s.s.j hepatoprotective protocol.docx

8/9/2019 s.s.j hepatoprotective protocol.docx

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Albino research &training institute

Hepatoprotective activit of methanolic extract of casearia eliptica leaf extract in rats

ABSTRACT:

Liver is the largest organ in the body which serves as a gland also. It plays an important role in

the maintenance of internal environment through its multiple and diverse functions. Any damage

to the liver or impairment of its functions leads to injurious effects. Liver diseases (like jaundice)

are the common ailments affecting mankind, though no remedy is available in allopathy at

present. ain is defined as neuralgia, an unpleasant sensory e!perience associated with tissue

damage. Inflammation is a common complaint in most patients suffering from disease conditions

.Inflammation is a normal protective response to tissue injury caused by physical trauma,

no!ious chemical or microbial agents. In the recent past years many medicinal plants are

screened for their hepatoprotective as well as analgesic and anti inflammatory activity and "uite

a few of them are already successful in entering the market, #ence the present study is planned

to find out the hepatoprotective and analgesic and anti inflammatory activity of caseria eliptica

leaf using drug induced hepatoto!icity paracetamol,ccl$ model respectively.

Keyor!s:casearia eliptica leaf, paracetamol,ccl4, hepatoprotective activity

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"#TR$%CT"$#:

Hepatotoxicity:

Liver is the largest organ in the body which serves as a gland also. It is wedge shaped and

weighs between %&'' to %''gms. It lies in the right upper "uadrant of the abdominal cavity,

beneath the diaphragm and also attached to it. It is divided into four lobes (uyton *.Arther,

%+-).It plays an important role in the maintenance of internal environment through its multiple

and diverse functions (alwar ., %++).

he liver stores a multitude of substances, including glucose (in the form of glycogen),

vitamin A (%/& years0 supply), vitamin 1 (%/$ months0 supply), vitamin 2%& (%/3 years0

supply), iron, and copper.

he liver is responsible for immunological effects4the reticuloendothelial system of the

liver contains many immunologically active cells, acting as a 0sieve0 for antigens carried to it via

theportal system.

he liver produces albumin, the major osmolarcomponent ofblood serum.

he liver synthesi5es angiotensinogen, a hormone that is responsible for raising the blood

pressurewhen activated byrenin,an en5yme that is released when the kidneysenses low blood

pressure.

Any damage to the liver or impairment of its functions leads to injurious effects. Liver

diseases (like jaundice) are the common ailments affecting mankind, though no remedy is

available in allopathy at present (2hat 1.Arun, et.al, %++-).

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http://en.wikipedia.org/wiki/Glycogenhttp://en.wikipedia.org/wiki/Vitamin_Ahttp://en.wikipedia.org/wiki/Vitamin_Dhttp://en.wikipedia.org/wiki/Vitamin_B12http://en.wikipedia.org/wiki/Ironhttp://en.wikipedia.org/wiki/Copperhttp://en.wikipedia.org/wiki/Reticuloendothelial_systemhttp://en.wikipedia.org/wiki/Hepatic_portal_systemhttp://en.wikipedia.org/wiki/Hepatic_portal_systemhttp://en.wikipedia.org/wiki/Albuminhttp://en.wikipedia.org/wiki/Osmolarhttp://en.wikipedia.org/wiki/Blood_plasmahttp://en.wikipedia.org/wiki/Angiotensinogenhttp://en.wikipedia.org/wiki/Angiotensinogenhttp://en.wikipedia.org/wiki/Blood_pressurehttp://en.wikipedia.org/wiki/Blood_pressurehttp://en.wikipedia.org/wiki/Reninhttp://en.wikipedia.org/wiki/Reninhttp://en.wikipedia.org/wiki/Reninhttp://en.wikipedia.org/wiki/Juxtaglomerular_apparatushttp://en.wikipedia.org/wiki/Juxtaglomerular_apparatushttp://en.wikipedia.org/wiki/Hypotensionhttp://en.wikipedia.org/wiki/Hypotensionhttp://en.wikipedia.org/wiki/Vitamin_Ahttp://en.wikipedia.org/wiki/Vitamin_Dhttp://en.wikipedia.org/wiki/Vitamin_B12http://en.wikipedia.org/wiki/Ironhttp://en.wikipedia.org/wiki/Copperhttp://en.wikipedia.org/wiki/Reticuloendothelial_systemhttp://en.wikipedia.org/wiki/Hepatic_portal_systemhttp://en.wikipedia.org/wiki/Albuminhttp://en.wikipedia.org/wiki/Osmolarhttp://en.wikipedia.org/wiki/Blood_plasmahttp://en.wikipedia.org/wiki/Angiotensinogenhttp://en.wikipedia.org/wiki/Blood_pressurehttp://en.wikipedia.org/wiki/Blood_pressurehttp://en.wikipedia.org/wiki/Reninhttp://en.wikipedia.org/wiki/Juxtaglomerular_apparatushttp://en.wikipedia.org/wiki/Hypotensionhttp://en.wikipedia.org/wiki/Hypotensionhttp://en.wikipedia.org/wiki/Glycogen


