telling the truth – my experiences with lyme borreliosis – bózsik, béla pál, m. d. lyme...
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Telling The TruthTelling The Truth– – my experiences with Lyme my experiences with Lyme
borreliosisborreliosis – –
Bózsik, Béla Pál, M. D.Bózsik, Béla Pál, M. D.Lyme Borreliosis Foundation,Lyme Borreliosis Foundation,
hUNGARY hUNGARY
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By Mel Gibson
think of the real truththink of the real truth
Dialogue on Dialogue on veritasveritas from Claudia & pilatusfrom Claudia & pilatus
What is the veritas, claudia?What is the veritas, claudia?Have You heard recognized if it would be said? Yes, I do!Have You heard recognized if it would be said? Yes, I do!How?-could you tell me?How?-could you tell me?If you have not heard it nobody could tell YouIf you have not heard it nobody could tell You
VERITAS?!–My verity is THAT any way it will bloodshed here!VERITAS?!–My verity is THAT any way it will bloodshed here!VERITAS?!–If it happens MY BLOOD WILL SHED BY KAISER!VERITAS?!–If it happens MY BLOOD WILL SHED BY KAISER!
If you have not heard the verityIf you have not heard the verity nobody could tell Younobody could tell You
The PassionThe PassionsequenceS ON THE TRUTHsequenceS ON THE TRUTH
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WELL,WELL, What is the TRUTH? What is the TRUTH?
OMAN:OMAN:
ANAN::
It could be personally It could be personally sensesense..
It depends on theIt depends on the principalitiesprincipalities..
It is better to sacrifice It is better to sacrifice oneonefor the people. John for the people. John 18,1418,14
ANHEDRIN:ANHEDRIN:
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Y opinion:
What is the TRUTH What is the TRUTH in Lyme Borreliosis? in Lyme Borreliosis?
Lyme borreliosis
ENDEMIC both in the World & in Hungary is
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Was estimated to affectWas estimated to affect
as MUCH as as MUCH as 10 %10 % of the populationof the population
at leastat leastBózsik, Lancet 2004Bózsik, Lancet 2004
Regarding to diagnostic and differential diagnostic difficulties there is no exact epidemiological data
Due to therapeutic problems and the long persistence of Lyme borreliosis the patients are accumulated in the population.
Bózsik, Vienna 1985Bózsik, Vienna 1985
This endemic This endemic Lyme borreliosisLyme borreliosis
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This Estimation based on bothThis Estimation based on both thethe possibility of tick’s co-infectionspossibility of tick’s co-infections& the cumulation of lyme borreliosis& the cumulation of lyme borreliosis
1: 101: 10
Lyme borreliosis incidence = ––––––––––– Lyme borreliosis incidence = ––––––––––– 200 TBE200 TBE_cases/years_cases/years
1: 1 000 1: 1 000
BacteriumBacterium
LB = TickLB = Tick’s infectivity’s infectivity rate with –––––––––––– rate with –––––––––––– TBE TBE_cases_cases
VirusVirus
Twenty thousends new cases yearly in Hungary
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What thisWhat thismeans & causes?means & causes?
No definiteNo definite Control popu- Control popu-
lationlationDiagnosis Diagnosis either ineither in laboratory laboratory oror clinical clinical Hampered Hampered by theby the protean symptomsprotean symptomsProceedings against Proceedings against MD-sMD-s
VainVain Fiery debates Fiery debates
in the society in the society . Heavy & long suffer-ing of Heavy & long suffer-ing of
patientspatients Decreased producti-vity of Decreased producti-vity of
the society.the society. Increased claims Increased claims
to rto rx . x . Increased Increased
medical costs medical costs . .
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ARE the FOLLOWINGARE the FOLLOWING The right answerS? The right answerS?
„„There is no such disease, There is no such disease,
if it could exist at all.”if it could exist at all.”
„„You surely not have this!”You surely not have this!”
Doctors SAY in all countries of the Doctors SAY in all countries of the world.world.
Could it really answeredCould it really answered
by serology? - I have to try it .by serology? - I have to try it .
