PROGRAM

We have a great pleasure of inviting you and your colleagues to this symposium 
Prof. dr. Mladen Belicza and Prof. dr. Križan Čuljak


Organizacijski odbor Organizing Committee

President: Prof. Dr. Križan Čuljak 
Department of General Pathology and Pathological Morphology of the Veterinary Faculty,  Zagreb, Heinzelova 55, Croatia 
Tel: 385-01-2390-310, Fax: 385-01-214-697
Vice-president: Prof. Dr. Mladen Belicza  
"Ljudevit ]urak" Clinical Department of Pathology University Hospital "Sestre milosrdnice",  Zagreb, Vinogradska 29 Croatia 
Tel/fax: 385-01-572-892
Secretary: Prof. dr. Ruža Sabočanec, tel. 385-01-2390-311 
Dr. sci. Božo Krušlin, tel/fax: 385-01-572-892 or 187-244
Scientific Board
Prof. dt E Silvestri (Italy) Prof. dr. Ž. Dolinar (Switzerland)
Prof. dr. M. Štulhofer (Croatia) Prof. dr. W Walkiewicz (Poland)
Prof. dr. K. Lapis (Hungary) Prof. dr. H. G. Fassbender (Germany)
Prof. dr. M. Bielicza (Croatia) Prof. dr. K. Čuljak (Croatia)
Prof. dr. A. Pirkić (Croatia) Prof. dr. D. Ferluga (Slovenia)
Prof. dr. Z. Biđin (Croatia) Prof. dr. L. Šenk (Slovenia)
Prof. dr. Z. Kusić (Croatia) Prof. dr. Jasna Talan-Hranilović (Croatia)
Location of the Symposium 
MUZEJ MIMARA, ZAGREB, F. ROOSEVELTA 5

  
 
RASPORED PREDAVANJA     PROGRAM
Petak (Friday), 06. 06. 1997.
9,00  Otvaranje skupa Opening ceremony
 Memorijalna predavanja Memorial lectures
9,15 Bussani R, Pavletić N, Mannone T, Silvestri F (Trieste) 
Unusual coronary lesions causing sudden or unexpected death.
9,30 Coffee break Odmor
A. Pathological morphology of human and animal diseases 
     Patološka morfologija humanih i animalnih bolesti 
  Chairmen - Predsjedatelji: Silvestri F, D. Ferluga, K. Čuljak
10,00 Planinc D, Sutlić Ž, Biočina B, Huseđinović I, Anić D, Belicza M, Ferenčić Ž, Čupić H, Jelić l, Sokolić J (Zagreb): Heart transplantation - the Zagreb experience. 
10,10 Čupić H, Belicza M, Ferenčić Ž, Planinc D, Sutlić Ž, Biočina B, Anić D (Znagreb) Endomyocardinl biopsy in cardiac graft pathology. 
10,20 Bussani R, Mannone T, Pavletić N, Silvestri F (Trieste) Tumours metastatic to the heart and pericardium.
10.30 Vučić M, Zaninović K, Cvjetko I, Žurić M, Dumenčić B, Periz R, Plasaj T (Zagreb)Utility of "Thanatos" computer system in the evaluation of causes of death regarding meteorologicnl variations. 
10.40 Discussion and coffee break   Rasprava i odmor 
 Chairmen - Predsjedatelji: Pirkić A, Grabarević Ž, Šoštarić B
11.00 Krušlin B, Pirkić A, Matković M, Klarić P (Zagreb) Inununohistochernistry of borderline ovarian tumours (LMP).
11.10 Nola M, Jukić S, Ilić-Forko J, Babić D, Kos M, Užarević B, Petrovečki M (Zagreb)Values of horrnone steroid receptors, hormonal factors and DNA content in patients with endometrial carcinoma before and after treatment with tamoxifen. 
 11.20 Jandrić D, Belicza M, Kurjak A, Šumski R, Latin V (Zagreb) Comparisou of the pathological and clinical findings of placentas in Department of Gynaecology and Obstetrics of the General Hospital "Sveti Duh" in Zagreb. 
11.30 Glumbić I, Lež C, Pirkić A (Zagreb) Chorangioma placentae.
11.40 Juntes P (Ljubljana)  Evaluation of morphometrical data on AgNOR area and number in the simple and complex type adenocarcinomas of the mammary gland in the dog.
11.50 Lipej Z, Šoštarić B (Zagreb) Pathomorphological findings in two cases of swine Iymphosarcoma.
12.00 Oraić D, Zrnčić S, Soštarić B (Zagreb)  Enteric Redmouth disease in rainbow trouth (Orecorlcynchtcs mykiss)
12.10 Heinzel R, Banek LJ, Belicza M (Zagreb) Comparison of' numeric and volume density of apoptotic bodies with - frequency of apoptotic bodies in 10 HPF in prostatic hyperplasia.
12.20 Eminović-Behrem S, Jonjić N (Rijeka) DNA ploidy pattern and nucleolar organiser regions (AgNORs) as prognostic factors in colorectal cancers.
12.30 Tomasović Č, Habek D, Pirkić A (Zagreb)  Endocervical carcinoma metastatic to a "symplastic" myoma.
12.40 Reljić A, Krušlin B, Trnski D, Jandrić D, Čupić H, Belicza M (Zagreb)  Carcinosarcoma of the urinary bladder: A case report.
12.50 Torlaković E, Litz CE, Mc Clure, Brunning RC (Minnesota) Tri-color fish/immunophenotyping study of Yh-chromosome in B-lymhlrocytes.
13.00 Zrnčić S, Oraić D, Šoštarić B (Zagreb) "Winter disease" in cage cultured sea bream (Sparus aurata).
13.10 Tomasović Č, Čupić H, Petric V, Ivkić M, Krušlin B (Zagreb) Basaloid squamous cell carcinoma of the larynx: A case report.
13.20 Manojlović S, Levanat S, Pavelić B, Crnić I (Zagreb)Morphological and genetic changes in odontogenic cysts. 
13.30 Discussion and coffee break   Rasprava i odmor 
Chairmen - Presjedatelji: Talan-Hranilović J, Sabočanec R, Jonjić N
15.00  Talan-Hranilović J, Sajko T, Kogler A, Lambaša S, Kalousek M (Zagreb) Ki G7 - antigen in choroid plexus papillomas and carcinomas.
15.10 Mrčela M, Dmitrović B, Hlavka V (Osijek, Zagreb) Heterotopia of tlre cerebral grey matter. A case report.
15.20 Torlaković E, Cherwitz D, Scholes J, Jessurn H, Mc Glennen R (Minnesota) A comlarison of B-cell gene rearrangements in gastric lymphoma and chronic gastritis - A polimerase chain reaction study.
15.30 Jonjić N, Kovač D, Krašević M (Rijeka) Prognostic signiticance of microvessel density in acivanced gastric cancer.
15.40 Gumzej Ž, Meštrić A, Brezovečki D, Belicza M (Zagreb)  HIP gastritis: We did not find any correlation between grades of activity and intensity of mucosa colonisation by HP.
15.50 Torlaković E, Snover DC (Minnesota)  "Serrated adenomatosis coli" A new type of adenomatous polyposis of the colon.
16.00 Krušlin B, L.ukač J, Kovačević D, Kordić D, Marković Lj, Tomasović Č, Kusić Z, Belicza M (Zagreb)  C-erb B-2 oncoprotein in patients with colorectal carcinomas.
16.10 Discussion and coffee break   Rasprava i odmor 
B. Iatrogenic, experimental and environmental pathology  
Iatrogena, eksperimentalna i patologija okoliša