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'"T(RATR( R()"(*:

%. 6.7arthikeyan8 hepatoprotective activity ethanolic e!tract spermacoce hispida linn against

*cl$induced hepatoto!icity on albino wistar rats,

9anuary : &'%' ;


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subjected to antio!idant study using the &, &:diphenyl:%:picrylhydra5yl (1#) radical

scavenging assay with the total phenolic content (*) also determined6?2 e!erts potential

hepatoprotective activity that could be partly attributed to its antio!idant activity and high

phenolic content and thus warrants further investigation.

3.I.@. Ashousha: Antio!idant activity and hepatoprotective effect of pomegranate peel and whey

powders in rats,ajendra =. 1ash, 6ohammed #abibuddin, 1ev 2. 2aruah8#epatoprotective activity of

e!tracted anthocyanins fraction of red radish (Raphanus sativusL) on albino rats

9 ?!p Integr 6ed. &'%3G 3(%)8 $3:'

doi8 %'.$;jeim.&&'%&.or.'$-

Ee investigated hepatoprotective and antio!idant activity of e!tracted anthocyanins fraction of

red radish for the first time.

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http://www.sciencedirect.com/science/article/pii/S0570178313000067http://www.sciencedirect.com/science/article/pii/S0570178313000067http://www.sciencedirect.com/science/journal/05701783/58/1http://www.scopemed.org/?jid=4http://www.scopemed.org/?jid=4&iid=2013-3-1.000http://dx.doi.org/10.5455/jeim.220812.or.046http://www.sciencedirect.com/science/article/pii/S0570178313000067http://www.sciencedirect.com/science/article/pii/S0570178313000067#aff1http://www.sciencedirect.com/science/journal/05701783/58/1http://www.scopemed.org/?jid=4http://www.scopemed.org/?jid=4&iid=2013-3-1.000http://dx.doi.org/10.5455/jeim.220812.or.046


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Anthocyanins fraction of red radish (Raphanus sativusG AB>@) was selectively e!tracted by

employment of polymeric ion:e!change resin. AB>@ was evaluated for antio!idant and

hepatoprotective activity against carbon tetrachloride (**l$):induced hepatoto!icity in rats. his

study demonstrates the antio!idant and hepatoprotective activity of anthocyanins fraction

isolated from Raphanus sativusand thus scientifically supports the usage of it as food colorantand also justifies the use of the crude e!tracts of radish to treat various liver ailments in Indian

folk medicine.

.

-. @udipta 1as8 ?valuation of Anti:Inflammatory Activity of *lerodendron infortunatum Linn.

?!tract in >ats,lobal 9ournal of harmacology $ (%)8 $:', &'%' I@@= %++&:''H I1@I

ublications, &'%'

Inflammation is a response of vasculari5ed living tissue to the local injury. he severe side

effects of steroidal and non:steroidal anti:inflammatory drugs evoked us to search for new anti:

inflammatory drugs from the indigenous source. he methanol e!tract of leaves of *lerodendron

infortunatum Linn. (6?*I) was evaluated for anti:inflammatory activity against the carrageenan,

histamine and de!tran induced rat paw edema..

