Clinical study of PCSO-524 (ANTINOLäas neutraceutical · Clinical study of PCSO-524 (ANTINOLä) as...
Transcript of Clinical study of PCSO-524 (ANTINOLäas neutraceutical · Clinical study of PCSO-524 (ANTINOLä) as...
ClinicalstudyofPCSO-524(ANTINOLä)asneutraceuticalincanineallergicskindisease
Introduction
PCSO-524 is an extract from New Zealand green lipped mussel (Perna canaliculus). Itcontains several classes of lipid. PCSO-524 contains two of the long chain omega-3polyunsaturated fatty acid (PUFAs); eicosapentaenoic acid (EPA) and docosahexaenoic acid(DHA).PCSO-524iscomposeof13%EPA,21%DHAandabout30%cholesterol.1Inaddition,italso contains novel ω-3 PUFAs; 5,9,12,15-octodecatretraenoic acid, 5,9,12,16-nondecatertraenoic acid, 7,11,14,17-eicosatetraenoic acid, and 5,9,12,15,18-heneicsapententaenoicacid.2EPA,DHAandtheo-3PUFA7,11,14,17-eicosatetraenoicacidhavesimilar structure to arachidonic acid which is the precursor of the inflammatory agents,prostaglandinsandleukotrienes.2
TherehavebeenmanyreportsinhumanstudyaboutP.canaliculuseffects.EPAandDHAhavebeen reported to reduce the levelsof inflammatoryprostaglandins and leukotrienesascompetitive substrates for the cyclooxygenase enzyme (COX) and the lipoxygenase enzyme(LOX). IthasbeenshownthatPCSO-524can inhibitPGE2productionandabolish leukotrieneformation.3,4Anotherstudyshowedanti-histamineactivityofP.canaliculus.5
Inanimalstudy,ithasbeenshownthatPCSO-524hasmildanti-inflammatoryactivitytoreduceswellingininducedarthritis.4OtherstudyreportedthatPCSO-524orglucocorticoidalonedidnothaveclinicalbenefitonreducing inflammationbutcombinedtreatmentof these twosubstancescanreducepawswelling.6However,therehavebeennoreportedtocompareclinicaleffects of PCSO-524 and glucocorticoid in standard model or arthritis.2 The effects of P.canaliculushasalosobeendoneinthecollagentype-II-inducedseverepolyarthritis inratandmouse.TheresultsshowedthatP.canaliculuscanreduceincidenceofarthritisfrom58%fromcontrolgroupto17%inP.canaliculustreatedgroup.7ManystudiesofPCSO-524effectsindogswith arthritis have been reported. Bui and Bierer (2001) reported beneficial effects of P.Canaliculusthatcantreatmentgroupshowedanimprovementofosteoarthritiswhencomparedtountreateddogs.8ThisstudyshowedthatPCSO-524hasbeneficialeffectsonArthritisscores(mobility, pain, swelling and crepitus). Another report showed effectiveness of green lippedmuscleextractindogswithgenerativejointdisease.9Thisstudyshowedthatgreenlippedmusselextracthavebeneficialeffectstoimprovedmusculoskeletaldysfunctionby67%ofthe81lamedogs with no side effects throughout the treatment period. This study also showed clinicalresponse was seen at 56 days after treatment. A comparison between PCSO-524 and non-steroidalanti-inflammatorydrug(carprofen)wasdonein45dogswithosteoarthritis.10Dogsinplacebogroupshowed20-40%improvementinpain,mobilityindexandlocomotionafter8-weektreatment period while dogs treated with PCSO-524 and carprofen showed 67% and 86%improvementinpainVAS.SocarprofenwasshowntobemoreeffectivethanPCSO-524butitsadverse effects, especially with long term use, are of concern. Even though many studiesreported anti-inflammatory effects of PCSO-524 but no clinical study in inflammatory skindiseaseshavebeenreported.
