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GEORGI
M E DICAI 4

NEWS

No 5 (218) 2013

Published in cooperation with and under the patronage

of the Thilisi State Medical University

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GEORGIAN MEDICAL NEWS

No 5 (118)1011

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1J 060ux C/f"y'(aJIX wmro 8iJJpaxeH xapaxmepHblu d!lR II.O.cDTJ( nux npu J. 286 HoM 3. Barba
Mer lanll
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4. MacK,
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 GEORGIAN MfD1CAL Nt;W S
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DJlaronapHOCTb. HacToIIW3..11 paGora BblnOl1HeHa npl-! support for wound healing. Alternative Medicine Review
¢lHHaHcoBo~ nOAJlep)f(Ke HaUHOHaJ1bHoro HaY'tHOro 2003; 8(4): 3 59-377.
QloHlla rpY3HH HM. W. PYCTaOel1H B pa~H;ax rpaHr a AR· 5. Rode D. Com frey tox ic ity revi sited. Trends Phannacol.
109·8·403-11. So:. 2002 ; 1.3(11): 497-499.

JlHTEPATYPA SUMMARY

1. BarbakadzeY., KernertelidzeE., Targamadzel., Mulkijan­ DEVELOPMENT OF TECHNOLOGY FOR THE

yanK., ShashkovA.S., UsovA. J. Poly[3-(3,4-dihydroxy­
phenyl)glyceric acid], a new biologically active polymer
from Symphylum asperum Lepech. and Scaucas;cum Bieb.
(Boraginaceae). Molecules 2005; 10{9): 1135-1144.
2. Barbakadze V., Mulkijanyan K., Gogilashvili L., Amira­
nashvili L., Merlani M., Novikova Zh., Sulakvelidzc M..
Allantoin- and pyrrol izidine alkaloids-free: wound healing
3 20
compositions from Symphytum asperum. Bull. Georg. Nal).
u/J. Acad. Sci. 2009; 3( 1): 159-164.
3. Barbakadze v., Gog i l as h~ i l i L., Am i t a n ash~ili L.,
Merlani M., Mulkijanyan K. Spc:ctrophotomer1ric quan­
HQMCTP titat ive detennination ofpoly(3-(3 ,4-dihydroxyp henyl)
lj') :l00­ glyceric acid), Bull. Georg. Natl . Acad. Sci . 2010; 4(3):
123·1 26.
4. MacKay D., Miller A. L. Wound healing. Nutritional
O CM'"
SUBSTANCE OF POLYI3 - (3,4.DIHYDROXYPHF.­
NYL) GLYCERIC ACID I FROM SYMPHYTUM
ASPERUM
Gokadze S., Barbakadze V., Goglluhvili L.,Amiranash­
vill L., nakuridze A.
TbifiJi Siale Medical University. Tbilisi, Georgia
Comfrey lSymphyrum L.) is used to treat bone fractures,
tendon injuries. ulcer lesions of gasnoimeslinal tract. It
promotes wound healing, accelerates exudates resorption
in lungs and reduces joints' inflammation . In Georgian
fo lk medicine, herbal remedies from comfrey are used to
accelerate regeneration processes. Comfrey contains hepa­
7S

