w w w. s b f g n o s i a . o r g . b r / r e v i s t a
Aspidosperma
subincanum
II.
Usefulness
of
uleine
and
ribonucleic
fragments
in
the
treatment
of
AIDS
patients
Dominique
Maes,
Roland
Maes
∗ Parabolicbiologicals,Beauvechain,Belgiuma
r
t
i
c
l
e
i
n
f
o
Articlehistory: Received24May2014 Accepted4November2014 Availableonline11February2015
Keywords: Uleine HIV Tri-therapy
RibonucleicAcidOligomers(RNO)
a
b
s
t
r
a
c
t
Aidspatientsweretreatedduringayearwiththreedifferentfoodsupplementscommercially avail-able:para-pau-aspido(AspidospermasubincanumMart.exA.DC.,Apocynaceae);2Leid(nucleicacidsand cytokines);andParaImmuno(propolis,pollenandroyaljelly).Allfoods,giveneitheraloneorin com-bination,provedusefultoallAIDSpatientswhoreceivedthesupplements,betheseundertri-therapy (Triomine:stavudine,lamivudine,névirapine)orleftunattended.
©2014SociedadeBrasileiradeFarmacognosia.PublishedbyElsevierEditoraLtda.Allrightsreserved.
Introduction
The treatment of AIDS patients rests on the administration
ofthreechemicalsdesignedtoreducetheviralload.TheHIV is
immunodepressivebut noeffortismadeduringchemical
treat-menttostimulatethemultiplicationandactivityoflymphocytes. Thisstimulationwouldhoweverbenefitthepatientsbecausesome
drugs are known to be immunosuppressive, which add to the
immunodepressioninducedbythepathogen.Meansexistto
stimu-latetheimmuneresponse,eitherbyinducingthesynthesisofnitric
oxidebylymphocytesandothercells,orelse bypromotingthe
multiplicationoflymphocytes.
Uleine(1),presentinHimatanthuslancifoliusandAspidosperma subincanumMart.exA.DC.,Apocynaceae,havebeenextensively studied(Seidletal.,2010).Itscapacitytopromotethesynthesisof nitricoxide(Souzaetal.,2007)makesoftheseplantscandidates foruseasanadjuvantinthetreatmentofpatientssufferingfrom
animmune-depression.ThebarkofAspidospermasubincanumhas
beenpurifiedandthefoodsupplementrichinuleinesoobtained hasbeendemonstratedinnocuous(Federlinetal.,2014).
NucleicacidoligomershavebeenshowntoinhibitinvivoShope fibromaandvacciniaviruses(Beljanskietal.,1975).Thiseffectwas tracedtoastimulationofthemultiplicationoflymphocytesand platelets(BeljanskiandPlawecki,1979).Lymphocytepopulations
depletedinvivobycyclophosphamiderespondedtoastimulusof
ribo-oligo-nucleotides(Beljanskietal.,1983).Thispositiveeffectof
∗ Correspondingauthor.
E-mail:anda.mars@wanadoo.fr(R.Maes).
regenerationoflymphocytesbynucleicacidsobservedinvivowas verifiedontwomalignantlymphomacases(Donadioetal.,1991).
These food supplements would appear useful to help the
immune system of immuno depressed patientsas tuberculosis
and Aids patientstocounter the immunodepressive activityof
thepathogensaswellastheimmunodepressiveactivityofsome
drugsusedtoeliminatethepathogen.Thishypothesiswasverified byadministeringthesupplementsduringayeartoAIDSpatients. Inaddition,athirdfoodsupplementconsistingofpropolis,royal jellyandpollen,knowntostimulatetheimmunesystem,wasalso offeredtotheclinician,enablinghertomakeacompletestudyof
availableimmunostimulators.
Theclinicianfocusedontheimprovementofthehealthofher
patientsbyverifyingtheactivityofa combinationof food
sup-plements given topatientseither under chemical treatmentor
else,duetolackofresources,leftunattended.Lackofresources restrictedthemonitoringinthecourseoftimeoftheeffectsofthe
treatments,toweightandlymphocytenumbers.
Thedetailedrawresults(66pages)areavailable(Bouillonand
Ndayikunda,2006).Theauthorsanalysedandexploitedthem.