Chairmen - Presjedatelji: FerenčićŽ, Šenk L, Ćurić S
16.30 Sabolić I, Herak-Kramberger CM, Brown D (Zagreb, Boston)  Cadmium induces morphological and functional changes of cells in the rat epididymis.
16.40 Herak-Kramberger CM, Sabolić I, Thevenod F (Zagreb, Homburg-Saar)  Selective effects of cadmium upon plasma membrane proteins in the rat proxunal tubule.
16.50 Černe M, Zorman Rojs O (Ljubljana)  Comparison of pathohistologic changes on chicken organs experimentally inoculated with highly virulent field isolate and vaccinal strain of infectious bursal disease virus (IBDV).
17.00 Modrić Z, Grabarević Ž, Milas Z, Bratulić M (Zagreb) Pathomorphological changes in acute attack and latent phase of experimental eperithrozoonosys of swine.
17.10 Tišljar M, Grabarević Ž, Artuković B, Bratulić M, Džaja P, Sikirić P, Seiwerth S (Zagreb)  The influence of BPC 157 on nitric oxide autagonist and agonist induced lesions in broiler chicks.
17.20 Božić Z (Zagreb)  Scientifically established approach to the research of endemic uropathy.
17.30 Discussion and pause   Rasprava i odmor 
19.30 Group dinner    Zajednička večera 
Subota (Saturday) 07. 06. 1997.
C. "Herman Jurak" round table on rheumathological diseases 
"Herman Jurak"- okrugli stol o reumatološkim bolestima

Chairmen - Presjedatelji: Fassbender HG, Djorđević G, Vizjak A 
9.10 Fassbender HG (Mainz) Pathology of cardiac processes in rheumatic fever. 
9.30 Vizjak A, Ferluga D, Perković T, Rozman B (Ljubljana)  Skeletal muscle immune deposit in systemic lahus erythematosus: correlation with histologic changes and autoantibodies.
9.40 Djorđević G, Krstulja M, Jonjić N (Rijeka) Microvessel quantitation in synovium. 
9.50 Discussion and coffee break   Rasprava i odmor 
D. Ethical forum - Forensic medicine: Profession? Science? Politics? 
     Etički forum - Sudska medicina: Struka? Znanost? PoIitika?

Chairmen - Predsjedatelji: Andelinović Š, Marcikić M, Jantes P 
10,10 Dugački V (Zagreb) Lifetime and work of Prof. dr Ljudevit Jurak. 
10,20 Kaić Z, Brkić H, Mandušić J (Zagreb)  Forensic dentistry (past, present, future). 
10,30 Marcikić M, Mandić N (Osijek)  Unusual mode of disposal of dead body. 
10.40 Anđelinović Š, Primorac D (Split)  Molecular biology techniques in forensic medicine
10.50 Šoštarić B, Lipej Z, Stanin D, Gomerčić N (Zagreb) The tragedy of one bear (Ursus arctos) from Lika.
11.00 Šipuš N (Zagreb)  Causes of death of Roman Emperors.
11.10 Discussion and coffee break   Rasprava i odmor 
E. Computer systems aud computerisation in pathology  
     Informatički sustavi i kompjuterizacija u patologiji

Chairmen - Predsjedatelji: Plasaj T, Belicza M, Brezovečki D
11.30 Vučić M, Žurić M, Brezovečki D, Periz R, Plasaj T (Zagreb) Comparison of clinical and pathological diagnosis by "Thanatos" computer system.
11.40 Žurić M, Vučić M, Čupić H, Krušlin B, Belicza M, Periz R, Plasaj T, Zurak K, lvkić M, Kusić Z (Zagreb) Capabilities of "Thanatos" computer system in creating of specific register for thyroid diseases.
11.50 Vučić M, Žurić M, Brezovečki D, Belicza M, Periz R, Plasaj T, Kusić Z (Zagreb) "Thanatos"pathological data pool as a part of the hospital cancer register. 
12.00 Žurić M, Šimunović D, Čupić H, Plasaj T, Kusić Z (Zagreb) Distribution of thyroid carcinomas according to age, sex and pathological diagnosis - comparison of a two five-year periods.
12.10 Discussion and coffee break   Rasprava i odmor 
F. Seminars in diagnostic immunohistonathology 
     Metode u patologiji
12.30 Bumber-Bolanča S (Zagreb) LSAB visual system (DAKO).
12.44 Završetak skupa Closing ceremony

 

BOOK OF ABSTRACTS 

EDITORS 
Božo Krušlin M.D., Ph.D. 
Mirna Robić M. Sc


ACKNOWLEDGMENTS 
Excellent technical assistance of Dr. Mirna Žurić, resident of pathological anatomy and Miss. Renata Periz, secretary of "Ljudevit Jurak" Department of Clinical Pathology, "Sestre milosrdnice" University Hospital Zagreb, in preparing of this book of abstracts should be cordially appreciated 
Editors

UNUSUAL CORONARY LESIONS CAUSING SUDDEN OR UNEXPECTED DEATH

Bussani R, Pavletić N, Mannone T, Silvestri F

lnstitute of Pathological Anatomy - University of Trieste - Italy

We describe some unusual autopsy findings in four patients with sudden or unexpected death. 
CASE N.1: A 46-year old man was admitted to Hospital because of a suspected aseptic necrosis of the femur. In the course of hospitalisation the patient evidenced signs of severe pulmonary hypertension and died. At autopsy a massive, bilateral pulmonary thromboembolism was found. The thrombotic material reached the right atrium, passed through the left ventricle to lodge in the circumflex left coronary artery. CASE N.2: An 89-year old woman was hospitalised for ischemic cardiopathy and anaemia. During gastroscopy the patient suddenly collapsed and died. At autopsy a gigant Lambl's excrescence of the closure line of the free edge of the right coronary ostium was evident. The heart presented a large myocardial infarction involving the posterior wall of the left ventricle and the posterior third of the interventricular septum. 
CASE N.3: A 28-year old man was found dead at home by family members. At autopsy the heart showed no significant atherosclerosis, nevertheless a large myocardial infarction of the anterior wall of the left ventric(e and the anterior third of the interventricular septum was found. Microscopically, a lymphocytic coronaritis of the left descending artery was present. The inflammatory process involved neither other coronary branches nor other vascular districts. There was no histological evidence of myocarditis. 
CASE N.4: A 39 year old man sufćering of Prinzmetal's angina died suddenly. At autopsy, histologic sections of the left descendenting artery of the left coronary artery showed diffuse eosinophilic adventitial granulomatous inflammation. Large areas of active lymphocytic myocarditis were biventricularly evident.
A. Pathological morphology of human and animal diseases