Allergicdermatitisreferstoanyinflammatoryskindiseasescausedbyanytypesofallergy.Itiscausedbyvariousantigensinducingimmunologicalreactions.Theseverityandprogressionof clinical signs depend on etiologic factors. The clinical presentation of hypersensitivity caneitherbeshort-livedorlifelong.Insmallanimalpractice,severaltypesofallergicdiseasesaredocumentedbut themost common in dermatology practice includes atopic dermatitis, foodhypersensitivity,contacthypersentitivity,andfleaallergicdermatitis.11-13Thecharacteristicsofallergicdermatitisarepruritusandsubsequentinflammation.Therefore,allergicdermatitisisagood candidate to study anti-inflammatory effects of PCSO-524 for the treatment of skindiseases.Objectives
1.TostudythebeneficialeffectsofPCSO-524(ANTINOLä)asaneutraceuticaltoimproveskinandcoatcondition
2. To investigate therapeutic effects of PCSO-524 (ANTINOLä) in canine allergic skindiseases
Materialsandmethods
Animals The31dogsincludedinthisstudywereselectedfromdogswithskindiseasespresentingtotheSmallAnimalHospital,FacultyofVeterinaryScience,ChulalongkornUniversity.Alldogshadclinicalhistoriesconsistentwithchronicpruritis,includingsomedegreeoferythema,focalormultifocalalopecia,excoriation,andrecurrentbacterialoryeastinfection.Thedogsremainedundertheirowner’scarethroughoutthestudyperiod.
InclusioncriteriaThedogsunderwentageneralanddermatologicalexaminationconductedbythesame
veterinarian,toidentifyskinlesions.Hematologicalandbiochemicalanalysiswereperformedtoevaluatehealthstatus.Cytologyexaminationandbacterialcultureandsusceptibilitytestwerealso done to identify the presence of yeast and bacterial infection and select appropriatetreatmentforaffecteddogs.Allergicskindiseasewasdefinedusingacriterionofchronicpruritusandinflammationofskinwithotherskindiseasesexcludedbasedonhistory,clinicalsigns,skinscraping,cytologyexamination,hematology,andbloodchemistryprofiles.13
TreatmentprotocolDogsincludedinthisstudywereclassifiedinto3treatmentgroupsbasedonseverityof
dermatologicalsigns;mild,moderateandsevere.Theclassificationwasmadeonthebasisofdegreeofpruritus,distributionandextensionofthelesionsandskinandcoatcondition.14forethicalreasons,therewasnocontrolgroupwithouttreatment.Group1Milddegree: PCSO-524Group2Moderatedegree: PCSO-524+antihistamine(hydroxyzinehydrochloride)Group3Severedegree: PCSO-524+prednisolone
All dogs were treated with appropriate symptomatic therapy to control secondaryinfection in order to reduce other involving factors apart from allergic reaction. This wasprescribedbeforestartoftheexperimentorduringexperimentalperiodifsecondaryinfectionwas diagnosed up on clinical examination and cytology examination. Adverse effects weremonitoredclinicallybyveterinariansandnotedifanyadverseeffectswereseenbyownersorveterinarians.
Doseprescription
• PCSO-524: All the dogs received PCSO-524 of loading dose for 2 weeks followed bymaintenancethroughouttheexperiment.Loadingdosewasprescribedas1capsule/10kgBWtwiceadayandmaintenancedosewas1capsule/10kgBWonceaday.
• Antihistamine(hydroxyzinehydrochloride)2-4mg/kgbid
• Prednisolone:0.5-1mg/kg/d for7days then taperdown to0.25mg/kg/donceadayeveryotherday
• Cephalexin:25-30mg/kg/dbid
MonitoringofthedogsAlldogswerescheduledtorevisitandbeevaluatedat2weeks,4weeks,8weeks,12
weeksand16weeksaftertreatment.Evaluationofclinicalresponsei.e.improvementofclinicalsigns, response to therapy and skin and coat condition were assessed by owners andveterinarians.
VeterinaryevaluationGrossobservationanddermatologicalexaminationweredonebyveterinarianstoassess
effects of the treatment on each visit. Two veterinarian subjectively assessed 4 parameters;erythema, lichenification,excoriationandself-inducedalopeciausingcanineatopicdermatitisextentandseverityindex03(CADESI-03)atweek0,2,4,8,12and16.Thescoresassignedbyveterinarianweresummedtoformaveterinary-assessedindexscore.