MaterialsandMethods
Uleinesourceandtheimmunostimulant
Uleine(1)wasextracted fromtheplantAspidosperma
subin-canumMart.exA.DC., Apocynaceae,and presentedasthefood
supplement“Para-pau-aspido”bytheCompanyParabolic
Biologi-cals(Beauvechain,Belgium)undertwodifferentconcentrations:
PP20 (20mg total alkaloids)and PP 80 (80mg total alkaloids).
http://dx.doi.org/10.1016/j.bjp.2014.11.011
Asupplyoftheproductssufficient tocovera yearoftreatment
wasdonated.Thefoodsupplement“ParaImmuno”elaboratedby
theCompanyParabolicbiologicalswasmadeofpropolisextracted frombee-hives,RoyalJellyandpollen.Thisproductwasdonated inamountssufficienttocoverayearoftreatment.
2Leidispreparedbythelaboratory“LaboratoireLabo-Life”.This
immunostimulanttobetakensub-linguallyismadeofcytokines,
ribonucleic acids, deoxyribonucleic acids and TGFbeta, and the
productwasdonatedinamountssufficienttocoverayearof treat-ment.
Triomine530:stavudine30mg,lamivudine150mg,névirapine
20mg,forpatientswhoseweightswasinferiorto60kg.
Triomine540:stavudine40mg,lamivudine150mg,névirapine
200mg,forweightsover60kg.
Clinicalstudies
Numberofsubjectstreated waseighty,ofwhomsixtywere
includedinthisstudy.Theywereorphansattheorphanage“Maison Shalom”,locatedintheBurundi.
The files of the patients were a year old when the study
wasinitiatedin2006andmanyshowedanunfavourableclinical
symptomatologyandopportunistinfections,i.e.chronicdiarrhoea,
persistent fever, generalised lymphadenopathies, diverse
pneu-mopathies,mycoses,generaliseddermatoses,zona,alterationof
thegeneralstate,lossofweightandasthenia.Nearlyallpatientshad
conservedtheirsymptomatologywhenthetreatmentwith
supple-mentsstartedandsomehadworsenedanddevelopedadditional
opportunistinfections.
Amongthesubjectsundertri-therapysincesixmonths,some
showed no clinical change, others had developed new
oppor-tunistillnessesandotherswereintoleranttothetri-therapy.Those
patientsundertri-therapywhoshowednoimprovementandwho
had developed opportunistic infections that werenot observed
beforethechemicaltreatment,wereincludedinthestudy.
CD4’weremonitoredattimeoneyearbeforetreatment(T-1)
andattheinitiationofthetreatment(T0)butthismonitoringcould
notbepursued.Weightandtotallymphocytenumberswere
moni-toredoneyearbeforetreatment(T-1),attime0(T0),3months(T3) and12months(T12)afterinitiationoffoodsupplementtreatment.
TreatmentwasstoppedatT12.
ThefoodsupplementwasapprovedbytheBelgianMinistryof
HealthonJune232009anddoesnotrequiretheapprovalofan
Ethicscommitteeforitsuse.
Results
Table1presentsresultsoffourteenpatientswhowereunder
conventional tri-therapy for about six months before addition
ofthefoodsupplementstothetreatment.Theirstateof illness
wasadvanced.Theregimenwasgivenaccordingtothemedical
imperatives defined by the treating physician. Three very ill
patientsunder tri-therapywho died either at thebeginning of
thetreatmentorwithinthefirstthreemonthsoftreatmentwere
omittedbecausetreatmentwithfoodsupplementsisalong-term
processthat couldnothave beeneffectiveattheirstageofthe
illness. Table2 showsthe resultsof46 AIDSpeople whowere
withoutspecificmedicaltreatment.Theirstateofillnesswasless severethanthepatientsundertri-therapy.Theresultsobtainedby
patientsundertri-therapyarepresentedatsomelengthbecause
theseresultsaremostinteresting.
Patientsundertri-therapy
P.P80,2LeidandP.I.andtri-therapy
Thefirstfourpatients(Nbs.1to4)wereinacriticalphysical conditionattimezero(T0)andreceivedthecombination[P.P80 +2Leid+P.I.+tri-therapy].Patient1wasa19-yearoldgirl.She weighed30kgatthebeginningofthetreatment(T0)andthe num-berofCD4cellshadcollapseddownto6.Evidently,shedidnot
benefitfromthetri-therapyandseemeddoomed.Inthreemonths
offoodsupplementsintake(T3),shegained15kgandher
num-beroftotallymphocytesincreasedfrom212to2915.Shewasin
goodhealthatT12,after12monthsoftheadditionalalternative treatment.