    Patološka morfologija humanih i animalnih bolesti
HEART TRANSPLANTATION - ZAGREB EXPERIENCE

Planinc D, Sutlić Ž, Biočina B, Huseđinonić I, Anić D, Belicza M, Ferenčić Ž, Čupić H, Jelić I, Sokolić J

University Hospltal "Sestre milosrdnice", Zagreb, Croatia University Hospital "Dubrava", Zagreb, Croatia
University Hospital "Rebro", Zagreb, Croatia

From 30. September 1988. to 30. April 1997, 36 patients (29 men and 7 women), aged from 19 to 62, underwent orthotopic heart transplantation (Htx). lndications for heart transplantation's were as follows: in 27 patients dilatative cardiomyopathy, in 8 patients ishaemic cardiomyopathy and in one patient ishaemic cardiomyopathy with severe aortal and mitral valve failure. Ejection fraction of left ventricle before transplantation was 8 to 24 % (mean value 16.2%), and PRV 1.3 to 6.5 Wood' s units (mean value 2.8). ' 
All patients were receiving immunosuppressive drugs like antithymocytic globulin (ATG), cyclosporine A, azathioprine and steroids. Diagnosis of grade of cardiac rejection was based on histological findings of right ventricle endomyocardium (based on ME Billingham criteria and 1SHT). Immunosuppression was successfully achieved in 40 moderate cardiac rejection processes, with high doses of metilprednisolone and ATC. Ctinically manifest infections were established in 16 cases (4 of them lethal) and complications and adverse effects of immunosuppressive therapy in 74 cases (3.4 per patient). 
After Htx patients were fotlowed on the average for 29 months. Nine patients died in early and nine in late postoperative period, 15 patients had arterial hypertension (mostly cnild to moderate) and two had renal insufiiciency of grade 1-Il. The function of graft was satisfactory or excellent in 18 surviving patients (three of them are working full- time) and the quality of life is very good. Ejection fraction of left ventricle after Htx is 35 to 68% (average 55%) and patients are in NYHA class I-II. 
 

ENDOMYOCARDIAL BIOPSY IN CARDIAC GRAFT PATHOLOGY

Čupić H, Belicza M, Ferenčić Ž, Planine D, Sutlić Ž, Biočina B, Anić D

KB "Sestre Milosrdnice, KB "Dubrava ", KBC "Rebro" , ZAGREB

Thirty six orthotopic heart transplant procedures were perfonned in the nine years between September 1988 and April 1997 . In order to determine the degree of cardiac rejection, hematoxylin-eosin and Mallory-stained sections of endomyocardial biopsies (EMB) were analysed. Specimens were graded according to the International Society for Heart Transplantation classification (since April 1991) , which is essentially an extension of the original M.E . Billingham criteria. 
To make an adequate sample, four pieces of myocardium are required, 50% of which must be evaluable myocardium and not a previous biopsy site or scar. Additional information is given for inadequacy of specimens (less than four 
pieces), "Quilty" effect, contraction bands and perimyocitic fibrosis. 
In spite of intensive search for simple non-invasive rejection diagnostic procedures with a view to replacing or reducing the number of EMB, EMB of transplanted heart remains important for establishing the grade of graft rejection in management of the posttransplant patients. 
 

TUMOURS METATATIC TO THE HEART AND PERICARDIUM

Bussani R, Mannone T, Pavletic N, Silvestri F

Institute of Pathological Anatonry - University of Trieste - Italy.

Tumour metastases to the heart are one to the least investigated subjects in clinical oncology. Nevertheless, this problem should retain great practical significance being connected with operability as well as with the type of treatment of neoplastic patients. 
Of 4,796 patients autopsied at the Institute of Pathological Anatomy of Trieste from 1994 through 1995, 1,148 males (42%) and 780 (38%) had a primary tumour. Among them 162 subjects had cardiac metastases (8,4% of all cancers), with a significant frequency decrease in the elderly patients. Mesothelioma, melanoma and lung cancers showed the highest frequency of heart involvement in males (100%, 50%, and 31% respectively). Melanoma, lung tumours and renal neoplasm's had the highest percentages of heart secondary lesions in females (45%, 26%, and 20% respectively). 
Among lung cancer, anaplastic small cell carcinoma (37%), and adenocarcinoma (33%) showed the most frequent heart involvement in males, while squamous carcinoma (43%) and anaplastic small cell carcinoma (29%) had the highest frequency of heart metastases in females. 
Adenocarcinoma (82%) and anaplastic small cell carcinoma (74%) of the lung, mesothelioma (100%) and breast tumours (73%) had the highest frequency of metastases to the pericardium. 
A particularly high frequency of myocardial involvement was observed in squamous cell carcinoma of the lung (62%), urinary tract tumours (60%) and melanoma (45%).

 

UTILITY OF "THANATOS" COMPUTER SYSTEM IN THE EVALUATION OF CAUSES OF DEATH REGARDING METEREOLOGICAL VARIATIONS

Vučić M, Zaninović K, Cvjetko 1, Žurić M, Dumenčić B, Periz R, Plasaj T

"Ljudevit Jurak" Clinical Department of Pathology, University Hospital "Sestre milosrdnice", Zagreb

Using archives of "Ljudevit Jurak" Clinical Department of Pathology we have reviewed data on died patients in the University Hospital "Sestre milosrdnice" in 1995 and 1996. The main cause of death and main pathological condition were determined for each case, according to recommendations of the World Health Organisation published in the International Classification of Diseases and Related Health Problems (tenth revision, 1994). All data were memorised in, and analysed by "Thanatos" computer system for clinical pathology. 
During mentioned period 2566 patients died in hospital, 1269 in 1995 (average age 67), and 1297 in 1996 (average age 69). Diseases of cardiac and vascular system were the most common cause of death, where only myocardial infarct (MI) has caused 1 17 deaths (4,5% of total). 
MI is considered to have many provoking risk factors, among which are also listed weather changes. This has stimulated us to correlate, by using "Thanatos" computer system, frequency of deaths caused by MI with three weather parameters average air temperature, average air pressure and average relative humidity. 
Correlation of mentioned pathological and weather data was done on a basis of comparison of seven-day periods throughout both years. Graphical analysis has clearly revealed three peaks of Ml deaths, followed by two distinctive patterns of weather changes. In the period 28 Jan. - 3 Feb. 1995 average air temperature, and average air pressure were increased while average relative humidity was lower in comparison with periods before and after MI death peak. On the other hand, in the periods 8-14 Apr. 1995, and 23-29 Nov. 1996, average air temperature was lower, while average air pressure and average relative humidity did not differ in relation to periods before and after MI death peaks. 
Although our population was not random(y selected (hospital patients), presented results indicate necessity of further investigation of influence of weather changes on human athophysiology.