Carl Brenner Science, 1992
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OUR PRACTICE WAS OUR PRACTICE WAS SINCE 1984 IN HUNGARYSINCE 1984 IN HUNGARY
0
4000
8000
12000
16000
20000
ESETSZÁM 139 715 2437 7071 11617 12283 13983 14207 13811 18698 14971
Vizsgálatszám 139 857 2689 7793 12752 13299 15003 15101 14596 19468 15863
POZITIV 106 420 1138 2958 3024 4549 3823 2369 3671 5013 3206
POZITIVITÁS % 76,3 49 42,3 38 23,7 34,2 25,5 15,7 25,2 25,8 20,2
1987-Vienna
1988-Vienna
89. 90. 91. 92. 93. 94. 95. 96. 97.
SEROLOGY FOR LYME BORRELIOSISJOHAN BÉLA NATIONAL INSTITUTE OF HYGIENE,
LABORATORY OF SEROLOGY 1987 - 1997109 932 CASES117 560 PHA-investigation 30 277 POSITIVE (25.8%) 1 349 ELISA-investigation 715 POSITIVE (53.0%)
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Lyme borreliosis is endemic in Lyme borreliosis is endemic in hungary hungary
determinations at the nih, hungarydeterminations at the nih, hungary
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Dinamic changing in serologyDinamic changing in serologyproduction of antibodiesproduction of antibodies
Summarized DiagramSummarized Diagram
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Dinamics of Antibody production.Dinamics of Antibody production.It binds to every antigens It binds to every antigens of of borrelia burgdorferi s.l.borrelia burgdorferi s.l.
Effects of reinfection(s)Effects of reinfection(s)
Effects Effects onon IgMIgM production production
Persisting IgPersisting Igss & lb & lb
Eff
ects
of
ab
xEff
ects
of
ab
x
iigggg
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Dinamic changing both in Dinamic changing both in serology & serology & production of antibodiesproduction of antibodies
Th
e p
rob
lem
is in
dete
rmin
ing
Th
e p
rob
lem
is in
dete
rmin
ing
th
e r
igh
t cu
t-off
level
the r
igh
t cu
t-off
level
infectioninfection
ecmecm
sec.sec.ecmecm
Changing in antigenecityCauses IgM elevations,
just as If it would be new infections
Decapitated, inhibited immune response
cut-off level?
Persistent IgPersisting lb
Spirochetaemia during the whole pathogenesis
TH
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Is it possible to describe all of these Is it possible to describe all of these changes or changes or
diagnose Lyme borreliosis diagnose Lyme borreliosis
using any methodusing any method??
The question arised :The question arised :
No, that is the answer of course.No, that is the answer of course.
Determining the cut-off level Determining the cut-off level is also disputable is also disputable
as it is hard to determineas it is hard to determine
with with oneone investigation investigation
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Lyme borreliosis reinfectiosaLyme borreliosis reinfectiosa
could only be revealed with early serologic testing, as
immune response of the body is slow & lasting: therefore existence of the antibody in the presence of ECM could be evaluated as reinfection.
can Lyme borreliosis be can Lyme borreliosis be over/UNDER-diagnosed over/UNDER-diagnosed
Mis-diagnosis Mis-diagnosis is the only possibility!is the only possibility!
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Tail-cutting helps to evaluateTail-cutting helps to evaluate the results of passive haemagglutinationthe results of passive haemagglutination
The evaluation is based on 120,000 samples & 150,000 determinations since 1989.
