,SE=32.16),whereasQSincreased(t(22)=2.32,p=0.030)frombaseline(M=77.61s,SE=21.12s)tostimulationperiods(M=116.73,SE=25.70).Non-parametricbootstrappinganalysis(1000repetitions)confirmsthatWisreducedintheEGfrombaselinetostimulation(t(22)=2.32,p=0.033),whereasQSisincreasedfrombaselinetostimulationt(22)=2.32,p=0.044)whichweinterpretascalmingeectofthestimulationinthe(auditorystimulation)familiarizedEG.Figure2.Behavioralstatesinresponsetoauditorystimulationininfantsprenatallyexposedto(experimentalgroup,EG)andunexposedto(controlgroup,CG)auditorystimulation.Figuresshowtheproportionsoftimespentinquietsleep(QS),activesleep(AS),andwake(W)statesduring(43min)baseline(silence)andduring(43min)auditorystimulationconditionsininfantswhowereprenatallyexposedtoauditorystimulation(A;EG;n=23)andwhowerenotexposed(B;CG;n=11).Notethatinfantsfamiliartoprenatalstimulation(EG;A)fellasleepmorelikely(lessW)aswellassleptdeeper(moreQS)duringauditorystimulationandgenerallyexhibitedlessWstatesatbaselineandstimulation.Ontheotherhand,unexposedinfants(CG,B)exhibitapatternindicatingnochangesinbehavioralstatesfrombaselinetostimulation.Curlybracketshighlightthesignificantpost-hoct-testcomparisons.*=statisticalsignificancep<0.05;Errorbarsrefersto1StandardErroroftheMean(SEM).3.2.EectofVoiceFamiliarityonSleep-Wake-StatesRepeatedmeasuresANOVAwiththewithin-subjectfactorsbehavioralSTATE(QS,AS,W),PHASE(baseline,stimulation),VOICE(familiar,unfamiliar),andAGE(2weeks,5weeks)revealedamaineectforSTATE(F(1.48,48.91)=20.47,p<0.001,p.eta2=0.38),butnointeractionforSTATE*VOICE(F(1.58,51.30)=1.71,p=0.196,p.eta2=0.05)orSTATE*PHASE*VOICE(F(1.35,44.58)=1.60,p=0.215,p.eta2=0.05;cf.SupplementalFigureS1),indicatingthatvoicefamiliaritydidnotinfluencetheamountoftimespentinthethreedierentbehavioralstates. BrainSci.2020,10,5117of113.3.EectofRhymeFamiliarityonSleep–Wake-StatesInadditiontotheeectofvoicefamiliarity,wealsotestedwhetherrhymefamiliarityduetoprenatalstimulationwasevidentattwoandfiveweeksafterbirth.Rhymefamiliaritywastestedusingarepeated-measuresANOVAwiththewithin-subjectfactorsbehavioralSTATE(QS,AS,W),PHASE(baseline,stimulation),RHYME(familiar,unfamiliar),andAGE(2weeks,5weeks)duringstimulationintheexperimentalgroup(EG).Analysisrevealedthatthetime(inseconds)overallspentinthethreebehavioralSTATEswasdierent(F(1.39,30.59)=35.23,p<0.001,p.eta2=0.62)withmoretimespentinAS.However,theinteractionbehavioralSTATE*RHYMEfamiliarity(F(1.44,31.70)=0.21,p=0.752,p.eta2=0.01;cf.SupplementalFigureS2)wasnotsignificant,indicatingthattimespentinbehavioralstatesdidnotchangeinresponsetofamiliarvs.unfamiliarrhymeexposureafterbirth.Likewise,thetimespentinthethreebehavioralSTATESdidnotchangeasafunctionofRHYMEfamiliarityfrombaselinestostimulation(PHASE*RHYME*STATE:F(2,44)=0.24,p=0.587,p.eta2=0.02).3.4.EectofAuditoryStimulationonPhysiologyInadditiontotheanalysisofbehavioralstateshiftsinresponsetothere-exposurewiththefamiliarandunfamiliarauditorystimuli,wecomplementedthepicturewithanalysesofbabies’physiology,namelyheartrate(HR)irrespectiveofrhymeandvoicefamiliarity.AnalysisofHRusingathree-wayANOVAwithTIME(93minperiods),AGE(2weeks,5weeks),andGROUP(EG,CG)revealedamaineectforGROUP(F(1,29)=8.99,p=0.006,p.eta2=0.24