H. Akash A 7hobragade%F, @adi" 2 atel&, >upesh > ophale%, o!ithromycin and ?rythromycin, Alone and in

*ombination with Ibuprofen,I@> 9ournal of harmacy


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. =irmala 6F, irija 7, Lakshman 7, 1ivya 8#epatoprotective activity of 6usa paradisiaca

on e!perimental animal models8 Asian acific 9ournal of ropical 2iomedicine. 9ournal

homepage8www.elsevier.com;locate;apjtb

#epatoprotective activity of alcoholic and a"ueous e!tracts of stem of 6usa paradisiaca

was demonstrated by using **l$ and paracetemol induced hepatoto!ic models.

+. @u5y 6 @alama%, 6ahmood Ameen Abdulla%F, Ahmed @ Al>ashdi%, @almah Ismail&, @alim @

Alkiyumi and @hahram 8#epatoprotective effect of ethanolic e!tract of Curcuma longaon

thioacetamide induced liver

cirrhosis in rats8

he hepatoprotective effect of *L>? was measured in a rat model of thioacetamide:induced

liver cirrhosis over weeks. #epatic cytochrome $' &?% and serum levels of B:K% and

=B: were evaluated. !idative stress was measured by malondialdehyde, urinary :

hydro!yguanosine and nitrotyrosine levels.

%'. 1hanabal @F%, >ahul 9ain%, riyanka 1warampudi L%, 6uruganantham =%, >aghu8

#epatoprotective activity of Santolina chamaecyparissus Linn against 1:alactosamine Induced

#epatoto!icityin >ats

@&G harmacognosy *ommunications www.phcogcommn.org


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A"+ A#% $B,(CT")(S

he aim of the study is to evaluate the #epatoprotective,Analgesic leaf e!tract of casearia

elliptica lea) used in e!perimental animal models.

%.o prepare whole plant e!tract of casearia elliptica lea)

&.o evaluate the hepatoprotectve activity

*cl$ induced hepatoto!icity

aracetamol induced hepatoto!icity

hioacetamide induced hepatoto!icity

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lant profile 8

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Scientific classification

7ingdom8 lantae

(unranked)8 Angiosperms

(unranked)8 ?udicots

(unranked)8 >osids

rder8 6alpighiales

Bamily8 @alicaceae

ribe8 @amydeaeO%P

enus8 Casearia

9ac".O&P
http://en.wikipedia.org/wiki/Biological_classificationhttp://en.wikipedia.org/wiki/Planthttp://en.wikipedia.org/wiki/Angiospermshttp://en.wikipedia.org/wiki/Eudicotshttp://en.wikipedia.org/wiki/Rosidshttp://en.wikipedia.org/wiki/Malpighialeshttp://en.wikipedia.org/wiki/Salicaceaehttp://en.wikipedia.org/w/index.php?title=Samydeae&action=edit&redlink=1http://en.wikipedia.org/w/index.php?title=Samydeae&action=edit&redlink=1http://en.wikipedia.org/wiki/Casearia#cite_note-1http://en.wikipedia.org/wiki/Jacq.http://en.wikipedia.org/wiki/Jacq.http://en.wikipedia.org/wiki/Jacq.http://en.wikipedia.org/wiki/Casearia#cite_note-GRIN-2http://en.wikipedia.org/wiki/Biological_classificationhttp://en.wikipedia.org/wiki/Planthttp://en.wikipedia.org/wiki/Angiospermshttp://en.wikipedia.org/wiki/Eudicotshttp://en.wikipedia.org/wiki/Rosidshttp://en.wikipedia.org/wiki/Malpighialeshttp://en.wikipedia.org/wiki/Salicaceaehttp://en.wikipedia.org/w/index.php?title=Samydeae&action=edit&redlink=1http://en.wikipedia.org/wiki/Casearia#cite_note-1http://en.wikipedia.org/wiki/Jacq.http://en.wikipedia.org/wiki/Casearia#cite_note-GRIN-2


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Caseariais aplantgenusin thefamily@alicaceae. he genus was included in

theBlacourtiaceaeunder the *ron"uist systemof angiospermclassification, and earlier in

the @amydaceae. >ecent researchOcitation neededPindicates that the latter group might be reinstated as a

valid family.