IMMUNOHISTOCHEMISTRY OF BORDERLINE OVARIAN TUMOURS

Krušlin B, Pirkić A, Matković M, Klarić P, Hodek B

Clinical Department of Pathology "Ljudevit Jurak" and Department of Gynecology and Obstretics, University Hospital "Sestre milosrdnice ", Zagreb, Crontia

In the period 1990-95 one hundred forty eight malignant epithelia! ovarian neoplasm's were diagnosed, ten of whom were borderline tumours (7 serous and 3 mucinous). Borderline epithelial tumours of the ovary (tumours of low malignant potential, LMP) still represent a great problem due to the lack of uniform pathohistological criteria and uncertain biological behaviour. Additional criteria for evaluation and prediction of biological behaviour of these tumours are obviously needed. In this study, we investigate proliferative kinetics of cells of LMP tumours. It is well known that these tumours, particularly mucinous, very often contain interspersed areas of well-differentiated as well as atypical cells. 
Expression of proliferative antigens (Ki-67 and PCNA) was studied using immunohistochemical methods Strept AB/AP and Strept AB/HRP after microwave antigen retrieval treatment. We also investigated the expression of c-erbB-2 and p53 by the same methods. 
In this study 8 LMP tumours were analysed. Two patients were lost from follow-up and were not included. Positive nuclear staining for Ki-67 was observed in 5 specimens. This reaction showed mosaic pattern, and was strongly positive in the areas showing epithelial atypia. PCNA nuclear positivity was found in 6 cases, independently of the degree of cell differentiation. Positive staining with c-erbB-2 protein antibody was obtained on the cell membrane in 5 tumours, and was mostly expressed in the areas showing eytological atypia. Positive nuclear reaction to p53 oncoprotein with mosaic pattern was found in 4 cases, particularly in the areas showing atypical epithelium. 
The expression of investigated antibodies in all specimens was strongly positive in areas showing cytological atypia and dedifferentiation, except of PCNA that showed positivity also in the areas without proliferative activity. Different expression of proliferative antigens was probably due to their regulation of the various phases of cell cycle. 
Expression of p53 and c-erbB-2 in investigated tumours probably indicate their potential aggressiveness. 
Our results of immunohistochemical expression of investigated antigens may be useful as an additional criteria in morphological evaluation of LMP ovarian neoplasm's, and may have a prognostic significance. Further studies of larger series are certainly needed.

REFERENCES

  1. Kiyokawa T. Int J Gynecol Pathol 1994;13:311
  2. Wang DP et al. Virchows Arch [A] 1992;421:393
  3. Hall PA et al. J Pathol 1990;162:285

VALUES OF HORMONE STEROID RECEPTORS, HORMONAL FACTORS AND DNA CONTENT IN PATIENTS WITH ENDOMETRIAL CARCINOMA BEFORE AND AFTER TREATMENT WITH TAMOXIFEN

Nola M, Jukić S, Ilić-Forko J, Babić D, Kos M, Užarević B, Petronečki M

Department of  Gynaecological and Perinatal Pathology, Clinical Hospital Centre, Zagreb, Croatia

Introduction. Tarnoxifen is a non-steroidal triphenylethylene derivate with predominant anti-oestrogen activity. 
Aims. To determine the influence of tamoxifen on hormone steroid receptors, hormonal status and values of DNA content in patients operated for endometrial carcinoma. Thirty patients with endometrial carcinoma were treated with tamoxifen in dose of 30 mg for 10 days after curettage. 
Methods. During diagnostic curettage hormonal status (luteinizing hormone (LH), follicle-stimulating hormone (FSH), progesterone, estradiol, testosterone, androstendione, dehydroepiandrosterone sulphate (DHEAS), sex hormone binding globulin (SHBG)), hormone steroid receptors (oestrogen and progesterone receptors) and DNA content (DNA ploidy, proliferative activity and DNA-index) were estimated. On the day of operation levels of the same parameters were determined again. 
Results and Conclusions. The level of PH were significantly higher and the level of ER were significantly lower after the tamoxifen treatment. This was detected in the group of patients with deep myometrial invasion, clinical stage 1, high and moderate degree of differentiation and favourable histologic type. The statistical difference was not detected according to DNA content. Hormonal therapy consisting of combination of progestine and tamoxifen can be improved if we focus our investigation on finding out parameters of prognostic significance.

 

COMPARISON OF PATHOLOGICAL AND CLINICAL FINDINGS OF PLACENTAS AT DEPARTMENT OF GYNAECOLOGY AND OBSTETRICS OF GENERAL HOSPITAL "SVETI DUH" IN ZAGREB.

Jandrić D, Belicza M, Kurjak A, Šumski R, Latin V

Departmerrt of Gynaecology and Obstetric of General Hospital "Sveti Duh", Zagreb

From routine biopsy material from maternity ward of General Hospital "Sveti Duh" in Zagreb a number of placentas have been extracted and correlation of pathological and clinical findings from pathological pregnancies and deliveries has been done. The intention of the scientific work was to present possibilities of researching placental pathology in routine diagnostic material. Preliminary research shows that most common clinical conditions due to which placentas were sent to pathological analysis are next: 
a) Acute hypoxia of fetus with which compensatory angiomatosis of chorionic villi was found, more abundant and emphasised sincicial knots, in fewer number of cases infarct of placenta and in one case large retroplacental hematoma. 
b) With placentas delivered by caesarean section was noticed compensatory angiomatosis of chorionic villi and emphasised sincicial knots . 
c) With hypotrophia of fetus main pathological substrate was branching disorder of chorionic villi with lack of branches of I and II rank (Hiatus villorum), then more abundant perivillosus fibrinoid and calcifications. In several placentas compensatory angiomatosis of chorionic villi was found. 
d) Several placentas connected with trisomy 18 and 21 were processed. Disordered maturation with chorionic villi of immature type was found by histology , as well as branching disorder of chorionic villi (Hiatus villorum), in several placentas only one umbilical artery was found, and in almost all placentas, infiltration of decidua capsularis was found. 
e) In analysis of premature delivery completely developed chorioamnionitis was found.