OwnerevaluationOwners answered a questionnaire on general history of the dogs including gross
appearanceofthedogs,generalskinandcoatcondition,degreeofpruritus,ageofonsetandprevioustreatmentwererecordedonfirstvisit.Oneachfollowingvisit,observationofownersongeneralskinandcoatconditionanddegreeofpruritus(pruritusvisualanalogscale,PVAS)werenotedtoformanowner-assedindexusedadescriptivescaleof0-10.14
MajoroutcomecriteriaandstatisticalanalysisThemajoroutcomecriterionwastheproportionofdogswithimprovement,stableand
worseningofclinicalsignsperformedateachvisit.Statisticalanalysiswerecarriedoutbyusingthe Statistical Analysis System (SAS) software version 9.0 (SAS Inst. Inc.,Cary, NC, USA).DescriptivestatisticswereobtainedusingMEANSandFREQprocedures.AnalysisofvariancewasappliedusingtheMIXEDandGLMprocedures.Traitsanalyzedwerescoreof improvement inskinandcoatcondition,degreeofpruritusandlesionscores.One-wayANOVAswereusedtoevaluatechangesinCADESI-03andpruritusscoresbetweenthetreatmentgroupsfromweek0to16.Theproportionsofdogsineachtreatmentgroupsthatshowedimprovementatweek0,2, 4, 8, 12 and 16 were assessed. Chi-square test was used to analyze differences in theproportionsofdogsthatshowimprovement.ValueswithP<0.05wereregardedasstatisticallysignificant.
Week02481216
Evaluationbyowner
• Generalcoatandskincondition
• Degreeofpruritus
Evaluationbyveterinarians
• Grossobservation/clinicalsigns
• Hematologyandbloodchemistryprofiles
• Cytologyexamination
• Lesionscore(CADESI-03)
Treatmentprotocol
Loadingdose Maintenancedose
Result
Breeddistribution Therewere31dogsdiagnosedashavingallergicskindiseases.SeveralbreedsincludedinthestudywiththehighestnumberinShihtzu(n=9).Theotherbreedsalsoincludedinthisstudywere Poodle (n=6), Pomeranian (n=6),Mixed breed (n=3),Miniature pincher (n=2),Maltese(n=1),Englishcocker(n=1),Pug(n=1),Bangkaew(n=1)andChihuahua(n=1). Sex Fromtotalof31dogs,thenumbersofmaleandfemaledogwere15(48%)and16(52%).Thenumberofdogsandpercentagewerenotstatisticallydifferent. Comparisonbetween3treatmentgroups
Numberofdogswithimprovementinskinandcoatconditioningroup1,2and3are9of12,3of9and5of10 respectively.Group1showedhigherpercentage (75%)comparedwithgroup2(33.33%)andgroup3(50%).Similarly,numberofdogswithreductioninpruriticscore(PVAS)wasalsohigherinimprovementingroup1(50%)withlesseffectsingroup2(44.44%)and3(40%).Themoreprominenteffectsoflesionscore(CADESI-03)improvementwereobservedwithsignificanthigherpercentageascomparedwiththeothertwogroups.Group1showedtheshortesttimeforimprovementseenwhencomparedwithgroup2and3.Timeat improvementseen inGroup1,2and3were5.33+1.56,7.33+2.70and12+2.09,respectively. Adverseevents One dog in group 3 had diarrhea 2 weeks after treatment. Diarrhea was mild andtransient.Therewasnoevidenceofserioussideeffectsormoderateorseveregastrointestinalevents.Inparticular,nochangesinhematologyandbloodchemistryprofilesindogstreatedwithPCSO-524alone.Somedogs ingroup3 (PCSO-524+prednisolone)had increased liverenzymelevels.