Thetri-therapyfollowedfromtimeT-1toT0appearedmore
beneficial topatients 2 to 4 becausethey maintained or even
slightlyincreasedinweightduringthattimeandhadretainedtheir
CD4andlymphocytelevels,albeitinverylownumbers.Thefood
supplements[P.P80+2Leid+P.I.andtri-therapy]causedadramatic gaininweight.Moreimportantly,therewasaspectacularincrease inthenumberoftheirtotallymphocytes.Allwereinasatisfactory stateofhealthafter12monthsoffoodsupplementsintake.
P.P.80,2Leidandtri-therapy
Patients5to8receivedonlyP.P.80+2Leid,inadditiontothe chemicals.Theywereinaverypoorstateofhealthbefore
start-ingfoodsupplementation:severedropsinCD4counts,exceptin
patient8,lowweightandlowlymphocytecount.Thetri-therapy
had proven ineffective. Afterthree monthsof foodsupplement
intake[P.P.80+2Leid+tri-therapy],lymphocytecountsincreased dramaticallyand3outof4patientshadasignificantgaininweight. Patient6alsogainedweightafter12monthsofalternativetherapy andallwereinasatisfactorystateofhealthatthatmoment(T12).
PP80andtri-therapy
Patients9to12receivedP.P.80inadditiontothetri-therapy. Patients9and10wereclinicallyspeakingina“stablestate”,with
relativelyhighCD4andlymphocytecountsattimeT0.Theweight
ofpatient9wasnotmeasuredbeforethestartofthestudy,but patient10hadastableweight.Bycontrast,patients11and12had
lowCD4countsdespitetri-therapyandhadlostweight.The
tri-therapyhadproveninefficient.Threemonthsaftertheystartedto taketheP.P.80foodsupplement,theirweightandtheirlymphocyte
countshadincreased.Aftertwelvemonthsoftreatmentwiththe
foodsupplement,theirhealthwasgood.
PP20,ParaImmunoandtri-therapy
Thisregimenwasgiventopatient13,a15-yearsoldgirl weigh-ing24kgatthetimeofheradmissionintheorphans’home,under
chemo-therapy but withdiarrhea, fever and pneumopathy and
havinglost
¾
ofherCD4’sduringchemicaltreatment,recoveredwhenP.P.20andP.I.wasgivenduringthreemonths.Theuseof
P.P.20insteadofP.P.80,a 4-foldlowerdoseinactiveprinciples,
resultedinasimilarweightgainandlymphocytecountrecovery
aswiththeprevious12patients,indicatingthatthislowerdosage wasjustasefficacious,atleastinateenager.
Patient14,undertri-therapyandreceivingonly2Leidasfood
supplementhadamoderateCD4cellcountbutwaslosingweight
beforejoining thestudy. Threemonthslater, shehad not only
Table1
FourteenAIDSpatientsunderconventionaltri-therapy.Thesepatientswereadvancedindisease,withonly#9and#10inaconservedstate.Forthisreason,theywere chemically-treatedbutthetreatmentsometimesworsenedtheircondition,shownbyalossinweight(e.g.#2,11,and14,orelseastatusquo:e.g.#5,#6,#10).Some underwentaseverelossofCD4duringthetreatment:e.g.#6,#7,#11,#12).Allgainedinlymphocytesnumbersduringfoodsupplementtreatment.