).Post-hocindependentt-testsindicatealowermeanHRintheEG(M=140.66,SD=7.81)ascomparedtotheCG(M=152.56,SD=10.71)duringbaselineandstimulationperiods.Additionally,comparingthefirsthalf(12min;BL1-ST2)andthesecondhalf(12min;BL3-ST4)oftherecordingsrevealedthattheEG,butnottheCG,reducedHR(t(20)=2.67,p=0.015,d=0.57)fromthefirsthalf(M=143.71,SD=8.99)tothesecondhalf(M=138.64,SD=8.83)oftherecording(cf.Figure3).Data,therefore,indicateshabituationoverthecourseoftherecordingexclusivelyfortheprenatallyfamiliarizedEGbutnotthe(auditorystimulation-)naïveandunexposedCG.Formoredetails,seeSupplementalTableS1.Figure3.Evolutionoftheheartrate(HR)overtime.ThetwotimelinesshowthemeanHRforthefourchronologicallyorderedbaseline(BL1,2,3,4)andstimulation(ST1,2,3,4)periodsof3mineachindependentofrhymeandvoicefamiliarity.Notethatinfantsfamiliar(EG)andunfamiliar(CG)withprenatalstimulationshowdistinctHR.EspeciallyinthesecondparttheEGseemstoshowhabituationanddecelerationoftheHRasrevealedbypaired-samplet-tests.Dataispooledfromthesamebabiesrecordedtwiceatweektwoandweekfiveofage.BLendreferstothelast3minbaselinefollowingfourbaselineandstimulationperiods.*=statisticalsignificancep<0.05;Errorbars=1SEM.4.DiscussionThepresentstudyaimedtoinvestigatewhethertheeectsofrepeatedauditorystimulationduringpregnancyarevisibleinthefirstweeksafterbirth.Specifically,wehypothesizedthatdailyprenatal BrainSci.2020,10,5118of11exposuretoanurseryrhymewouldleadto“recognition,”andthereforehabituation,tothatrhymetwoandfiveweeksafterbirth,asevidencedbymorecalmandrelaxednewbornsduringstimulationwiththefamiliarnurseryrhyme.Interestingly,wefoundnostableeectofstimulusfamiliarityonthenewborns’sleep-wakestates,neitherforrhyme,norvoicefamiliarityintheprenatallyexposedexperimentalgroup.However,whatwasfoundwasageneraleectindicativeof“fetalprogramming.”Weobservedthatonlythosebabieswhowerealreadyprenatallyexposedandfamiliarizedtoanauditorynurseryrhymeonadailybasis(experimentalgroup,EG)calmeddownwhentheserhymes(regardlessoffamiliarity)werereplayedafterbirth,whereastheprenatallyunstimulatednaïvecontrolgroupshowednosucheects.TheformerfindingintheEGwasalsoreplicatedinnewborns’physiologicalresponseswithaprogressiveslowingdownoftheheartrateacrosstherecordingperiodforthepre-exposedEGonly.Asimilarcalmingeectduringre-exposurewithauditorystimulationwasreportedearlierbyGranier-Deferreetal.[36],whofoundHRdecelerationsininfantsre-exposedtoafamiliar(prenatallypresented),butalsotoanunfamiliar,pianomelody.DailyprenatalexposuretoauditorynurseryrhymesseemstohavefamiliarizedbabiestosuchenvironmentsafterbirthastheunexposedCGshowedahigherpercentageofwakestatesduringexposurewiththenurseryrhymesaswellasagenerallyelevatedheartrate.ItistonotethatthisisnotduetoawakeningsduringauditorystimulationintheCG,butEGinfants(familiarizedprenatallywithauditorystimulation)generallyseemedcalmerandsleepierduringauditory(re-)exposure.ThisisinlinewithfindingsfromGonzalez-Gonzalezetal.[27],whoreportedthatnewbornsrepeatedlyexposedtoa(vibroacoustic)stimulusinuterohabituatedearliertothesamestimulusafterbirth.Muenssingeretal.