Table1Subjectcharacteristics
Subject
Age(year)
Sex
Breed
Ageofonset
(year)Group1 1
23456789101112
74123889
10.559510
FMMMMFsMMFMMF
PoodleShihtzuShihtzuChihuahuaShihtzuShihtzuPomeranianPomeranianPomeranianMaltesePomeranianShihtzu
4-71-31-31-34-74-74-74-74-74-71-34-7
Group2 123456789
54887.5144919
FFFsMFMFMM
ShihtzuPugMixedPomeranianPooleBangkaewMiniaturepincherShihtzuMiniaturepincher
1-31-3<14-74-71-3<14-7>7
Group3 12345678910
810881312613149
MFMFFFFMFF
MixedPoodlePomeranianMixedPoodlePoodleShihtzuPoodleEnglishcockerShihtzu
4-71-3>74-74-74-74-7>7>74-7
M=male;F=female;Fs=spayedfemale
Figure 1 Diagram shows breed distribution of 31 dogs included in the study; Shih tzu (n=9),Poodle (n=6), Pomeranian (n=6),Mixedbreed (n=3),Miniaturepincher (n=2),Maltese (n=1),Englishcocker(n=1),Pug(n=1),Bangkaew(n=1)andChihuahua(n=1).
29%
20% 19%
10%
7%
3% 3%
3% 3% 3%
Breed
Shihtzu Poodle Pomeranian MixedbreedMiniaturepincher Maltese Englishcocker PugBangkaew Chihuahua
Figure2Diagramshowssexof31dogsdiagnosedashavingallergicskindiseaseinthisstudy;male(n=15)andfemale(n=16).
48%
52%
Sex
Male
Female
Figure3Ageofonsetnotedinallsubjects(31dogs)wereclassifiedinto4ageranges;<1years(n=2),1-3years(n=8),4-7years(n=17)and>7years(n=4).
6%
26%
55%
13%
Ageofonset
<1year
1-3years
4-7years
>7years
Figure4Clinicalresponse(skin&coatconditionanddegreeofpruritus)inallergicdogstreatedwithPCSO-524alone(n=12).
0
1
2
3
4
5
6
7
8
9
Improved Stable Worseskin&coatcondition 9 3 0
Degreeofpruritus 6 5 1
Num
bero
fdog
sGroup1:PCSO-524
Figure5Clinicalresponse(skin&coatconditionanddegreeofpruritus)inallergicdogstreatedwithcombinationtherapyofPCSO-524andantihistamine(n=9).
0
1
2
3
4
5
6
Improved Stable Worseskin&coatcondition 3 6 0
Degreeofpruritus 3 6 0
Num
bero
fdog
sGroup2:PCSO-524 +Antihistamine
Figure6Clinicalresponse(skin&coatconditionanddegreeofpruritus)inallergicdogstreatedwithcombinationtherapyofPCSO-524andprednisolone(n=10).
0
0.5
1
1.5
2
2.5
3
3.5
4
4.5
5
Improved Stable Worseskin&coatcondition 5 4 1
Degreeofpruritus 4 5 1
Num
bero
fdog
sGroup3:PCSO-524 +Prednisolone
Figure7Canineatopicdermatitisextensionandseverityindex(CADESI-03)scoreofallergicdogstreatedwithPCSO-524alone(n=12),combinationtherapyofPCSO-524andantihistamine(n=9)andPCSO-524andprednisolone(n=10).