CHORANGIOMA PLACENTAE

Glumbić I, Lež C, Pirkić A

"Ljudevit Jurak" Clinical Departnaent of Pathology, University Hospital "Sestre milosrdnice", Zagreb

Chorangioma placentae was considered as a very rare tumour of placenta, but in the recent literature its frequency is about 1% (1). Being classified as an haemangioma, its histological appearance is variable and pleomorphic. It could be divided into endotheliomatous, capillarious, cavernous and fibromatous form, from which the capillarious is the most common of all chorangiomas. Chorangiomata probably arise as malformations of the primitive angioblastic tissue of the early palcenta. There are many reported instances of risk pregnancies, which were accompanied with the placental chorangiomata, such as hydramnion, pre-eclampsia, antepartum bleeding, premature onset of labour, but the overall perinatal mortality is only minimal increased. On the contrary, clinically significant chorangiomas, greater then 4 cm, are associated with fetal loss rate of approximately 40%. 
Recently, we have observed chorangioma as an incidental finding during the routine macroscopic examination of the placenta of the 28 year primipara whose pregnancy was clinically normal. She was admitted to the hospital for labour at 40 weeks of gestation. Pregnancy, labor and delivery were uncomplicated. She was delivered of a healthy female child measuring 52 cm, 3500 g, with Apgar score 1=9, 5=10. Placenta measured 16:16:2,5 em, 350 g, with centrally attached umbilical cord and radially distributed alantoic blood vessels. Solitary, ovoid mass was observed on the fetal side of the placenta, measuring 4 cm in greatest diameter with soft and bright red cut surface. 
Histologically, our case of chorangioma was of the capillarious appearance, and in this manner, the immunohistoreactivity to UEA lectin was strongly expressed in tumours tissue. Despite to the clinical data, we believe that transient early pregnancy hypoxia have triggered a circumscribed excessive angioblastic proliferation in the placental tissue. 
 
  1. Wallenburg HCS. Chorangioma of the placenta: thirteen new cases and a review of the literature from 1939 to 1970 with special reference to clinical complications. Obstet and Gynecol Survey, 1971;26:411-425.
  2. Quintero R, Reich H, Johnson MP, Gonacalves L, Evans MI. In utero endoscopic devascularisation of a large chorangioma. Ultrasound Obstet Gynecol, 1996, 8/1:48-52.

EVALUATION OF MORPHOMETRICAL DATA ON AgNOR AREA AND NUMBER IN THE SIMPLE AND COMPLEX TYPE ADENOCARCINOMAS OF THE MAMMARY GLAND IN THE DOG

Juntes P

Institute of Pathology, Veterinary faculty Ljubljana, Slovenia

Silver staining of nucleolar organizer regions (NORs), or better their artyrophilic proteins (AgNORs), is a method used in tumour pathology for evaluation of the speed of tumour cell proliferation. The quantity of AgNor proteins is in corre(ation to the ratio of cells in cell cycle as evaiated with Ki-67 and PCNA in some studies. WHO classification discriminate two main groups of tumours of the mammary gland in the dog - simple and complex type. AgNOR area and number were morphometrically analyzed in adenocarcinoma of simple and complex type using semiquantitative image analysis system Prodit 5.2 (BMA, The Netherlands). Significant differences were found between both types of tumours, for total mean AgNOR area (P=0.0541). The mean number of AgNORs and maximal number of AgNORs were significantly higher in simple type adenocarcinomas (P=0.0000, P=0.0012), and were in correlation with total mean AgNOR area (r--0.4379, P=0.0002; r=0.3485, P=0.0004). Total mean AgNOR area in simple type tumours was in negative correlations with two years postsurgical period without tumour (r=-0.4002, P=0.0284) and so was the mean area of an average AgNOR granule (r=-0.4534, P=0.0119) and maximal area (r--0.4340, P=0.0166). The number of AgNORs was in correlation with the number of mitoses (r=0.3109, P=0.0111).

 

ENTERIC REDMOUTH DISEASE IN RAINBOW TROUTH (Onchorhyncus mykiss)

Oraić D, Zrnčić S, Šoštarić B

Croatian Veterinary Institute

Enteric redmouth (ERM), an acute to chronic septicemic disease affects all species of salmonids and is caused by the bacteria Yersinia ruckeri from the family Enterobacteraceae. Losses caused by this disease might reach up to 70% of overall losses in all salmonid fish farm and 25% of the fish can be infected but asymptomatic. The bacteria Yersinia ruckeri was isolated from diseased rainbow trout having clinical appearance of ERM at several fish farms in Croatia. The aim is to describe clinical behavior, pathological changes and bacteriological findings of endophtalmitis and encephalitis characterized by presence of bacterial organism compatible with Yersinia ruckeri. The skin of afFected fish was darkly pigmented, with unilateral or bilateral exophtalmia. They swam slowly, close to the water surface, and were separated from the rest of the lot. Some of affected fish presented obvious CNS signs, such as as slow convulsice swimming. Reaction to stimulation of any type (such as feeding or alarm reaction) were markedly reduced. On pathoanatomical examination marbled liver, transparent and hemorrhaged intestines, pronounced hemorrhage on swim bladder and enlarged spleen were most consistent findings.

COMPARISON OF NUMERIC AND VOLUME DENSITY OF APOPTOTIC BODIES WITH FREQUENCY OF APOPTOTIC BODIES IN 10 HPF IN PROSTATIC HYPERPLASIA

Heinzl R, Banek Lj, Belicza M

University Hospital Dubrava, Zagreb, Croatia

As a controlling mechanism of programmed cell death, apoptosis is important factor in tissue growth kinetics, not only in tumours but also in normal and hyperplastic altered tissues. This process can be stimulate or stopped by numerous endogenous and exogenous factors which are (most of them, probably) in direct connection with tissue proliferative activity, controlled by this way. Apoptosis is morphologically in light microscopy clearly visible as apoptotic cell or apoptotic body - round or oval acidophylic cell or body containing nuclear fragments as basophilic particles - so it is available to morphologic and quantitative analysis. 
Considering apoptosis as a process complementary to mitotic activity, it is clear that evaluation of "apoptotic index" should be of practical importance in estimation of tissue growth kinetics (especially in tumours and hyperplastic tissues) and in following up its malignant or premalignant potentia) - just as mitotic index already is. 
For daily routine practice, it is important for procedure of evaluating "apoptotic index" to be enough reliable and quick (and cheap) versus relatively complicated and slow stereologic methods (evaluating of numeric or volume densities, for example). lmage - analysis is still available for a few pathohistologic laboratories. This problem can be resolved very simply - by counting apoptotic bodies in 10 HPF - just as for mitotic index. Because these are complementary parameters, in this way they should be compared quite well. 
ln this work we compared results of counting apoptotic bodies in 10 HPF with results of evaluating numeric and volume densities in hyperplastic prostate - glandular, basal and atypical (P1N II) type of hyperplasia. Numeric and volume densities were highest in basal hyperplasia group, something lower but statistically significant in PIN 11 and the lowest in simple glandular group. 
Same results we obtained by counting apoptotic bodies in 10 HPF. Our results confirm reliability of this method of "apoptotic index" evaluating; so we suggest to employ this term and this method of its evaluating in routine practice.

LITERATURE:

  1. Kyprianou N, Tu H, Jacobs SC. Apoptotic versus proliferative activities in human benign prostatic hyperplasia. Hum Pathol 1996; 27(7);668-75.
  2. Baličević D. The significance of apoptotic bodies appearance in prostatic hyperplasia and carcinoma. (Magisterij) Zagreb: Medicinski fakultet Sveučilišta u Zagrebu, 1993. 108 str.
  3. Šarčević B, Belicza M, Šeparović V. Apoptosis in breast cancer. Libri Oncol 1992; 21(12)43-8.
  4. Heinzl R. Evaluation of apoptosis in hyperplastic prostata. (Magisterij, u izradi).