totoxic and carcinogenic pyrrolizidine alkaloids. besides the
mai n active ingredient is poly [3 - (3,4-d ihydroxypheny l)
glyceric acid] (pDPGA).
The aim of present work was to develop a technology for
the substance -poly (3-(3,4dihydroxyphenyl) glyceric acid]
(PDPGA) from comfrey stems, free of tox ic pyrrolizidine
alkaloids.
During the investigation the optimal conditions for extrac­
tion and purification have been established: on the first stage
pyrrolizidine alkaloids were removed from plan t material
by supercritical extraction; then the crude polysaccharides '
fraction was obtaioed by water extraction (raw materials!
cxtragent ratio was I: 15 at 900c, the procedure was carricd
twice for 60 and 90 minutes). The isolation of the final
product - PDPGA from crude polysaccharides' fraction was
carried out by ultrn.fllrration on membrane 11 lters. Based on
the results of the investigation the tcchnological scheme
for the substance has been developed.
Keywords: com frey stems, substance poly (3-(3,4-dihy­
droxyphenyl) glyceric acid) (pDPGA) extraction.
P E310ME
PAlPAfiOTICA TEXHOJIonm: nOJlY'lEHIDl C Yli­
('' TAHl.\IUI nOJl" 13-(3.4-)(HrH.lO'OKC"<I>EHHJI)
rJlHl.\EPH HOBoA KHCJ10Tbll H3 SYMPHYTUM
A$PERUM
r OI<'a.ue C.H., Ga p6aKlUlle B.B., r ornn aUl Blin H JI.M .,
AMllpaHaWBHJIII A.W., fi BK)'pHD.le A ..Il.*.
T6W1ucocuu ~opcm 6el(HblU .wet}U'IUHI.:KUU y"u6epcu­
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renaroTOk'CK'lecKOro If KaplUtHOI"CH.Horo lleAcnUHI. ..po­
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(II.1l¢n<).
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76
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b. ~a:p::of'lb ),j,1.~! bfl.).-,=,xn
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yc,10alul JJtCTpaku)tH Ii O\( Hcn:H q6CTaHuiIll n ,n<l>rK:
HB nepeoM )Tane 3KCTpakUHIf H3 pacrnTeJfloHOro CblpWl
allKanO llAOB rpy nnl.l nHppOlllllllll,Jl Ha npoH3BO..llUTC1l
C)I(H"lkeHKWMH ralaMli. larCh! HlBnC\(eHHe CYMMbI norutca­
xap"JtOtl, II COOTHoweHHH Cl.I.phII H)kcrpareHTOII I: I 5 npH
TeMnep arype 900c, ABa)l(AbI - II Te\(eHHe 60 H 90 MllltyT.
BblJteJleHlle XOIlC'IHOro npollY1CT8 - n.ll.<l>rK H3 CYMMbI
nO.'JKcaxapHllo a npoHlBOAKTCIf ynloTPa$KJtI.Tpaulien Ha
h!cM6pallKblx cptl.JTbTPax. n Olly<teH llloIe pelynl>T'ilTbl HC­
cnCliOBaHH$I lIel'Slit B OCHOOY TexHOnOrH\(eCKOA CxeMlJ
nonY'IeKHlI cy6cnmwllt.
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'Oa6 0~)e,~oua~060u 3"PJ'::'u] u{Jo il6.::.6(joou ao~..,
0011 6ad6" )~ ' 'l(')oo l (JaaVll~3ao~
b. e,M.i.~Oa. a · t\.::.~~d')oJ,~. llM(')o~ ,::,'330~r.,
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m~o~(lllOu h,)hJ ~aV(l'OM u.::.a~(lOn6M l)ooaa(l',·
u06a6(1, (I)~os.r:.'ob<'l, u.::..i.::.fJ(J133~M
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~f',3J~oO 2000-'bO aa6n Vs.!!:'o u e,.::.6H'::'3~f'l~')'00
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 GEORGIAN MEDICAL NEWS
No S (2/8) ]OJJ
6'Jc41.:.'bO' M~~a(1)3W"'W - 60 ~ ... 90 V'O'H,tJ ~.:.6a.r
a~MO",'aO. J(<)~ou"'J,,~oWlloOU )!.-.8oJ,:S,l.::.6 J~~a- ou
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OtIb ~,)a«ltla6a6oO').
STUDY OF EFFECTIVENESS OF BIOLUMINESCENT REPORTER PHAGE ASSAY
ON Y. PSEUDOTUBERCULOSIS STRAINS
Mitaj~hvili N.
1. JCNokhishYili Tbtlisi State University, Tbilisl Georgia
Yersinla i:J a genus ofbacteria in the family Enterobacteri·
aceae, Yersiniae are Gram-negative facultative anaerobic
bacilli. Yersinia pseudotuberculosis causes Far East
scarlet~like
fever in hwnans, wbo occasionally get infected
zoonotically, most often through the food-borne route
(II]. The designation pseudoruberculosis derives from the
characteristic histopathological aspect found in mesenteric
lymph nodes oflymphoid hyperplasia cases which closely
resemble those observed during tuberculosis infection. Hu­
mans develop varying degrees of illness, from abdominal