0
2
4
6
8
10
12
Improved Stable Worse
PCSO-524 11 1 0
PCSO-524+AH 3 5 1
PCSO-524+Pred 4 4 2
Num
bero
fdog
s
CADESI-03score
Table2Percentageofimproveddogsandtimeatimprovementseenforevaluatedvariables,per
group
Numberofdogswithimprovement(%)
Timeatimprovementseen
(weeks)
Group1PCSO-524(n=12)
Group2
PCSO-524+AH(n=9)
Group3
PCSO-524+Pred(n=10)
Group1PCSO-524(n=12)
Group2
PCSO-524+AH(n=9)
Group3
PCSO-524+Pred(n=10)
Skin&coatcondition
9/12(75%)
3/9
(33.33%)
5/10b,c(50%)
5.33+1.56c
7.33+2.70
12+2.09a
Degreeofpruritus(PVAS)
6/12(50%)
4/9
(44.44%)
4/10(40%)
4+0.95
4.67+1.40
5+1.17
Lesionscore(CADESI-03)
11/12b,c
(91.67%)
3/9a
(33.33%)
4/10a
(40%)
4.55+1.28c
4.67+2.45
10.50+2.12a
asignificantdifferencebetweenwhencomparewithgroup1bsignificantdifferencebetweenwhencomparewithgroup2csignificantdifferencebetweenwhencomparewithgroup3
Table3Acomparisonof3parameter(mean+SEM);skinandcoatcondition,degreeofpruritus
andlesionscorein3treatmentgroupsatweek0,2,4,8,12and16
Week
Mean+SEM
0 2 4 8 12 16
Skin&coa
tcond
ition
Group1PCSO-524(n=12)Group2PCSO-524+AH(n=9)Group3PCSO-524+Pred(n=10)
6+0.67
7+0.77
6+0.73
5.7+0.61
6.13+0.68
6.00+0.68
3.91+0.64
5.22+0.74
5.1+0.71
2.67+0.56c
4.29+0.73
5.30+0.61a
2.89+0.66b
5.40+0.88a
4.50+0.69
1.88+0.63
5.50+0.72
4+0.67
Degreeof
pruritu
s(PVA
S)
Group1PCSO-524(n=12)Group2PCSO-524+AH(n=9)Group3PCSO-524+Pred(n=10)
5+0.72
6.89+0.83
6.1+0.79
4.4+0.59
6.38+0.67
5.75+0.67
3.5+0.63
5.33+0.73
5.1+0.69
2.33+0.6b,c
5.00+0.85a
4.9+0.66a
2+0.58b,c
5.6+0.73a
5.13+0.58a
2+0.78b
4.67+0.90a
3.85+0.84
Lesion
scores
(CAD
ESI-0
3)
Group1PCSO-524(n=12)Group2PCSO-524+AH(n=9)Group3PCSO-524+Pred(n=10)
250+52.51
254+60.63
359+57.51
196+53.17
326.67+68.64
255+59.44
89.75+36.77b,c
211.56+42.46a
211.30+40.28a
94.22+26.75b,c
124.57+30.34a
231.30+25.38a
75.89+32.18b,c
206.14+36.49a
232.50+34.13a
37.25+34.31b,c
177.57+36.67a
240.71+36.67a
asignificantdifferencebetweenwhencomparewithgroup1bsignificantdifferencebetweenwhencomparewithgroup2csignificantdifferencebetweenwhencomparewithgroup3
Discussion
Allergicdermatitisisdefinedasanyinflammatoryskindiseasescausedbyanytypesofallergy.Itcanbecausedbyavarietyofallergeninducingimmunologicalreactionofanimals.Inthisstudy,allergicdogsreferstodogswithclinicalsignsofatopicdermatitisandfoodallergyasother diseaseswere ruled out by history, physical examination, cytological examination andbloodtests.Thediagnosisfollowed2010clinicalpracticeguidelinesfromtheinternationaltaskforceoncanineatopicdermatitis.14
Eicosapentaenoicacid(EPAanddocosahexaenoicacid(DHA)arethemainactiveportionsof fish oil. their anti-inflammatory effects are possibly due to inhibition of eicosanoids andcytokinesproduction.15AnextractfromNewZealandgreenlippedmussel(Pernacanaliculus),PCSO-524 also contains polyunsaturated fatty acid (PUFAs); eicosapentaenoic acid (EPA) anddocosahexaenoicacid(DHA)includingnovelω-3PUFAs(7,11,14,17-eicosatetraenoicacid).1,2Manystudiesshowedanti-inflammatoryactivityofthesePUFAsandalsoanti-histamineactivity.4Fishoilhasbeenshown tohavehigher inhibitoryeffectsonCOX-1andCOX-2productionascompared to PCSO-524. However, hydrolysed form of PCSO-524 showed similar inhibitoryactivity on COX enzyzmes production.16 PCSO-524 has beneficial effects on reducinginflammation in animal study. It hasbeen shown to reduce swelling in arthritis.3 In addition,PCSO-524hasalsobeenshowntohaveanti-inflammatoryeffectswhenusedincombinationwithglucocorticoid.6 The anti-inflammatory effects was absent when animals were treated withPCSO-524orglucocorticoidalone.5AnotherstudyshowedtheeffectivenessofcombinationofPCSO-524,EPAandDHAinthetreatmentofrheumatoidarthritis.17However,thestudyofPCSO-524effectsonskindiseaseshasneverbeeninvestigated.