DNA PLOIDY PATTERN AND NUCLEOLAR ORGANISER REGIONS (AgNORs) AS PROGNOSTIC FACTORS IN COLORECTAL CARCINOMAS

Eminović-Behrem S, Jonjić N

Department of Pathology, Faculty of Medicine, University of Rijeka, Rijeka, Croatia

Ninety-four surgically removed colorectal carcinomas were processed for DNA flow cytometric analysis and one-step silver method was used for staining of interphase nucleolar organiser regions (NORs). The aim was to investigate the prognostic significance of DNA ploidy and AgNOR and the relationships to histological grade and Dukes' stage. In addition, the s-phase value was related to Dukes' stage, histologic grade, DNA ploidy and AgNOR count. There were no significant correiation's between ploidy pattern, histological grade and Dukes' stage. However, diploid tumours had significantly lower AgNOR counts (<5) than aneuploid tumours, 
Furthermore, Dukes' C stage tumours exhibited higher AgNOR counts than Dukes' A and E tumours. Any significant correlation between s-phase and the other observed parameters were found. lndividually, none of the parameters under study influenced survival rate except Dukes' stage. However, if tumours were first divided into groups with low AgNOR counts (<5) and groups with high AgNOR counts (<S), the correlation with survival was significant for histological grade Dukes' stage and ploidy pattern, in the group with high AgNOR count.

 

ENDOCERVICAL CARCINOMA METASTATIC TO A "SYMPLASTIC" MYOMA

Tomasović Č, Habek D, Bolf-Benković L, Pirkić A

"Ljudevit Jurak" Clinical Department of Pathology, University Hospital "Sestre Milosrdnice", Zagreb

Metastasis of cervical carcinoma in uterine leiomyoma is a rare event, and it was seldom reported about. Recently, we have observed one unusual case of invasive cervical carcinoma spreading into the endocervically sited leiomyoma, that was histologically confirmed as atypical (or simplastic) leiomyoma. We could not find any data in the available literature concerning this matter. 
We report a case of a 74 years old patient who underwent surgery due to cervical carcinoma, presented at stage Ib. Surgical specimen showed infiltrative tumour of the cervix, measuring I,Scm in greatest diameter and three solid, firm, white, sharply circumscribed nodules. Two of the nodules were intramural, up to 3 cm in greatest diameter, located in body of uterus and were histologically confirmed as leiomyomas with marked hyalinisation. The third nodule was located in the endocervix, close to the infiltrative tumour, measuring 1cm in greatest diameter. Histologically it was composed of bizarre tumour cells. Variation in size and shape, hypercromatic nuclei and multinucleated forms were present, but no increased mitotic activity. Within this tumour small solid nests of atypical epithelial cells resembling those in cervical carcinoma were found. 
An extensive immunohistochemical examination (pancytokeratin, cytokeratin 10/13, desmin, PCNA, Ki-67 and p53 antibodies) was provided, and the given results were discussed in the scope of different proliferation potential of both tumours.

CARCINOSARCOMA OF THE URINARY BLADER: A CASE REPORT

Reljić A, Krušlin B, Trnski D, Jandrić D, Čupić H, Belicza M.

Departments of Urology and Pathology, University Hospital "Sestre milosrdnice", Zagreb, Croatia

We report a case of carcinosarcoma of the urinary bladder in a 72 years old male patient. The patient presented with painless, intermittent hematuria lasting for three months. Clinical examination revealed a (arge, bulky, partially necrotic, tumours mass involving the whole cavity of urinary bladder. Pathologic analysis of transurethrally resected tissue revealed a tumour composed of both carcinomatous and sarcomatous component. The tumour was predominantly composed of moderately to poorly differentiated, atypical transitional epithelial cells forming nests and islands infiltrating submucosa and muscu(ar layer. Between these nests, areas of abnormal cartilage suggesting chondrosarcoma were observed. Immunohistochemical analysis showed positive reaction with anti-cytokeratin antibodies and epithelial membrane antigen in areas of epithelial differentiation and positivity for S-100 protein and vimentin in areas of atypical cartilage. The patient underwent total cystoprostatectomy. The urinary bladder was completely infiltrated with whitish, partially necrotic tumorous tissue, measuring 10 cm in largest diameter. Pathohistologic analysis was consistent with previous findings. In the bladder wall, a diverticulum measuring 3 cm in largest diameter filled with tumours tissue was observed. Histologic analysis revealed tumours tissue showing only epithelial difterentiation. The patient developed recurrence 6 months after the initial presentation and was treated by surgery. 
 

REFERENCES

  1. Lahoti C et al. Urology 1994;43:389
  2. Young RH. Cancer 1987;59:1333
  3. Holtz F et al. Cancer 1972;29:294
  4. Young RH, Eble JN. Hum Pathol 1992;22:948 5. Delides GS. Br J Urol 1972;44:571

TRICOLOUR FISH/IMMUNOPHENOTYPING STUDY OF Ph-CHROMOSOME IN B-LYMPHOCYTES

Torlaković E, Litz CE, Mc Clure J, Brunning RC

Department of Laboratory Medicine and Pathology, University of Minnesota Medical School, Minneapolis, Minnesota

Chronic myelogenous leukaemia (CML) is a myeloproliferative disorder characterised by a neutrophilic hyperleukocytosis, basophilia and the t(9;22), Philadelphia chromosome, in the myeloid cells. Although this leukaemia manifests as a myeloid proliferation, it may undergo transformation to a precursor B-cell acute lymphoblastic leukaemia. This has led to the hypothesis that the oncogenic event in CML occurs in a pluripotential hematopoetic stem cell capable of both myeloid and lymphoid differentiation. 
While myeloid differentiation of the stem cell has been directly demonstrated, evidence of lymphoid differentiation has been indirect. This has been due primarily to the low number of lymphocytes present in the marrow and blood of patients with CML. 
This study examines the Philadelphia chromosome status in CD20 and CD3 positive lymphocytes from CML patients employing a tricolour florescence immunophenotyping IFISH technique. Terminal deoxynucleotidyltransferase (Tdt) positive and myeloperoxidase positive cells were also examined for the presence of the Philadelphia chromosome. 
Six patients with previously diagnosed chronic myelogenous leukaemia were studied by a tricolour immunophenotyping/FISH method for direct determination of the Philadelphia chromosome in B- and T-lymphocytes. Two patients had involvement of CD 20 positive lymphocytes. CD3 positive lymphocytes in al) patients were negative for the Philadelphia chromosome.