The cut-off levelcan be calculated
from the resultsalone
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Tail-cutting Tail-cutting for evaluating serologyfor evaluating serology
Reactivity of samples wereReactivity of samples wereDetermined by the dilution & cross(1-4)Determined by the dilution & cross(1-4)-Number versus 2/base log of dilution--Number versus 2/base log of dilution-
Highly reactive sera Highly reactive sera are few to delineateare few to delineate
Sera with low reactivitySera with low reactivity
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There are two populationsThere are two populationsSmall Positive(red)Small Positive(red)
Large Negative(blue)Large Negative(blue)They are Clearly dividedThey are Clearly divided
But missed But missed diagnosis of 30500 patientsdiagnosis of 30500 patients
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Critical fields Critical fields to evaluationto evaluation
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0
0,02
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5 5,5 6 6,5 7 7,5 8 8,5 9 9,5 10
0
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5 5,5 6 6,5 7 7,5 8 8,5 9 9,5 10
0
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5 5,5 6 6,5 7 7,5 8 8,5 9 9,5 10
0
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5 5,5 6 6,5 7 7,5 8 8,5 9 9,5 10
0
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5 5,5 6 6,5 7 7,5 8 8,5 9 9,5 10
The parts of The lastsNormal distribution curves
for determining the cut-off level
Critical fields Critical fields to evaluationto evaluation
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7,6
7,8
8
8,2
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8,6
8,8
9
9,2
3132333435363738390
2
4
6
8
10
12
(log2Dilution) 2nd failure in % 1st failure in %
Plato areaWhere are potentially the less
missed cases
A Plato describedBy the errors of the curves
They Determined the cut-off levels
The cut-off level 1:200 dilution 3+ reactivity
Causing the smallest in both failures:Less the missed patients as seronegative: under 1,5% or it gives the less seropositivity to healthy:
under 5,5%missed 7,0%.
Although This is excellent result, it needs further evaluation For the benefit of the lyme-patients
10% missed
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Which method could became Which method could became
the golden standard?the golden standard?
Or
Som
eO
r S
om
eFu
rth
er
new
meth
od
sFu
rth
er
new
meth
od
s
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Lyme borreliosis seronegativaLyme borreliosis seronegativain relation to different methodsin relation to different methods
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As you have seenthere is a great difference between
biological & biostatistical error
Although 1.5 % first order and 5.5 % second order error are excellent results for any laboratory it is astonishing that the developments of science give possibility to determine further 60 % or more cases as people, who are in bed need of treatment.
This result is neither over-diagnosis, nor under-estimation, especially not fals/untrue determinations
they really belong to both the development of methodology and the epidemic character of Lyme borreliosis.
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I think of I think of many tenmany ten--thousands of peoplethousands of people
with suspected symptoms with suspected symptoms stated by their physicians stated by their physicians
Who could not get Who could not get the right answer or the right answer or
the definitive diagnosis.the definitive diagnosis.
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The BLUE COLUMNS The BLUE COLUMNS related to the missed proportions of related to the missed proportions of
patientspatients
theore
tical g
rap
h!
theore
tical g
rap
h!
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4/1802,2%
2/1801,1%negative
3/1801,7%
171/18095%Positive
negativePositive
Western-blot
There was not any reactivity in Master’s disease, in which B. burgd. sensu lato were isolated with substrain determination.
Masters,E–Philipp,M:Masters,E–Philipp,M: C6 Lyme peptide ELISA serosurvey of Missouri patients.Abstracts&Presentations at the IX Intl.Conf. on Lyme borreliosis &
Abstract at The Clinical side of Lyme Disease, N.Y. Augustus 18-22., 2002.
What is on C6-ELISA?What is on C6-ELISA?C6 ELISA & Wb
were identical in 97% at CDC comparing 180 samples
The molecular biologyThe molecular biology of the VlsEof the VlsE-antigen-antigen
is well known!is well known!
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It also teaches us It also teaches us that for evaluation that for evaluation
it should not be forgottenit should not be forgotten:: Lyme borreliosis Lyme borreliosis
Is a worldwide epidemicIs a worldwide epidemicWith countless ManifestationsWith countless Manifestations
The high incidence & The high incidence & especially extra high prevalenceespecially extra high prevalence
Is characteristic for Lyme borreliosisIs characteristic for Lyme borreliosis
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“The Committee recognizes that the current state of laboratory testing for Lyme disease is very poor. The situation has led many people to be misdiagnosed and delayed proper treatment. The vaccine clinical trial has documented that more that one third ( 36 percent ) of the people with Lyme disease did not test positive on the most sophisticated tests available. The ramifications of this deficit in terms of unnecessary pain, suffering and cost are staggering. The Committee directs CDC to work closely with the Food and Drug Administration to develop an unequivocal test for Lyme disease.( Public Law 107-116 Signed by President Bush 1/10/02 Departments of Labor, Health, and Human Services, and Education, and Related Agencies Appropriations Act 2002)
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Is there Is there any solution any solution
of the diagnostic problemsof the diagnostic problems
Nearly one hundred yearsNearly one hundred yearsColeman, 1909Coleman, 1909
There has been a simple & good one!There has been a simple & good one!
