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Related post: lyophilization to concentrate the factor were employed. This experimental
design was followed throughout to establish the characteristic logarithmic
and stationary periods of growth and to time the occurrence of the factor.
The sedimented cells resulting from these centrifugations were washed
3 times, sonicated, resuspended, and tested for lytic effect. Other
bacterial species, e.g.. Bacillus subtilis , Escherichia coli , and P^.
aeruginosa were also examined. The density of Buy Bimatoprost Ophthalmic Solution 0.03 the cultures was determined
with a Klett-Summerson colorimeter and they were grown on agar plates to
determine the colony count. Dialysis tubing of different porosities were
used. High performance liquid chromatography (HPLC) and Sephadex gel
filtration were employed in preliminary purification trials. The flagel-
lated Buy Bimatoprost Ophthalmic Solution species, Leishmania brasiliensis , Trypanosoma rhodesiense , T^.
equiperdum , and 2 other strains of T_. cruzi were examined for their
susceptibility to the ATF, and other bacterial substances (lipopoly-
saccharides from Salmonella typhosa , S^. typhymurium , E^. coll , and
Serratia marcescens were assessed for lytic effects comparable to those
of P^. f luorescens . In vivo studies were performed employing mice inoculated
intraperitoneally with the Tulahuen strain of T^. cruzi and intramuscular
injections of Bimatoprost Cost the ATF suspended in phosphate buffer. Blood smears, tissue
imprints, and histologically processed tissues were stained with Giemsa.
Our findings disclosed the extracellular nature of the ATF; this was
clearly indicated by its production during the logarithmic period of
growth and its absence from washed, sonicated cells. Of the bacterial
species studied, only P^. aeruginosa produced lytic effects comparable
to those of P^. f luorescens , and endotoxins Bimatoprost Buy from the bacterial species
mentioned above did not produce lysis of the trypanosomes. All the
flagellated species studied were susceptible to the P^. f luorescens ATF.
HPLC and Sephadex gel filtration disclosed the occurrence of at least
one active fraction, but the exact molecular weight and chemical
characterization are currently being assessed. Preliminary estimates
obtained with dialysis tubing filtration indicated a molecule between
6,000 and 12,000 daltons. Infected mice treated with 2 or 3 doses of
ATF (25 to 200 yg protein) exhibited a rapid drop in the parasite count
(within 24 hours) and liver and spleen of the treated animals did not
exhibit the considerable necrosis and parasitic foci observed in the
nontreated, parasitized controls. A marked release of white cells was
25-27
Serial No. ZOI AI 00097-22 LPD
observed both in infected and uninfected controls) which was always
reproducible; this release subsided with time.
The immediate proposed course of this work is to purify the ATF
and to repeat the in vivo experiments using the purified fraction. However,
a more extensive elucidation of questions related to the cause of the
drop in parasitemia in the treated animals, the marked leucocyte release,
and the mechanism of action of the ATF would enhance our knowledge and
understanding not Bimatoprost Ophthalmic Solution 0.03 Buy only of the ATF but also of the enzymic function of
the parasite membrane which is an important aspect of this project.
The observed relationship between a trypanosome and a bacterium
provides another interesting example of microbial interaction. In
view of the wide public health significance of Chagas ' disease, these
findings are also of potential importance in parasite chemotherapy
and may aid further in our understanding of the cellular biology and
physiology of flagellar motility.
Publications ;
Mercado, T. I. and Garbus, J. 1979. Creatine phosphokinase isoenzymes
and Trypanosoma cruzi infections. Comp. Biochem. Physiol . 6AB : 11-15.
25-28
SMITHSONIAN SCIENCE INFORMATION EXCHANGE
PROJECT NUMBER (Oo NOT use this space)
U.S. DEPARTMENT OF
HEALTH, EDUCATION, AND WELFARE
PUBLIC HEALTH SERVICE
NOTICE OF
INTRAMURAL RESEARCH PROJECT
PROJECT NUMBER
ZOl AI 00098-24 LPD
PERIOD COVERED
October 1, 1979 to September 30, 1980
TITLE OF PROJECT (80 characters or less)
Biochemical mechanisms of energy metabolism in mammalian and parasitic
organisms .
NAMES, LABORATORY AND INSTITUTE AFFILIATIONS, AND TITLES OF PRINCIPAL INVESTIGATORS AND ALL OTHER
PROFESSIONAL PERSONNEL ENGAGED ON THE PROJECT
PI: E.G. Weinbach Head, Physiology and Biochemistry Section, LPD, NIAID
Other: L.S. Diamond Head, Parasite Growth and Differentiation Section, LPD,
NIAID
C.E. Glaggett Biological Laboratory Technician (Biochem.), LPD, NIAID
D.B. Keister Biologist, LPD, NIAID
D.M. Dwyer Research Microbiologist, LPD, NIAID
COOPERATING UNITS (if any)
Laboratory of Chemical Physics, NIAMDD (H. Kon) ; Metabolism Branch, NGI
(D. Tschudy and P. Ebert) .
lab/branch
Laboratory of Parasitic Diseases
SECTION
Physiology and Biochemistry
INSTITUTE AND LOCATION
NIAID, Bethesda. Maryland 20205
TOTAL MANYEARS:
36/12
PROFESSIONAL:
22/12
1 4/1?
CHECK APPROPRIATE BOX(ES)
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