MINISTRY OF SCIENCE AND TECHNOLOGY
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E-mail: ured@znanost.hr
Title: Immunological disorders of eyes
Title: Molecular Oncology
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Title: Growth factor and Control of Cell Cycle
Title: Cell Receptors
Title: Activation of Early Genes: Immediate Cell Response
Title: Control of Cell Cycle: Cyclins and cdc Molecules
Title: Suppressor Genes
Title: Life and Death of Cell
Title: Cell Activation, Proliferation and Differentiation
Title: Kadherins as Receptors of Cell Adhesion
Title: Viruses in Control of Cell Proliferation
Title: Citostatics as Modulators of Tumor Cell Proliferation
Title: Genetical basis of genesis and development of cancer
Title: Regeneration
Title: Proliferation and Maturation of Embrional Cell at the
Different Stages of Mammalian Development
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Title: Vitamin C - Structure, chemical reactivity and biological
activity
Title: What is Molecular Medicine?
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Title: Expression and structure of gene for B chain of mouse
platelet derived growth factor
Faculty: Medicinski fakultet u Zagrebu
Author: SPAVENTI RADAN
Date of defense: 10/08/92
Language: hrvatski
Number of pages: 127
Summary: Using the RNA PCR we amplified 652 bp of 726 bp-long
codingsequence together with 684 bp of 3' noncoding sequence of mousePDGF B
cDNA. The cause of failure to amplify the whole codingsequence is located
in 30 and 44 base pairs long pieces which areparts of second and third
exon. In addition we presentedevidences that PDGF B from the normal mouse
tissue is notpolycistronic like it was suggested previously and
thatalternative splicing of sixth exon, known for PDGF A, in PDGF Bdoes not
occur. Analysis of PDGF B expression in mouse showedthat this gene is
activated in placenta, brain and heart tissue.The PDGF B expression in
Balb/c 3T3 cells which also posses thefunctional specific membrane
receptors suggest autocrine and/orparacrine growth control and presence of
same receptors onembryonic F9 cells, together with the fact that placenta
cellsproduce PDGF B, indicate the importance of this growth factorduring
embryonic development.
Keywords: PDGF B chain, RNA PCR
Title: Activity of c-myc gene in B chronic lymphatic leukemia.
Interrelation of proliferation and apoptosis
Faculty: Medicinski fakultet u Zagrebu
Author: POLJAK LJILJANA
Date of defense: 01/17/94
Language: hrvatski
Number of pages: 106
Summary: In spite of increasing body of evidence the mechanism
leading tothe accumulation of CD5+ B cells in the course of
chroniclymphocytic leukemia is still unresolved. Namely, it is
unclearwhether this accumulation results from increased cellproliferation
rate or delayed phisiologically programmed celldeath. Also, little is known
about the possible role of c-mycgene in these processes. In this study we
examined the activityof c-myc gene in peripheral B CLL cells in terms of
theirproliferation potential and the ability to activate apoptoticprocess.
We found that in most CLL cases the constitutionalactivity of c-myc gene in
CLL cells and normal cells did notdiffer. In addition, no correlation was
found among the degree ofc-myc gene activity and DNA synthesis rate in
leukemic cellscultured either in medium alone or in the presence of DMSO,
PMA,IL-2, PMA and IL-2, anti mi antibody or ionomicine. However thepositive
correlation was observed among c-myc gene activity andthe ability of
leukemic cells to activate apoptotic death whenthey have been cultured in
medium alone. Also, in the same cellsit was possible induce to the greater
extent apoptotic deathduring their in vitro cultivation by
metilprednisolone,ionomycine, PMA and anti mi antibody. On the other hand
cellsshoving the pattern of spontanous apoptotic death during in
vitrocultivation also showed the heterogeneity in the distribution
ofhybridization signal with bcl-2 DNA probe. These date suggest
theexistence of intraclonal heterogeneity between CLL cellssuggesting their
different potential for activation of apoptoticprocess. Also it is likely
that the delayed physiological deathof CLL could be responsible for their
accumulation rather thattheir increased proliferation rate likely could be
responsible.