hey are sometimes employed as honey plants, notably C. decandraandC. sylvestris. he latter

species is occasionally used as food by the caterpillarsof thewo:barred Blasher(*straptes

)ulgerator). @everal species are becoming rare due to deforestation. @ome appear close

toe!tinction, and C. +uinduensisof *olombiaandC. tini)oliafrom 6auritiusseem to be e!tinct

since some time in the &'th century and about %+H-, respectively.

lants traditionally used in India to treat fever or malaria were e!amined in vitro for

antiplasmodial properties against lasmodium falciparum. f ' analysed ethanol e!tracts, from

$H species, significant effects were found for 3% of the e!tracts. hese represent &3 different

species from &' families. f the active species &' were tested against . falciparum for the first

time. he following five species seems to be of special interest for further antimalarial studies,

*asearia elliptica, #olarrhena pubescens, ongamia pinnata, @oymida febrifuga, and lumbago

5eylanica.

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http://en.wikipedia.org/wiki/Planthttp://en.wikipedia.org/wiki/Genushttp://en.wikipedia.org/wiki/Family_(biology)http://en.wikipedia.org/wiki/Family_(biology)http://en.wikipedia.org/wiki/Salicaceaehttp://en.wikipedia.org/wiki/Flacourtiaceaehttp://en.wikipedia.org/wiki/Cronquist_systemhttp://en.wikipedia.org/wiki/Flowering_planthttp://en.wikipedia.org/wiki/Samydaceaehttp://en.wikipedia.org/wiki/Wikipedia:Citation_neededhttp://en.wikipedia.org/wiki/Wikipedia:Citation_neededhttp://en.wikipedia.org/wiki/Wikipedia:Citation_neededhttp://en.wikipedia.org/wiki/Honey_planthttp://en.wikipedia.org/w/index.php?title=Casearia_decandra&action=edit&redlink=1http://en.wikipedia.org/w/index.php?title=Casearia_sylvestris&action=edit&redlink=1http://en.wikipedia.org/w/index.php?title=Casearia_sylvestris&action=edit&redlink=1http://en.wikipedia.org/wiki/Caterpillarhttp://en.wikipedia.org/wiki/Caterpillarhttp://en.wikipedia.org/wiki/Two-barred_Flasherhttp://en.wikipedia.org/wiki/Deforestationhttp://en.wikipedia.org/wiki/Extincthttp://en.wikipedia.org/wiki/Extincthttp://en.wikipedia.org/wiki/Casearia_quinduensishttp://en.wikipedia.org/wiki/Colombiahttp://en.wikipedia.org/wiki/Casearia_tinifoliahttp://en.wikipedia.org/wiki/Casearia_tinifoliahttp://en.wikipedia.org/wiki/Mauritiushttp://en.wikipedia.org/wiki/Planthttp://en.wikipedia.org/wiki/Genushttp://en.wikipedia.org/wiki/Family_(biology)http://en.wikipedia.org/wiki/Salicaceaehttp://en.wikipedia.org/wiki/Flacourtiaceaehttp://en.wikipedia.org/wiki/Cronquist_systemhttp://en.wikipedia.org/wiki/Flowering_planthttp://en.wikipedia.org/wiki/Samydaceaehttp://en.wikipedia.org/wiki/Wikipedia:Citation_neededhttp://en.wikipedia.org/wiki/Honey_planthttp://en.wikipedia.org/w/index.php?title=Casearia_decandra&action=edit&redlink=1http://en.wikipedia.org/w/index.php?title=Casearia_sylvestris&action=edit&redlink=1http://en.wikipedia.org/wiki/Caterpillarhttp://en.wikipedia.org/wiki/Two-barred_Flasherhttp://en.wikipedia.org/wiki/Deforestationhttp://en.wikipedia.org/wiki/Extincthttp://en.wikipedia.org/wiki/Casearia_quinduensishttp://en.wikipedia.org/wiki/Colombiahttp://en.wikipedia.org/wiki/Casearia_tinifoliahttp://en.wikipedia.org/wiki/Mauritius


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-'A# $. *$RK:

he plan of work is divided into following steps8

Step /:

*ollection and identification of plant

Step 0:

reparation of whole plant e!tract.