It is possible It is possible To detect spirochetesTo detect spirochetes
With dark-field microscopyWith dark-field microscopy&&
since Nearly that time at leastsince Nearly that time at leastIt is debatedIt is debated
Because of the so-calledBecause of the so-calledMyelin figures, whichMyelin figures, whichLook like spirochetesLook like spirochetes
( pseudo-spirochetes )( pseudo-spirochetes )
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The largest energies The largest energies
in the naturein the nature
Among them Among them Is Is
the bloodthe blood
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There is direct connection There is direct connection between the body & the tickbetween the body & the tick
timetimeIs importantIs important
For the infectivity, becauseFor the infectivity, becauseTicks infect us Either with salivaTicks infect us Either with saliva
Or with regurgitation.Or with regurgitation.
this connection, this connection, makes possible for spirochetesmakes possible for spirochetes
to circulateto circulate&&
To reach the hidden places, too.To reach the hidden places, too.
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therefore, there is therefore, there is
no useno use to be distinguished to be distinguished
stagesstages for a generalized ailment,for a generalized ailment,
as it will be forgotten: as it will be forgotten:
Lyme borreliosis Lyme borreliosis damages the whole bodydamages the whole body
from the first moment.from the first moment.
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LLyme borreliosisyme borreliosis seronegativaseronegativa
one third ( 36 percent ) of the people with Lyme disease did not test positive
Public Law 107-116 Signed by President Bush 1/10/02
Nearly the same proportions Nearly the same proportions of lyme-patients of lyme-patients
were provedwere provedAs Lyme borreliosis seronegativa As Lyme borreliosis seronegativa in my practice with DF-microscopyin my practice with DF-microscopy
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My practice withMy practice withdarkfielddarkfield--microscopymicroscopy
was proven with realwas proven with real-time pcr-time pcr
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According to my practice According to my practice since 1986 since 1986
borrelia burgdorferi s. l. borrelia burgdorferi s. l. Could be demonstrated & studied with dark-Could be demonstrated & studied with dark-
field microscopy field microscopy during the whole pathogenesis.during the whole pathogenesis.
Besides clinical signs/symptoms Besides clinical signs/symptoms && serology serology it was proved it was proved
immunocytologically immunocytologically with kindly donated MCwith kindly donated MC
of Barbour & Steiner of CDC () of Barbour & Steiner of CDC () specific staining of rutenium red andspecific staining of rutenium red and
electron microscopy with negative staining.electron microscopy with negative staining.
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DualDurDualDur ®® reagent & methodreagent & method help to overcome difficulties in diagnosis of help to overcome difficulties in diagnosis of
Lyme borreliosis seronegativaLyme borreliosis seronegativa
first
second
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Dividing Spirochete beside two
thrombocytes
Division of Spirochete happens
With Central concentration & splitting
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immuncytology (1990)immuncytology (1990): : Indirect immuno-fluorescence with Monoclonals anti-OspAIndirect immuno-fluorescence with Monoclonals anti-OspA és anti-és anti-flagellaris antitestekkel(prof.Barbour)flagellaris antitestekkel(prof.Barbour)
anti-anti-flflagellarisagellaris
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TheoreticalTheoretical – Length: 15-30 μm, Width: 0,15-0,30 μm, Flagellum: 7--20This oneThis one – Length ~15 μm, Width: 0,15-0,30 μm, NO Flagellum
Kari Hovind-Hougen, 1976-1986
Morphology of BorreliaeMorphology of Borreliae
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In 1909 & detailed in 1913:In 1909 & detailed in 1913:a new spirocheta was found a new spirocheta was found
in Human blood at graves’ diseasein Human blood at graves’ diseaseChambers,H-Royal Free Hospital
BMJ, 1909 & Lancet 1913,1:1728
„Diagrammatic representation of the Spirochetae seen in normal serumwhen examined by dark ground illumination”
(Some of these )
HOMOLOGIES between proteins of Borrelia burgdorferi & Thyroid AUTOANTIGENS
THYROID 2004, 14:86
great honourgreat honourto helen chambersto helen chambers
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Borrelia burgdorferi s.s.Borrelia burgdorferi s.s. -Morphology in the friendly -Morphology in the friendly
cultivation media-cultivation media-Aberer-Duray, 1991
Meduza-like (paired-crossed-)
Double membranesVibrio-likeBipolar Taper
Treponema-like forms Paired Cells
Leptospira-likes Overlap/joinedElbow
UnipolarTaper
Before dividing? (Central splitting!)