Keywords: c-myc, chronic lymphocytic leukemia, apoptosis
Title: Immunohystochemical localization of oncoproteins in
colorectal tumors
Faculty: Medicinski fakultet u Zagrebu
Author: KAPITANOVIĆ SANJA
Date of defense: 09/21/92
Language: hrvatski
Number of pages: 60
Summary: There is increasing evidence suggesting that oncogenes have
animportant role in the pathogenesis of many malignant diseases.However
exact mode of their action in induction and maintenanceof transformation is
still unclear. In this study we investigatedthe expression of CEA and
erbB-2 oncoproteins in colorectalcarcinomas and adenomas using
immunohystochemical method. Alladenocarcinoma and adenoma specimens tested
were positive for CEAand erbB-2, but the intensity of staining was stronger
inadenocarcinoma tissue sections (++ or +++). Adenomas showedmoderate (++)
staining in the areas with the "atypical" cells,polypoide hyperplasia and
dysplasia, that means they could beprecancerous. Mucinous adenocarcinomas,
however, showed weak ormoderate immunoreactivity with both D-14 and
anti-erbB-2monoclonal antibodies. There was no correlation between
intensityof CEA oncoprotein expression and clinical stage (Dukes) of
thedisease, relapse and postoperative survival. However, thiscorrelation
was observed for erbB-2 oncoprotein. Its expressionwas strong or moderate
in all patients with Dukes C and D.Patients with Dukes A and B which showed
moderate or strongstaining had an earlier relapse of disease. There was one
patientwith adenocarcinoma Dukes B whose lymph node was negative
inconventional (hematoxylin and eosin) testing. However, when thesame lymph
node was tested by use of immunohystochemical methodtumor cells, positive
to erbB-2 oncoprotein were found.Therefore, it seems that expression of
erbB-2 oncoprotein couldbe the prognostic factor in rectosygmoid
adenocarcinomas.
Keywords: colorectal tumors, erbB-2, CEA
Title: Characterization of substance immunologicaly
cross-reactive with insulin isolated from murine melanoma cells B16BL6
Faculty: Sveučilište u Zagrebu, smjer Biologija u Zagrebu
Author: ŠARIĆ TOMO
Date of defense: 06/25/93
Language: hrvatski
Number of pages: 110
Summary: The association of hypoglycemia with tumors other than
insulinomais one of known endocrine syndromes associated with neoplasia.The
most obvious mechanism by which a tumor can producehypoglycemia is by
secretion of insulin or some insulin-likesubstance. Also, the high
molecular weight Substance(s) differentfrom, but Immunologically
Cross-Reactive with Insulin (SICRI) hasbeen proposed as a cause for
tumor-hypoglycemia. In a variety oftumor-bearing patients and animals SICRI
was detected by means ofinsulin-specific radioimmunoassay (RIA). Murine
melanoma B16BL6was shown to be a rich source of SICRI and because of this
it waschosen as in vitro model for SICRI-studies. Attempts to purifySICRI
to homogenity from melanoma cells or to establish a methodfor
SICRI-detection failed to give specific and consistentresults. Because of
these discrepances, the specificity of thepositive insulin-RIA measurements
in murine melanoma B16BL6extract was examined. It was shown by
trichloroacetic acidprecipitation assay, thin layer
cellulose-chromatography andelectrophoresis, as well as by FPLC, that
SICRI-activity inmelanoma cells is a positive artifact due to radiolabeled
insulindegradation in RIA. Positive RIA-readings have been
erroneouslyascribed to SICRI. What was really measured was not a
newsubstance immunologicaly cross-reactive with insulin, but loss
ofimmunoreactivity of insulin tracer due to its proteolyticdegradation. The
eexperiments with protease inhibitors show thatthe enzyme most probably
responsible for insulin-degradation isinsulin-degrading enzyme.These
findingsdeny the existanse ofSICRI in melanoma B16BL6 cells.In the future,
appropriateexperiments with enzyme inhibitors could be performed to
checkother possible tumor sources for SICRI-activity. Always,validation of
the RIA should include convincung proof that traceris not degraded and thet
other possible nonspecific effects areruled out.
Keywords: melanoma, insulin, insulin degrading enzyme
Title: Collagenase type IV in tumor invasion and metastasis
Faculty: Sveučilište u Zagrebu, smjer Biologija u Zagrebu
Author: DESPOT-SLADE NEDA
Date of defense: 05/17/93
Language: hrvatski
Number of pages: 74
Summary: The collagenase-like enzymes with the ability to degrade
theproteins of the artificial basement membranes (BM) was isolatedfrom
human invasive fibrosarcoma. The secretion of the samepeptide was observed
from the primary established fibrosarcomacell culture. This peptide
degrades the artificial basementmembranes derived from bovine corneal
endothelial cells. Usingthe electrophoretical methods it was found taht the
isolated andpartially purified enzyme consists of 8 bands of
differentmolecular masses corresponding to the collagenase standard fromCl.