From total of 31 dogs, the proportion ofmale and female dogs diagnosed of havingallergicdermatitiswasnotsignificant.Therewere15maledogs(48%)and16femaledogs(52%).Theresultscorrelatedwithpreviousstudythatallergicdermatitishasnogenderpredisposition.14
Dogsingroup1,treatedwithPCSO-524only,showedsignificantimprovementofskinandcoatconditioncomparedwithgroup2and3.Thiswasevaluatedbytheowners,withscorerangefrom0to10basedonskinandcoatgrossappearance.Similarly,numberofdogswithreductioninpruriticscore(PVAS)wasalsohigheringroup1(50%)withlesseffectsingroup2(44.44%)and3 (40%).Theresults indicatedthatdogswithmilddegreeofallergicdermatitisseemtohavebenefitfromPCSO-524byimprovingskinandcoatconditionandreducingdegreeofpruritus.Dogswithmoderatedegree(group2)andseveredegreeofallergicdermatitis(group3)didnothavebeneficialeffectsbyownerevaluationresults.Theimprovementobservedingroup1waspossibly due to anti-inflammatory and anti-histamine activity of PCSO-524.2-4 Failure oftreatmentsingroup2and3couldbeacombinationofdegreeofseverity,presenceofrecurrentbacterialandyeastinfectionandchronicityofthedisease.Olivryetal.(2010)14suggestedthatclinical signsofallergicdogs,e.g.urticariaorpruritus,mightbepresentwithorwithout skinlesion.ItwasreasonableforDogsingroup1withmilddegreeofallergytohaveimprovedclinicalappearance.Dogsingroup2and3withmoderateandseveredegreeofallergymighthavehigherdegreeof inflammationorhypersenstitivity, inwhich, combination therapywas incapableoftreatingthedogs.Allallergicdogsinthisstudymightrepresentatopicdermatitisorfoodallergy
orboth.Therelationshipbetweenatopicdermatitisandfoodallergyhasbeencontroversialbutitissuggestedfoodcomponentsmightcausetheflareofatopicdermatitis.Thisincludesurticariaorprurituswithorwithoutskinlesion(9ofguideline).Therefore,dietaryfactorscouldbeonefactoroftreatmentfailure.Moreover,dogsingroup2and3withhigherdegreeofseverityhadmorefrequentrecurrenceofbacterialoryeastinfection.Thisisamajorflarefactorsinallergicdogs.14
EvaluationbyveterinarianalsoshowedconsistentresultsofPCSO-524benefitonreductionin lesionscore (CADESI-03).The improvementof lesionsscoreswasobservedwithsignificanthigherpercentageingroup1comparedwiththeothertwogroups.Thetimeat improvementseenwasalsoobservedto investigatepropertreatmentperiodforeffectiveresults.Averagetimetohaveimprovementingroup1,2and3were5.33+1.56,7.33+2.70and12+2.09weeks,respectively.Theresultswereconsistentwithpreviousstudy.18Conclusion PCSO-524mighthavebeneficialeffectsforthetreatmentofcanineallergicskindisease.Theimprovementinskinandcoatcondition,degreeofpruritusandlesionscoreswasobservedindogswithmilddegreeofallergicskindiseasebutnotindogswithmoderateorsevereallergicdermatitis.Othercontributingfactors,i.e.secondarybacterialoryeastinfection,skinandcoathygieneandcausingallergensshouldbeofconcernandwell-controlledinmoreseverecases.References
1. Murphy KJ, Mooney BD, Mann NJ, Nichols PD, Sinclair AJ. Lipid, FA, and sterolcompositionofNewZealandgreen lippedmussel (Pernacanaliculus)andTasmanianbluemussel(Mytilusedulis).Lipids2002;37:587–95.