Step 1:

@election of suitable e!tract for screening of pharmacological activities

Step 2:

?valuation of plant e!tract

%.#epatoprotective activity

*cl$ induced hepatoto!icity

aracetamol induced hepatoto!icity

hioacetamide induced hepatoto!icity

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+AT(R"A' A#% +(TH$%S

+aterials:

Selection of plant material

casearia elliptica leaf

(xperimental animals

All e!periments and protocols proposed in present study will be carried out according to the

guidelines of **@?A, =ew 1elhi and approved by the Institutional Animal ?thical *ommittee

(IA?*) .Albino Eistar rats of either se! (&'':&'gm) will be housed in group of - animals per

cage and maintained under standard laboratory conditions ( %&:h light;dark cycle, &$hrs). he

food in the form of dry pellets and water will be provided ad libitum.

Chemicals an! !rugs

*cl$,paracetamol,thioacetamide,indomethacin,caraggenan,diclofenac and all other chemicals to

be used in this project are of highly analytical grade.

S$RC( $. %ATA:

he source of data will be obtained from,

Laboratory based studies.

Literature survey, *1 >6.

=ational Q International 9ournals.

e!t books.

Internet.

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+etho! 0:-aracetamol in!uce! hepatotoxicity

#n the paraceta%ol ind!ced li$er to"icity %odel, paraceta%ol &1g()g i'p* dil!ted +ith

s!crose sol!tion +ill e ad%inistered daily for 7 days to all ani%als e"cept gro!p 1'

roup:%8 normal control.

roup:&8 standard

roup:38 #epatoto!ic rats with paracetamolRsucrose solution

roup:$8 >ats having hepatoto!icity treating with e!tract ofcasearia elliptica lea) Low dose.

roup:8 >ats having hepatoto!icity treating with e!tract ofcasearia elliptica lea) High dose.

-he ani%als +ill e sacri.ced 48hr after last treat%ent !nder light anesthetic ether'

/lood fro% each rat +ill e +ithdra+n y retro orital ple"!" !nder ether

anesthesia for ioche%ical in$estigation i' e' s-, sP-, s- and ilir!in

esti%ation' /lood +ill e allo+ed to coag!late at 370 and 30 %in and the ser!%

+ill e separated y centrif!gation at 2500 rp% for 15 %in' -he li$er of one ani%al

fro% each gro!p +ill e re%o$ed and processed i%%ediately for histopathological

in$estigation'

+etho! 1:Thioacetami!e in!uce! hepatotoxicity#n the thioaceta%ide ind!ced li$er to"icity %odel, thioaceta%ide &100%g()g s'c*

dil!ted +ith s!crose sol!tion +ill e ad%inistered daily for 7 days to all ani%alse"cept gro!p 1'

roup:%8 normal control.

roup:&8 standard

roup:38 #epatoto!ic rats with paracetamolRsucrose solution

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roup:$8 >ats having hepatoto!icity treating with e!tract ofcasearia elliptica lea) Low dose.

roup:8 >ats having hepatoto!icity treating with e!tract ofcasearia elliptica lea) High dose.

wenty four hours after administration of drugs, the animals were anaestheti5ed with anesthetic

ether for withdrawing the blood sample by cardiac puncture after over night of fast for

biochemical investigation i. e. s, s, sAL and bilirubin estimation. 2lood will be allowed to

coagulate at 3H'* and 3' min and the serum will be separated by centrifugation at &'' rpm for % min.

he liver of one animal from each group will be removed and processed immediately for

histopathological investigation.

Statistical analysis

he @tatistical analysis will be performed by using ne Eay A=ubia

cordifolia Linn. 9. ?thnopharmacol. &''-G %'38 $$/$+'.

&. *hatterjee 7. 6edicinal plants with hepatoprotective properties. In8 #erbal ptions. 3rd ?dn.

2ooks and Allied () Ltd. *alcutta. &'''G %3.

3. 7iritikar 7> and 2asu 21. Indian 6edicinal lants. International book distributors.1ehradun,

India. %++-. &'-$ / &'-.