 

BASALOID SQUAMOUS CELL CARCINOMA OF THE LARYNX: A CASE REPORT

Tomasović Č, Petric V, Čupić H, Ivkić M, Krušlin B

Clinical Departments of Pathology and Otorhinolaryngology, University Hospital "Sestre milosrdnice", Zagreb, Croatia

We report two patients with basaloid squamous cell carcinoma (BSCC) of the larynx. Both patients were males ageing 62 and 65 years, respectively. They were heavy smokers and moderate drinkers. They presented with hoarseness of three months' duration and dysphagia. Patients presented in Stage I and 1V, respectively. One patient was treated by surgery including radical dissection of the neck and the other by partial laryngectomy. The later patient underwent surgery 5 years earlier, due to colorectal carcinoma. At present, the patients are well, with no signs of the disease 3 and 6 months after diagnosis. BSCC is a distinctive, rare, highly aggressive variant of squamous cell carcinoma that arises in a variety of anatomic sites, including the larynx. So far, less than 30 cases of BSCC of the larynx were reported in the literature.The tumours were composed of moderately pleomorphic basaloid cells forming nests, cords, and cribriform patterns. Foci of necrosis, and squamous differentiation as well as small cystic spaces containing mucinlike material, were also observed. Surface mucous showed squamous dysplasia. BSCC should be distinguished from adenoid cystic carcinoma and small cell carcinoma usually in routine hematoxylin and eosin stained sections, or in rare cases with the aid of immunohistochemical methods. Their distinction is very important because adenoid cystic carcinoma is much less aggressive than BSCC. A careful follow-up of the patients is clearly needed. 
 

REFERENCES

  1.  Banks ER et al. Am J Surg Pathol 1992;16:939
  2.  Wain SL, Kier R, Vollmer RT, Bossen EH. Hum Pathol 1986;17:1158

Ki 67 - ANTIGEN IN CHOROID PLEXUS PAPILLOMAS AND CARCINOMAS

Talan - Hranilović J, Sajko T, Kogler A, Lambaša S, Kalousek M

"Ljudevit Jurak" Clinical Department of Pathology University Hospital "Sestre milosrdnice", Zagreb, Croatia

Reactivity of Ki 67-antigen was tested in six choroid plexus tumours (three papillomas and three carcinomas) in purpose to evaluate the proliferative potential of each tumour. All evaluated tumours were diagnosed in male children ageing from few months to 12 years except one papilloma which was diagnosed in 23 years old female. 
Retrospective immunohistochemical characterisation and evaluation of the proliferative potential in this three papillomas and three carcinomas of choroid plexus was performed by reaction on Ki67 - antigen using Strept ABC complex/HRP duet visualisation system method. DAKO Rabbit Anti - Human Ki 67-antigen on the paraffm sections was used. 
The purpose of this evaluation was to determine and to quantify the proliferative potential in each papilloma and also to compare the proliferative potential of papillomas in relation to carcinomas. Our results show the existence of significant differences in the number of positive immunoreactive cells within choroid plexus papillomas and especially choroid plexus carcinomas. 
Quantitative differences of Ki 67 - antigen nuclear reactivity may prove as useful clinical indicator for prognosis of the progression and recurrence of choroid plexus tumours. 
 

REFERENCES:

  1. Kato T, Fujita M, Jamura Y, Tada M, Abe H, Nagashima K, Nakamura N: Clinicopathological study of choroid plexus tumors: immunohistochemical features and evaluation of proliferative potential by PCNA and Ki-67. NOSHUYO - BYORI 1996; Nov 13 (2): 99 - 105
  2. Vajtai I, Varga Z, Aguzzi A: MIB-1 immunoreactivity reveals different labelling in low - grade and in malignant epithelial neoplasms of the choroid plexus. HISTOPATHOLOGY 1996; Aug 29 (2): 147 - 51
  3. Karamitopoulou E, Perentes E, Diamantis I, Maraziotis T: Ki-67 immunoreactivity in human central nervous system tumors: a study with MIB1 monoclonal antibody on archival material. ACTA NEUROPATHOL 1994; 87: 47 - 54
  4. Louis D N, Edgerton S, Thor A D, Hedley - Whyte E T: Proliferating cell nuclear antigen and Ki-67 immunohistochemistry in brain tumours: a comparative study. ACTA NEUROPATHOL 1991; 81: 675 - 679

A COMPARISON OF B-CELL GENE REARRANGEMENTS IN GASTRIC LYMPHOMA AND CHRONIC GASTRITIS - A POLYMERASE CHAIN REACTION STUDY

Torlakarić E, Cherwitz D, .Scholes J, Jessurn H, Mc Glennen R

Department of Laboratory Medicine and Pathology, University of Minnesota Medical School, Minneapolis, Minnesota

The polymerase chain reaction (PCR) with polyacrylamide gel electrophoresis was used to study patterns of immunoglobulin heavy chain (IgH) gene rearrangement (GR) in formalin-fixed, parafin-embedded specimens of lymphomas and reactive conditions of mucosa associated lymphoid tissue (MAL,T) and lymph nodes. DNA amplification was performed directly on sections obtained from paraffin blocks. 
Five patterns of PCR products were observed: a single band, two or more discrete bands, smearing, a single band overlying a smear, and two or more bands over a smear. A pure polyclonal pattern (smear) was seen in all of the reactive lymph nodes but in only 15% of cases of Helicobacter pylori (HP) gastritis with lymphoid hyperplasia, 25% of cases of HP gastritis without lymphoid hyperplasia, and 37% of colonic specimens of various types. Patterns consisting of multiple bands with or without background smearing were common in gastritis, colitis, and gastric lymphomas. Single bands or dominant bands were present in all lymph node and salivary gland lymphomas, 12/14 cases of gastric lymphoma and 17/20 cases of 11P gastritis with lymphoid hyperplasia. These bands were reproducible in deeper sections from the same paraffin block or similar areas sampled in difl'erent blocks in all of the lymph node and salivary gland lymphomas, 11/12 gastric lymphomas, but only 1/17 cases of HP gastritis with lymphoid hyperplasia. Bends were also found in 3/20 cases of HP gastritis without lymphoid hyperplasia and 17/38 colonic specimens, but these were not reproducible. 
The complexity of patterns of IgH GR in acquired MALT compared to lymph nodes may be the result of a relative paucity of B-cell clones or preferential proliferation of B-cell clones with a limited area of distribution.

 

HELICOBACTER PYLOR1 (HP) GASTRITIS: WE DID NOT FIND ANY CORRELATION BETWEEN GRADES OF ACTIVITY AND INTENSITY OF MUCOSA COLONISATION BY HP

Gumzej Ž, Meštrić A, Brezovečki D, Belicza M

"Ljudevit Jurak" Clinical Department of Pathology University Hospital "Sestre milosrdnice", Zagreb, Croatia

One hundred twenty-one consecutive antral gastric biopsies showing various grades of colonisation by Helicobacter pylori ( HP ) were studied. Using histochemical and immunohistochemical methods as well as semi - quantitative analyses according to previously published proposals, intensity of HP co(onisation and granulocytic parts among inflammatory cells were analysed. 
Results: There was no significant correlation between density of HP colonisation and neutrophils infiltration. 