Patient Treatment Symptoms Weight ViralL CD4 Lymphocytes N◦ File Age Sex PP 2Leid P.I. T0 T-1 T0 T3 T12 T-1 T-1 T0 T0 T3 T12
1 LB42 19 F 80 yes yes 3,5,16,17 30 45 51 6 212 2915 3600 2 LA02 26 F 80 yes yes 3,6,5 55 50 62 69 120000 84 80 131 2760 3 LA49 34 F 80 yes yes verybad 46 51 62 70 125000 25 24 802 2615 4 LA50 36 F 80 yes yes verybad 56 62 72 83 125000 62 56 904 2464 5 LA18 20 F 80 yes 3,4,verybad 44 45 60 80 140000 34 2 151 1768 2916 6 LB41 20 M 80 yes 3,4,verybad 60 60 61 66 0 611 90 402 1915 2690 7 LA32 32 F 80 yes 3,4,verybad 44 46 56 60 98650 245 40 806 2900 3050 8 LA35 37 F 80 yes 3,4,verybad 47 55 72 79 150000 16 127 679 2464 9 LB33 12 F 80 Conserved 30 32 53 191 720 1920 4500 10 La15 21 F 80 conservedstate 59 60 65 76 20000 200 204 1504 2115 3915 11 LA06 30 F 80 2,3,5,6 46 40 53 67 120000 95 59 300 1584 3900 12 LA23 31 F 80 3,13,6,5, 55 50 66 75 135000 50 60 800 2160 3916 13 LA31 15 F 20 yes 3,6,5 24 26 33 40 0 435 100 802 2005 3050 14 LA07 23 F yes 6,3,11 45 40 52 64 10000 300 1401 2500 3900 ViralL:viralload.
Treatments:PP:paraaspido,20or80units;2Leid:cytokinesandnucleicacids;PI:royaljelly,propolis,pollen.Tritherapy:stavudine,lamivudine,nevirapine
Symptoms:1.congenital;2.adenopathy;3.diarrhea;4.dermatitis;5.pneumopathy;6.hyperthermy;7.Inappetence;8.shingles;9.otitis;10.pharyngitis;11.coughing;12. psychomotordelay;13.lymphadenopathy;14.candidiasis;15.vomiting;16.hepatitis;17.sarcoma.
doubled.These resultsarecomparable tothose observedinthe other13patients.
Patientswithouttri-therapy
Alargernumberofpatients(Table2:Nb15to60)receivedno tri-therapy.Thecliniciandefinedtheirhealthstatusattimezero byqualitativecriteria(conserved,good,badetc.)andtooknoteof opportunistinfections.ThelossofweightintheseAIDSpatients fromthemomentoftheiradmissionintheorphans’home(T-1)to timezero(T0),i.e.ayearlaterwhenfoodsupplementationstarted, isanobjectivesignoftheirevolution,asistheirviralloadandthe leveloftheirCD4’.Someofthem,especiallyyoungchildren,gained weightandincreasedthenumberofCD4whileotherssawadecline intheirCD4betweenT-1andT0,butonlytwoofthem(Nbs29and 41)hadCD4levelsbelowthe100markattimeT0.
Table2showsthat,irrespectiveoftheseverityoftheirillness
andtheworseningoftheirphysicalconditionduringtheyearthat
precededsupplementtreatment,allrecoveredandgainedweight
inadditiontoincreasingtheirtotallymphocytecount.Allwerein satisfactoryorgoodhealthattheendofthetwelve-monthcourse offood-treatment.Iftheeffectsofthistreatmentarevisibleafter
threemonthsinmostofthetreatedAIDSpeople,somehowever
neededmorethanthreemonthsoffoodsupplementintakebefore
showingclearevidenceofrecoveringhealth(Nbs21,27,56)even ifanincreaseinnumbersoftotallymphocytestookalreadyplace attimeT3.
Discussion
Duringthewholeyearoftreatmentwithnaturalproducts,we
deplorednocollateralsecondary effectsexcepttwo patientson
PP80,whosignalledanosebloodingthatdisappearedaftertheend ofthetreatment.Thisevidenceofinnocuitycorroboratestheresults obtainedinanimalstudies(Federlinetal.,2014).