pain and fever to septicemia. but a mesenteric adcnitis that
mimics an acute appendicular syndrome (pseudoappendi­
I.. citis) is the most common clinical presentAtion, as well as
gastroenteritis [2,9, I 0]. Other complications include lesions
" to lymph nodes, spleen and liver. The disease is most com­
,­
moo in children and young adults (6,8) and immunocom­
I. promised individuals are at greater risk ofsevere disease or
death. Isolation and identification of Y. pseudotuberculruis
""
is difficult and time consuming because of presence other
gut associated bacteria. Thus, there is an urgent need to
r.
develop pseudotuberculosis rapid detection techniques.
'" Reporter phage assay is rapid detection technique and its
efficiency is not affected by presence of contaminating
bacteria and no sample preparation is needed.
Reporter phage assay: Recombinant reponer phage may
provide a "nalural" and specific approach for the detection
of r.pseudotuberculosis. Recombinant "light-tagged"
(U ' reporter phage was genetically engineered by integrating
the Vibrio haney; fuxAB genes into the genome of Y. pestis
,..... .
specific phage. The phage is mixed with bacterial cells, the pseudotuberculosis strains NR-4J71, NR-4372, NR-4373, .
phage infects the cell, iuxAB are expressed, and the cell
bioluminesces when substrate decanal is added. Sample
processing is not necessary; the phage and cells are mixed,
incubated at appropriate temperature and subsequently
luminescence is measured using luminometer. Reporter
phage-mediated detection systems have been developed for
Lysreria monocylogenes, salmonella, mycobacteria. Bacil­
1'" lus anthracis, Escherichia coli 0 I 57;H7 and Yersinia pestis.
(.' . ,. The CDC plague-diagnostic phage cJ)A I 122 was used as a
COM'"
J31!!."J3ob 71D~OMoO b.,)'B'lJd3~,')~ W'::'DS!?M uvo·
u61>60oou aO~DOob 6Dd6M~M~O'lJ~ u,:fua"u.
tool fortbe identification of r pestis. [7]. Previous studies
have demonstrated that the wild-type cJ)A 1122 phage can
lyse nearly all natural r pestis isolates, and is 'specific'
for Y. pestis (1.3,4 }; however, some Y. pseudOlubercu/osis
strains have been shown to be cJ)AII22 susceptible when
grown at temperatures above 20°C [1 ,3,5). The reasoD
for the temperature-sensitive differential susceptibility is
unknown, but presumably due to temperature-dependent
changes in the cell surface layers/composition. Therefore,
a potential caveat of the reporter phage detection system
is the possibility of a false-positive response with strains
from the closely-related species of Y. pseudotuberculosIs.
Performing the repon er phage assay at the restrictive
temperature (20°C) will prevent a false positive signal
in samples that may contain Y. pseudotuberculosis. Thus,
specificity can be strictly controlled when using isolated
cultures grown at a specific temperature (12}. The presented
study was conducted at the Department of Molecular Ge­
netics and Microbiology, Emerging Pathogens Institute,
College of Medicine, Univcrsity of Florida, USA, within
the framework ofUNESCO/Japan Young Researcher' Fel­
lowships Programme, 2012.
The aim of this study was to study the effectiveness of
bioluminescent reporter $A 1122 phage assay on strains of
Y. pseudotuberculosis and other different Yersinia specie~
in presence of other enteric bacteria and to detennine the
optimal condit ions (time, temperature, bacterial number).
Material and methods. Bacterial strains andphage. The Y.
NR-4l74, NR-4l75, NR-4380, NR-804, .nd NR-2l207
were obtained from the Biodefense and Emerging Infec­
tions Research Resources Repository. Y.pseudOlubercu/osis
CR·OOOI from Emerging Pathogens Institute Building,
University of Florida; Nine strains representing different
Yersinia species: Y. rucker; ATCC 29473, Y. frederiksenll
ATCC 33641. Y. kristensen;i ATCC 33638, .Y. rohdei
ATCC 43380. Y. bercovier; ATCC 43970, Y. enterocolitica
ATCC 9610, Y. aldovae ATCC 35236. Y. mollarelii ATCC
77