2. DoggrellSA.Lyprinol-IsItaUsefulAnti-InflammatoryAgent.EviCompandAltMed2011;1-7.
3. TreschowAP,HodgesLD,WrightPFA,WynnePM,KalafatisN,MacridesTA.Novelanti-inflammatoryo-3PUFAsfromtheNewZealandgreen-lippedmussel,Pernacanaliculus.CompBiochemPhysiol2007;147:645–56.
4. Whitehouse MW, Macrides TA, Kalafatis N, Betts WH, Haynes DR, Broadbent J. Anti-inflammatory activity of a lipid fraction from the NZ green-lipped muscle.Inflammopharmacology1997;5:237–46.
5. KosugeT,TsujiK,IshidaH,YamaguchiT.Isolationofanantihistaminicsubstancefromgreen-lippedmussel(Pernacanaliculus).ChemPharmBull1986;34:4825–8.
6. WhitehouseMW, Butters DE. Combination anti-inflammatory therapy: synergism in rats ofNSAIDs/corticosteroids with some herbal/animal products. Inflammopharmacology 2003;11:453–64.
7. LawsonBR,BelkowskiSM,WhitesidesJF,DavisP,LawsonJW.Immunomodulationofmurinecollagen-inducedarthritisbyN,N-dimethylglycineandapreparationofPernacanaliculus.BMCComplementAlternMed2007;7:20.
8. Bui LM,BiererTL. Influenceofgreen-lippedmussel (Pernacanaliculus) inalleviating signsofarthritisisdogs.VetTher2001;2:101–11.
9. Hielm-BjÖrkman,TulamoR-M,SalonenH,RackallioM.EvaluatingcomplementarytherapiesforcanineosteoarthritispartI:green-lippedmussel(Pernalcanaliculus).eCAM2007doi:10.1093/ecam/nem136.
10. Pollard B, Guilford WG, Ankenbauer-Perkins KL, Hedderley D. Clinical efficacy andtolerance of an extract of green-lipped mussel (Perna canaliculus) in dogs presumptivelydiagnoseswithdegenerativejointdisease.NewZealandVeterinaryJournal2006;54:114–8.
11. ScottDW,MillerJH,GriffinCE.Hypersensitivitydisorders. In:MullerGH,KirkRW,eds.Smallanimaldermatology.6thed.Philadelphia,Pa:WBSaundersCo,2001:571-650.
12. Willemse T. Atopic skin disease: a review and reconsideration of diagnostic criteria. JSmallAnimPract1986;27:771-778.
13. OlivryT,MarsellaR,IwasakiT,MuellerR(2007):ValidationofCADESI-03,aseverityscaleforclinicaltrialsenrollingdogswithatopicdermatitis.Vet.Dermatol.18,78-86.
14 Olivry T, DeBoer DJ, Favrot C, Jackson HA,Mueller RS, Nuttall T, Prélaud P (2010)Treatment of canine atopic dermatitis: 2010 clinical practice guidelines from theInternationalTaskForceonCanineAtopicDermatitis.VetDerm2010;3:233-248.
15.CalderPC.n-3polyunsaturatedfattyacids,inflammationandinflammatorydisease.Am
JClinNutr2006;83(6Suppl):1505S–19S16.McPheeS,HodgesLD,WrightPFA,WynnePM,KalafatisN,HarneyDW,MacridesTA.
Anti-cyclooxygenaseeffectsoflipidextractsfromtheNewZealandgreen-lippedmussel,Pernacanaliculus.CompBiochemPhysiolPartBBiochemMolecBiol2007;146:346–56.
17.LauCS,ChiuPKY,ChuEMY,ChenyIYM,TangWM,ManRYK,HalpernGM:Treatmentof knee osteoporosis with Lyprinol_, lipid extract of the green-lipped mussel – adoubleblindplacebo-controlledstudy.ProgrNutr2004;6:17–31.
18.VaughnDM,ReinhartGA,SwaimSF,LautenSD,BoudreauxMK,SpanoJS,HoffmanCEandConnerB.EvaluationofEffectsofDietaryn-6ton-3FattyAcidRatiosonLeukotrieneBSynthesisinDogSkinandNeutrophils.VetDerm1994;5(4):163-173.