[31]havealsoreportedhabituationtorepeatedlypresentedauditorystimulationeveninthird-termfetusesusingfetalMEG,suggestingthattheneuronalbasisforlearningisalreadypresentbeforebirth.Anotherlesslikelyexplanationfortheobservedcalmingeectcouldbe,thatnewbornsarelinkingthere-exposuretothenurseryrhymestotheprenatalauditorystimulationsetting,wheremothers-to-bewereinstructedtositcalmlyandrelaxwhilelisteningtotherhymesvialoudspeakers.Itispossiblethatthisimmobilerelaxedsettingwaslinkedtotheauditorystimulationandresultsinmorerelaxedfetusesand,therefore,morerelaxednewbornsatauditoryre-exposureafterbirth.Altogether,dailyauditorystimulationofourthird-trimesterfetusesmay,therefore,haveresultedinbasicmemoryfor,andquickerhabituationto,suchconditions.Infantsnotbeingfamiliarizedbeforebirth,ontheotherhand,mayreactwithirritationorattentionorientationtoenvironmental(speech-like)noise.Themostinterestingpointhereisthatprenatallyfamiliarizedinfantscalmeddownregardlessofstimulusfamiliarityand,additionally,despitetheauditorystimulusbeingsignificantlydistortedby,e.g.,theamnioticfluidduringthepre-natalstimulationperiodinutero.Thisfindingindicatesgeneralizationofprenatallylearnedmaterialinnewborninfants.However,whetherprenatallearningispossibledespiteorduetothespecialsignificanceofsleepformemoryformationduringtheperinatalperiodisstillaquestiontobeanswered.Somerecentstudieshaveshownthatmemoryinyounginfantsbenefitsfrom[46]andgeneralizesduringsleep[47]asearlyasthreemonthsafterbirth.Whetherperinatalsleepjustbeforeorafterbirthhassimilarsignificanceisyetunknownbutwouldbehighlyinterestingaspossiblystimulipresentedattwoweeksoflifeinourstudymayalreadybelearnedandrecognizedas“familiar”atourfive-weekfollow-up.Contrarytoearlierstudiesshowingstablecalmingeectsforthematernalvoice[13,14,35],wefoundnodierenceinthetimespentinwakeorsleepinthepresenceofthefamiliarmaternalvoicecomparedtoperiodswhentheunfamiliarfemalevoicewaspresent.Asmentionedintheintroduction,thematernalvoiceissometimesalsoreportedtobearousingfornewborns[11,12].Asinfantswereindierentbehavioralstatesatstimulusonset,andthereforemore(awake)orless(sleep)responsivetothestimulationwiththematernalvoice,weconcludethismightbeapotentialreasonforthemissingeectinourstudysample.Additionally,andalsocontrarytoanearlierstudy[33],wefoundnodistinctchangesinthetimespentinbehavioralstates(W,AS,orQS)totheprenatallypresentednurseryrhyme.Givenourcurrentanalysis,weconcludethatmemoryoftheprenatallyreplayednurseryrhymeisonlyencodedtoabasicdegree,whichisnotidentifiableatabehavioral(i.e.,sleep-wakestagetransition) BrainSci.2020,10,5119of11level.Furtheranalysisonbrain(EEG)andautonomicphysiology(ECG)level)isnecessarytoseeifsucheectsareevidentatamorefine-grainedphysiologicallevel.Asalimitationofourstudy,onecoulddiscusstherathershortdurationofourrecordings(~27min)andthereforealimitationofsegmentsavailableforeachandeverysleepstagepernewborn.However,inourexperience,thisproblemishardtocircumventinasampleofhealthynewbornsintheirhabitualhomeenvironment.First,itseemsimpossibletoplantherecordingsinsuchawaythattheinfantsareawakeandattentiveatthestartoftheexperimentalparadigmsasnewbornsasyoungastwotofiveweeksdonotyetexhibitregularsleep–wake-cyclesandconstantlyfallasleepirrespectiveoftimeofdayorenvironmentalstimulation.Inaddition,alreadythepreparationoftheEEG-system,theECGmontageandsettingupofthevideographyinthebabies’hometakestimeandappearstiringforthenewborns.Second,wealreadyusedcomfortablehdEEG-netswiththemainaimtosavevaluabletimeinEEGsetupandstillobservedthatnewbornsoftengotuneasyandwhinyafterabout30min(anearlierabandonedversionoftheparadigmlasted45min).Experimentallythe27minregistration,therefore,appearedasanoptimaltrade-obetweencalmandcomfortableinfantsandtheminimumamountofdataneededforsensibleECGandEEGanalysis.Asecondlimitationisthesamplesize,andinparticular,theunbalancedgroupsizesforexperimentalandcontrolgroups.Asdataisdiculttogatheratthisveryearlystageoflifeinhealthynewborns(here,wealreadyrecorded45babiestwiceintheirhomeenvironmentusingfullpolysomnographysetups),webelievethatthisisthebestthatbedonewithinacomprehensivepilotstudy.Finally,wecanonlyspeculatewhytheEG(familiarizedtothedailystimulation)startswithalowerheartrate(Figure3)alreadyinthebaseline.WebelievethattheEGmaysimplybelessarousedbytheenvironmentalnoiseduringthepreparationintheirhomeenvironmentastheymaybegenerallyless“environmentalnoisesensitive”aftertheirdailyexposuretotheloud(80dB)rhymestimulationbeforebirthoverthecourseofsixweeks.Inconclusion,ourpreliminarydataindicateshowprenatalexperienceor“fetalprogramming”mayhaveaneectonbehavioral(sleep/wakestates)andphysiological(heartrate)reactionsjustweeksafterbirthasevidentinlesswakingperiodsandHRhabituationtostimuliheardalreadyinthelasttrimesterbeforebirth.Parentsandsocietiesshouldbeawareofsucheectsandmayconsiderthisintheirparentingmethodsevenbeforebirth.Thisisnottosaythatweareinfavorofoverambitiousstimulationofthefetusinordertomaximizelearningevenbeforebirth.Ratherwecallintoattentionthatmuchofwhatafetusisexposedtobeforebirth—whetheritisparentalmovements,touch,music,orspeech—mayshapetheinfants’physiologyandlaterperceptionoftheworld.Inwhichexactwaywillbeworkforfurthergenerationsofdevelopmentalscientists.SupplementaryMaterials:Thefollowingareavailableonlineathttp://www.mdpi.com/2076-3425/10/8/511/s1,FigureS1.Eectofvoicefamiliarity;FigureS2.Eectofrhymefamiliarity;TableS1.Meanheartratesinexperimentalandcontrolgroup;Audiofiles:Kindchen_calm.wav,Butzemann_lively.wav.AuthorContributions:Conceptualization,M.S.andR.d.G.;methodology,M.S.andR.d.G..;validation,A.L.,R.d.G.andM.S.;formalanalysis,A.L.;investigation,A.L.andR.d.G.;resources,M.S.;datacuration,A.L.andM.S.;writing—originaldraftpreparation,A.L.;writing—reviewandediting,M.S.andA.L.;visualization,A.L.;supervision,M.S.;projectadministration,A.L.Allauthorshavereadandagreedtothepublishedversionofthemanuscript.Funding:AdelheidLangandRenatedelGiudiceweresupportedbytheDoctoralCollege“ImagingtheMind”(FWF;W1233-G17).Acknowledgments:WethankJulia-MariaPletzer,AndreasKovacs,SindySiebertandBarbaraFüttererfortheirgreateortandassistanceinrecordingnewborns’ambulatoryhdEEGandMalgorzataWislowskaforhersupportofallkinds.ConflictsofInterest:Theauthorsdeclarenoconflictofinterest.References1.Grigg-Damberger,M.Thevisualscoringofsleepininfants0to2monthsofage.J.Clin.SleepMed.2016,12,429–445.[CrossRef][PubMed]2.Kisilevsky,B.S.;Low,J.A.Humanfetalbehavior:100yearsofstudy.Dev.Rev.1998,18,1–29.[CrossRef] 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