Allappliedprotocolsprovedefficacious.Theuleinebooststhe synthesisofnitricoxide,theribonucleotidesstimulatethe multipli-cationoflymphocytesandtheParaImmuno(propolis,Royaljelly
andpollen)stimulatestheimmunesystem.Acombinationofall
threeworksbestinasmuchthateveryfacetoftheglobaltreatment wasexploited.However,thesingleapplicationofuleineto non-treatedpatients,beitatitshighestorlowestconcentration,yielded
remarkableresultsbyitself.Patients26,27and28,whoweregiven
PP80,showedasubstantialimprovementoftheircondition,ata
parwiththatobservedwhencompleteregimensweregiven.This
observationisvalidalsoforpatients47to51,whoreceivedPP20: theirconditionimprovedatthesamerateaspatientshandledwith
thetwoadditionalfoodsupplements.Thechemicallyunattended
patientsweregenerallyinabettercondition(i.e.said“conserved”)
thanthosewhowereundertreatment.Inthisgroup,patients9,
10, 11 and 12 gained noweight duringchemical treatmentor
elselostweight.Yet,PP80givenalonetotheminadditionto tri-therapyimprovedconsiderablytheirhealth.Anexplanationforthis remarkableeffectmaybefoundinthefactthatuleinealso inter-fereswiththeinflammatoryresponse(Nardinetal.,2008)and reg-ulatestheimmunesystem(Nardinetal.,2010).Thesubstance pos-sessesthuspropertiessharedbythetwootherregimenproposed. Onemustnoteherethatareductioninthenumberofpillstaken, andinthefrequencyoftheiradministration,mustbefavouredin allcasesnotonlyforreasonsofcostandburdenbutbecauseitplays aconsiderablepositiveroleonthepsychologyofthepatients.
Theprogressregisteredinitiallyontheclinicallevelbynearly allpatientsreceivingfoodsupplementswasconfirmedbya
signifi-cantincreaseoftotallymphocytesnumbersandweightincrease
from the 3th month on. Patients under tri-therapy progressed
mostobviouslyatthelevelofsecondaryeffectsand opportunis-ticinfections.In2006,attheendofthesupplementtreatment,we registerednocaseofdiarrhoea,nocaseofhyperthermia,nocase ofdermatoseorotheropportunisticillnesses.Wehadonlythree
deathsbypneumocytosis,meningitisandbacterialpneumopathy
butthesewereveryadvancedcasesatthebeginningofthe treat-ment.
The patientunder tri-therapy were given foodsupplements
when they showed no improvement and/or developed
oppor-tunisticdiseasesthatdidnotexistbeforethetreatmentwiththe anti-retrovirals.Atri-therapybasedonstavudine,lamivudineand nevirapineisaheavytreatmentthatoftencausessecondaryeffects inthepatientandcanbeafactorofhealthdegradationinmany treatedsubjects.Ourresultsindicatethattri-therapyworsensthe healthofsomeAIDSpatientsinsteadofimprovingit,while
tak-ingfoodsupplementsnotonlydoesnotcausethesamenegative
Table2
46patientsreceivingnostandardchemicaltherapy.Somepatientswerewithopportunisticdiseasesbutasizablenumberwereingoodstateor“conserved”.Allofthemsaw thenumberoftheirlymphocytesincreaseduringtreatment.
Patient Treatment Symptoms Weight ViralL. CD4 lymphocytes N◦ File Age Sex P.P. 2Leid P.I. T0 T-1 T0 T3 T12 T-1 T-1 T0 T0 T3 T12
15 LB52 15 F 80 yes yes 3,6,bad 45 30 35 45 0 610 924 1802 2720 16 LC03 17 F 80 yes yes conserved 44 54 60 1121 3915 17 LB64 19 F 80 yes yes conserved 58 69 73 1215 2280 18 LB60 20 F 80 yes yes goodstate 59 50 60 65 0 488 1320 3916 4200 19 LA21 23 F 80 yes yes goodstate 51 54 60 66 75000 95 420 1420 2850 3915 20 LA36 31 F 80 yes yes goodstate 60 65 73 81 0 500 318 1412 4080 21 LC02 35 F 80 yes yes 3,6,14,bad 40 42 76 902