End-joined Spirochetes
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various forms of various forms of Borrelia Borrelia burgd.s.l.burgd.s.l.
in blood usingin blood using DualDurDualDur®®After the basic study of Aberer-Duray, 1991
Double membranesVibrio-likeBipolar Taper
Treponema-like forms Paired Cells
Leptospira-likes Overlap/joinedElbow
UnipolarTaper
End-joined Spirochetes
Before dividing? (Central splitting!)
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Patients’ Blood sample (27/72/105; 72/14/14) Phosphorwolframic-acid (1%) negativ staining, 1000x
Vesiculum, extrabacterialis proteinVesiculum, extrabacterialis protein bborrelia burgd.s.l.orrelia burgd.s.l.
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A. KERSTEN, C. POITSCHEK, S. RAUCH, AND E. ABERER: Effects of Penicillin,
Cettriaxone, and Doxycycline on Morphology of Borrelia burgdorferi
Antimicrob agent s chemother 1995,39:1127-33
Blebbing Blebbing in medical literaturein medical literatureDuring cultivation & under abxDuring cultivation & under abx
Ghost-likereorganizedmembrans
blebs
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Extrabacterialis clouds in the layer of the Spirochete.Flagellum not visible.
Periplasmatic space
– Blood sample, Lyme-patient; Magnification 80.000x.
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Borrelia burgdorferi s.l.Borrelia burgdorferi s.l. Immuncytologic experiments in 2003Immuncytologic experiments in 2003
Patient blood sample (GD-222)CDC, Monoclonalis antitest OspA+Avidin-Biotin Immunperoxidáz reaction,
Original magnification 100x
Control
Blood sample Blood sample
Blood
samples
Blood sample
Blood sample
Blood sample
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Morphology ofMorphology of Borrelia burgdorferi Borrelia burgdorferi s.s.s.s. During cultivation - for controlDuring cultivation - for control
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Patient blood sample GD-223MC-OspA-AvidinBiotinPO
GD-223MC-OspA-AvidinBiotinPO
Shed ding for both defe nce & diseaseAs they are immu nocomplexes.
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There
were
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There
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ith
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eliosis
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h L
ym
e b
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eliosis
ch
ron
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Was h
eale
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as h
eale
d
DarkfieldsDarkfieldsSame prep.Same prep.
stainedstained
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borreliae could be detected by specific borreliae could be detected by specific staining staining of of the outer layerthe outer layer with ruteniumwith rutenium--
red (dark-gray)red (dark-gray)
Please admire extra-bacterial materialPlease admire extra-bacterial materialaround the spirochetes (yellow)around the spirochetes (yellow)
Pati
en
tP
ati
en
t w
ith
Lym
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eliosis
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eliosis
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Was h
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Acid Mucopolysaccharide
layer (dark gray)
protoplasmaprotoplasma
protoplasmaprotoplasma
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ClosingClosing remarksremarks
Definitive diagnosis Definitive diagnosis for Lyme borreliosis for Lyme borreliosis
could be set upcould be set up with laboratory methods.with laboratory methods.
indication indication of the treatmentof the treatmentneeds consultation,needs consultation,
that should be followedthat should be followedby control determinations by control determinations
for yearsfor years..
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we should have to unit we should have to unit our knowledgeour knowledge
„„As our enemy, the devil” As our enemy, the devil” is circulating &is circulating &
Wanted to devour us....Wanted to devour us....
11stst Letter of Peter 5 Letter of Peter 5ththchapter verse 8chapter verse 8