$. 1ulaly *howdhury et al. Antimicrobial activity and cytoto!icity of Aerva lanata Bitoterapia.

&''&G H3G +&:+$.

. Vdupihille 6 and 9iffry 66. 1iuretic effect of Aerva lanata with water, normal saline and

coriander as controls. Indian 9 hysiol and harmacol. %+-G 3'8 +%:+H.-. >ao @ et al. ?valuation of an e!perimental model for studying urolithiasis effect of Aerva

lanata on urinary stones. Indian 1rugs. %+G &&8 -$':-$3.

H. 6erskey #, 2ogduk =. *lassifications of chronic pain8 1escription of chronic pain syndromes

and definition of pain terms. >eport by the International Association for the @tudy of ain ask

Borce on a!onomy. In8 6erskey #, 2ogduk =, editors. @eattle8 IA@ ressG %++$8&'+:%$.

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. 2ennett =, 2rown 69. =ervous @ystem8 ain and analgesics. In8 *linical harmacology.

%'th ed. ?dinburgh, @cotland8 *hurchill LivingstoneG &''8&+3:-.

+. 7umar obbins

and *otran athologic 2asis of 1isease, edited by 7umar ole of histamine in

rodent antinociception. 2r 9 harmacol %++$G%%%8%&-+:H+.

%$. 6almberg:Aiello , Lamberti *, Ipponi A, 2artolini A, @chunack E. ?vidence for

hypernociception induction following histamine #% receptor activation in rodents. Life @ci

%++G-38$-3:H-.

%. Lipnik:@tangelj 6, *arman:7r5an 6. #istamine:stimulated nerve growth factor secretion

from cultured astrocyctes is blocked by protein kinase * inhibitors. Inflamm >es &''$G3 @uppl

%8@H:.

%-. @tempelj 6, Berjan I. @ignaling pathway in nerve growth factor induced histamine release

from rat mast cells. Inflamm >es &''G$83$$:+.

%H.7okate *7, urohit A, okhale @2. &''&. e!tbook of pharmacognosy, =irali prakasan8

une. %8%:$ p. andurangan A, 7hosa >L, #emalatha @. &''. ?valuation of Anti:inflammatory

and Analgesic Activity of >oot ?!tract of Solanum rilo-atum Linn. Iranian 9ournal of

harmaceutical >esearch H(3)8&%H:&&%.

%.andurangan A, 7hosa >L, #emalatha @. &''+. ?valuation of anti:inflammatory activity of

the leaf e!tracts of solanum trilobatum Linn. 9 harm @ci >es %(%)8%-:&%.

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%+.arveen D, 1eng C, @aeed 67, 1ai >, Ahamad E, Cu C#. &''H. Antiinflammatory and

analgesic activities of hesium chinense urc5 e!tract and its major flavonoids, kaempferol and

kaempferol:3:o:glucoside. Cakugaku Dasshi %&H()8%&H:%&H+.

&'. Akdis *A, @imons B?>. #istamine receptors are hot in immunopharmacology. ur '

/harmacol. &''-G338-+ /H-.

&%. Akdis *A, 9utel 6, Akdis 6. >egulatory effects of histamine and histamine receptor

e!pression in human allergic immune responses. Chem $mmunol *llergy. &''G+$8-H/ &.

&&. @chneider ?, >olli:1erkinderen 6, Arock 6, 1y 6. rends in histamine research8 new

functions during immune responses and hematopoiesis. rends $mmunol. &''&G&38&/&-3.

&3. 2aena:*agnani *?. 1esloratadine activity in concurrent seasonal allergic rhinitis and

asthma.*llergy. &''%G-8&%/&H.

&$. Duberbier , Asero >, 2indslev:9ensen *. ?AA*I;A&L?=;?1B;EA guideline8

definition, classification and diagnosis of urticaria.*llergy.&''+G-$8%$% /%$&-.

&. hurmond >L, elfand ?E, 1unford 9. he role of histamine #% and #$ receptors in

allergic inflammation8 the search for new antihistamines.%at Rev rug iscov. &''GH8$%/3.

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s.s.j hepatoprotective protocol.docx