histolyticum. Between them, only two bands with molecularmasses of 22000
and 63000 degrade basement membrane. Exposure ofinvasive cell line to TGF
beta abrogates destruction of BM. TGFbeta inhibits collagenase activity and
stimulates specificmetalloproteinase inhibitor (TIMP) in invasive tumor
cells. Wepreasume that TGF beta could play a protective role in
tumorinvasion.Production of type IV collagenase by tumor cells hasbeen
linked to their metastatic potential in several experimentalmodels. This
enzyme is responsible for basement membranedestruction. In order to find
out the importance of our findings,the affinity purified monoclonal
antibodies against human 72000type IV collagenase were employed by
immunohistochemicaltechnique. In planocellular and baseocellular skin
carcinomas wedidn't find any correlation between expression of this enzyme
andmetastatic potential. Probably it is because of high invasive,but not
matastatic, potential of baseocellular carcinomas.However, in endometrial
carcinomas there is direct correlationbetween the intensivity of
immunohistochemical staining for typeIV collagenase and the depth of
myometrial invasion. The extentof myometrial invasion is significant
indicator of metastaticpotential. High percentage of positive cells in
invasivecarcinomas support the role of the enzyme in the process
ofmetastasis.To find the possible correlation between type IVcollagenase
expression and TGF beta, the in situ hybridizationwith TGF beta 1 cDNA was
performed on the same specimens thatwere already tested on type IV
collagenase. The intensivity ofhybridization signal for TGF beta 1 was
compared with intensivityof immunoreaction on type IV collagenase. In
specimens that werenegative or weakly positive on type IV collagenase, the
signalobtained by hybridization with TGF beta 1 cDNA was stronger thanin
control cells. The cDNA is linked for TGF beta 1 gene and onlyone copy
gives the signal, the intensive staining could beexplained as gene
amplification.The augmented number of TGF beta1 gene copies could give
higher expression in vivo. The higherexpression of TGF beta could stimulate
TIMP and inhibits type IVcollagenase activity. It is in agreement with our
findings invitro. Gene amplification (higher expression of protein) could
beresponsible for lower metastatic potential of some tumorspecimens.
Keywords: Type IV collagenase, metastasis, tumor, TGF beta 1
Title: Automated oligonucleotides synthesis and their
purification
Faculty: Sveučilište u Zagrebu, smjer Biologija u Zagrebu
Date of defense: 12/09/92
Language: hrvatski
Number of pages: 80
Summary: Purification of different-lenght-oligonucleotides
byreversed-phase HPLC and polyacrylamide gel electrophoresis wascompared
with the aim of determining the advantages anddisadvantages of these two
purification methods. Criteria used toevaluate the successfulness of
purification were: recoveredoligonucleotide yield, DNA purity,
time-consumption and costrationality. The results showed that
reversed-phase HPLCpurification is a more appropriate method for
oligonucleotidepurification: it is fast, gives high resolution of sample
and thesample yield is in average 4.5 times higher than
afterelectrophoretic purification. Purification of oligonucleotides bygel
electrophoresis did not require expensive apparatus and gavepure and
homogeneous products but it required a large quantity ofthe sample and is
much more time consuming.
Title: Immunohistochemical detection of c-myc oncoprotein in
breast tumors.
Faculty: Prirodoslovno-matematički Zagreb
Number of pages: 65
Keywords: c-myc, breast cancer, immunohistochemistry
Title: Expression and structure fe gene for B chain of mouse
platelet derived growth factor
Faculty: Medicinski fakultet u Zagrebu
Date of defense: 10/08/92
Number of pages: 127
Author: Spaventi Radan
Degree level: Ph.D.
Title: Activity of c-myc gene in B chronic lymphatic leukemia.
Interrelation of proliferation and apoptosis.
Faculty: Medicinski fakultet u Zagrebu
Date of defense: 01/17/94
Number of pages: 106
Author: Poljak Ljiljana
Degree level: Ph.D.
Title:
Number of pages: 0
Author: Grazio Simeon
Degree level: M.A.
Title:
Number of pages: 0
Author: Kapitanović Sanja
Degree level: M.A.
Title:
Number of pages: 0
Author: Pećina-Šlaus Nives
Degree level: M.A.
Title:
Number of pages: 0
Author: Slade Neda
Degree level: M.A.
Title:
Number of pages: 0
Author: Šarić Tomo
Degree level: M.A.
Title:
Faculty: Prirodoslovno-matematički Zagreb
Mentor: PAVELIĆ KREŠIMIR
Date of defense: 07/06/94
Number of pages: 60
Author: Horvatić Marijeta
Degree level: M.A.
Title:
Faculty: Prirodoslovno-matematički Zagreb
Mentor: PAVELIĆ KREŠIMIR
Author: Matijević mr. Vesna
Degree level: M.A.
Title:
Faculty: Prirodoslovno-matematički Zagreb
Mentor: PAVELIĆ KREŠIMIR
Date of defense: 01/20/95
Number of pages: 57
Author: Gall-Trošelj Koraljka
Degree level: M.A.
Title:
Faculty: Prirodoslovno-matematički Zagreb
Mentor: PAVELIĆ KREŠIMIR
Date of defense: 01/23/95
Number of pages: 61
Author: Hraščan mr. Reno
Degree level: M.A.
Title:
Faculty: Prirodoslovno-matematički Zagreb
Mentor: PAVELIĆ KREŠIMIR
Date of defense: 01/31/95
Number of pages: 50
Author: Pečur mr. Lada
Degree level: M.A.
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