22 LB23 20 F 80 yes conserved 60 65 70 74 6000 296 315 1390 2500 3920 23 LA20 21 F 80 yes +/-’good 48 52 58 64 122000 50 105 1240 1539 2620 24 LA39 35 F 80 yes conserved 40 45 65 75 120000 25 312 1219 2900 3620 25 LC04 M 80 yes 3,6,17 50 60 65 502 2108 26 LB30 20 F 80 conserved 35 50 61 66 10000 205 310 1311 2530 3910 27 LB19 20 F 80 goodstate 60 65 66 75 0 391 320 1301 1320 3900 28 LA33 30 F 80 conserved 46 50 60 65 100050 250 360 1250 2500 3200 29 LA13 3 F 20 yes yes 3,6,5,12 6 7 12 18 130000 80 60 1201 2100 4610 30 LA45 3 F 20 yes yes goodstate 7 12 17 23 125000 25 391 1210 4608 5105 31 LA26 9 M 20 yes yes goodstate 14 16 20 36 0 420 402 1240 2100 4112 32 LA08 10 F 20 yes yes 6,3,7,4,9 21 21 30 46 6000 300 277 1314 3915 4904 33 LA44 10 F 20 yes yes goodstate 14 20 30 37 125000 85 420 1311 3016 3620 34 LA05 12 F 20 yes yes 1,6,3,8 28 29 35 51 12000 250 277 1360 3005 4915 35 LA09 12 F 20 yes yes 1,9,4 25 25 38 45 0 350 244 1901 2615 3916 36 LB53 13 M 20 yes yes verybad 13 20 25 612 3360 4600 37 LA04 14 F 20 yes yes 5,8 31 28 35 43 .50.000 140 130 1211 2900 4190 38 LA40 14 F 20 yes yes goodstate 16 20 30 46 53500 250 305 1229 3050 4215 39 LA01 11 M 20 yes 2,3,4,5,10 26 24 30 35 80500 120 100 711 2110 40 LB46 21 F 20 yes conserved 59 65 75 226 1247 3024 4500 41 LA27 4 M 20 yes good 11 11 15 21 0 500 305 1321 1910 2915 43 LA41 10 M 20 yes good 16 20 26 40 25000 350 315 1319 3600 4250 44 LA17 11 M 20 yes +/-’good 15 20 25 35 130000 100 60 405 1120 2905 45 LA30 13 M 20 yes conserved 19 21 31 36 0 600 372 1120 2820 3912 46 LA03 15 F 20 yes 3,7 45 41 50 60 110000 91 300 1707 2910 4210 47 LA43 9 F 20 good 13 20 25 36 15000 350 416 1620 2600 3950 48 LA29 11 M 20 conserved 24 25 30 36 0 480 450 1200 2912 4045 49 LB35 12 F 20 conserved 19 22 29 35 0 412 383 827 1460 2920 50 LB32 13 F 20 Conserved 30 32 40 45 5000 490 259 561 1768 2900 51 LB13 19 M 20 goodstate 60 62 75 79 0 526 620 1302 4901 52 LA10 21 F yes yes 60 55 61 72 30000 200 1500 2500 4600 53 LB04 3 F yes 3 12 12 15 26 0 710 1400 3500 54 LB02 4 F yes 3,6,delayedgrowth 12 13 18 24 0 420 1830 2930 55 LB17 5 F yes 3,15,14 18 18 25 30 0 518 1150 3900 4600 56 LB08 7 F yes 6,14 18 18 25 37 0 520 1200 4500 57 LB21 21 F yes 3,6,7 60 42 45 50 0 620 1950 2500 3600 58 LA25 25 F yes 3,4,14 44 45 60 65 10000 360 1300 3900 4115 59 LA22 26 yes conservedstate 55 50 61 67 8000 300 1200 2700 4105 60 LA47 33 F yes 3,6,5tuberc. 52 51 61 71 5500 350 1250 2950 3920 LA38 33 M yes conservedstate 46 46 55 66 60000 300 1220 2915 2400 ViralL:viralload.
Treatments:PP:paraaspido,20or80units;2Leid:cytokinesandnucleicacids;PI:royaljelly,propolis,pollen.Tritherapy:stavudine,lamivudine,nevirapine
Symptoms:1.congenital;2.adenopathy;3.diarrhea;4.dermatitis;5.pneumopathy;6.hyperthermy;7.Inappetence;8.shingles;9.otitis;10.pharyngitis;11.coughing;12. psychomotordelay;13.lymphadenopathy;14.candidiasis;15.vomiting;16.hepatitis;17.sarcoma.
Authors’contributions
DMcontributed in thefabrication of the foodsupplements, deliveryandfollowup;andRManalysedtherawdata,exploited them and wrote the manuscript. Both authors have read and approvedthepaperforsubmission.
Conflictofinterest
Allauthorshavenonetodeclare
Acknowledgment
DoctorChristianBouillonand ChloeNdayikundaprovidedus withthefilesofthepatients.
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