 

"SERRATED ADENOMATOSIS COLI" - A NEW TYPE OF ADENOMATOUS POLYPOSIS OF THE COLON

Torlaković E, Snover DC

Department of Laboratory Medicine and Pathology, University of Minnesota Medical School, Minneapolis, Minnesota

Backeround and aims: Hyperplastic polyposis clinica(ly resembles adenomatous polyposis and has not generally been considered precancerous. However, since the original description, a number of cases associated with adenocarcinoma have been reported. The aim of this study was to re-evaluate patients previously diagnosed with hyperplastic polyposis. 
Methods: Pathological analysis of polyps in 6 patients with putative hyperplastic polyposis and 4 with associated carcinoma was compared with classic isolated hyperplastic polyps, adenomas and solitary serrated adenomas. lmmunohistochemical study for the detection of p53 protein, blood groups antigen, including Lewisa and Lewisb and peanut lectin binding was performed. 
Results: Polyps in our patients were much more similar to serrated adenomas than to hyperplastic polyps and were characterised by large size, prominent architectural distortion, cytologically atypical nuclei, focal nuclear crowding and nuclear dispolarity, and rare upper zone mitotic figures. The polyps in our patients and control serrated adenomas had a decrease or absence of endocrine cells compared with classic hyperplastic polyps and normal colon and similar immunohistochemical reactivity for p53 and Lewis' and Lewisbantigens. 
Conclusions: Our results indicate that the polyps in our patients are serrated adenomas. Serrated adenomatous polyposis has not been described before and should be distinguished from true hyperplastic polyposis given a possible association with adenocarcinoma in the former group. 

C-erbB-2 ONCOPROTEIN LEVELS IN SERUM AND TISSUE OF PATIENTS WITH COLORECTAL CARCINOMAS.

Krušlin B, Lukač J, Kovačević D, Kordić D, Čupić H, Kusić Z, Belicza M.

University Hospital "Sestre milosrdnice", Zagreb

We report our preliminary data concerning the determination of c-erbB-2 oncoprotein in senrm and tumours tissue of 38 patients with colorectal carcinomas. The patients were treated by surgery followed by chemotherapy. The serum levets of c-erbB-2 oncoprotein were measured by immunosorbent assay techniques (Oncogene Sciences). lmmunohistochemistry was performed using two different primary antibodies (Oncogene Sciences and DAKO). The alkaline phosphatase/antialkaline phosphatase and horse-radish peroxidase/antiperoxidase methods for investigation of the corresponding tumour tissue was used. lncreased serum c-erbB-2 oncoprotein was found in 23 patients. The extent of serum oncoprotein expression corresponded to the clinical stage of the tumour and to the immunostaining of the tumour tissue. lmmunohistochemical analysis of primary tumours showed strong positivity in one case, moderate positivity in nine cases, and slight positivity in 16 cases. Positive irnnttmostaining was observed in three cases in both primary tumours and lymph node metastasis. Both methods showed comparable results.The serum c-erbB-2 oncoprotein may be useful for monitoring the patients with colorectal carcinomas particularly in cases showing positive immunostaining of primary tumour. Further investigation of a larger series of patients with a long-term follow-up is obviously needed. 
 

REFERENCES

  1. Isola JI et al. Cancer 1994;73:652
  2. Fontana X et al. Anticancer Res 1994;14:2099
  3. Vogel W et al. J Cancer Res Clin Oncol 1996;122:118

B. Iatrogenic, experimental and environmental pathology 
     Iatrogena, eksperimentalna i patologija okoliša

CADMIUM INDUCES MORPHOLOGICAL AND FUNCTIONAL CHANGES OF CELLS IN THE RAT EPIDIDYMIS

Sabolić I, Herak-Kramberger CM, Breton S, Brown D

Institute for Medical Research and Occupational Health, Zagreb, Croatia, and Renal Unit, Massachusetts General Hospital, Boston, USA

The function of epididymis is to accumulate and store spermatozoa, and to generate conditions for their maturation. During their passage through epidydimis, spermatozoa are immobile due to acidic environment in the duct lumen. The respective cells and cellular mechanisms responsible for this acidification have been poorly defined. We have recently indicated clear cells as likely contributors to this process (I). Cadmium (Cd) is a well known pollutant strongly toxic to the male reproductive tract. In experimental animals, subchronic Cd administration causes atrophy of testes and epididymi, impaired spermatogenesis, and loss of acidification in the epididymis (2). In order to further describe the cells that contribute to proton (H~) secretion, and to explain the Cd-induced loss of acidification in the epidydymis, we immunohistochemically characterised organs in control and Cd-intoxicated rats. Four ym thick cryosections of the in vivo fixed tissue were studied by indirect immunofluorescence (3) using monoclonal and polyclona) antibodies to specific subunits of the vacuolar Hč-ATPase (V-ATP-ase), carbonic anhydrase type II (CA lI), and Na/K-ATPase. We confirmed the clear cells as possible H"-secreting cells in the adult rat epididymis. These cells, being high columnar in the head, low columnar in the body, and cuboidal in the tail of epididymis, exhibited a heavily labelled V-ATPase in the apical domain and cytoplasmic localisation of CA Ii. They, however, lacked basolateral staining with the Na/K-ATPase antibody. Following subchronic Cd treatment in rats, all clear cells along the epididymis transformed into high columnar and showed a diffuse cytoplasmic localisation of the V-ATP-ase, whereas the staining pattern with the anti-CA I.I and anti- Na/K-ATPase antibodies remained unchanged. Cells of similar shape and appearance were detected in the epididymi from pre-pubertal rats. We conclude that 
a) clear cells with apically-located V-ATPase are most probably responsible for acidification in the epididymal duct, and b) Cd induces regression of these cells into pre-pubertal (immature) non-secretory form. 
 

References:

  1. Brown D, Lui B, Gluck S, Sabolić I, Am JPhysiol 263: C913-C916, 1992
  2. Calfisch CR and Du Bose TD, Toxicol Environ Health 32:49-57, 1991
  3. Brown D, Lydon J, Mc Laughlin M, Stuart-Tilley A, Tyszkowski R, and Alper S, Hi.stochem Cell Biol 105:261-267, 1996

SELECTIVE EFFECTS OF CADMIUM UPON PLASMA MEMBRANE PROTEINS IN THE RAT PROXIMAL TUBULE

Hernk-Kramberger CM, Thevenori F, Snbolić I

Institute for Medical Research and Occupational Health, Zagreb, Croatia, and II Physiological Institute, University of Saarland, Hamburg/Saar, Germany

Cadmium (Cd) is a potent nephrotoxic heavy metal, which causes severe defects in reabsorption and secretion of various compounds in the mammalian proximal tubule (PT) (for literature see ref (1)). The cellular mechanisms underlying Cd nephrotoxicity are not clear. Possible targets of Cd action at the cellular level could be plasma membrane transporters and enzymes in the PT cells, whose activity may be either directly inhibited by Cd orfand they may be reduced by number (I). It is generaily assumed that Cd-nephropathy resembles the Fanconi syndrome, with an indiscriminative toxicity to all cellular functions. In order to investigate a possible specificity of Cd effects upon plasma membrane proteins in PT cells, by indirect immunofluorescence hisiochemistry (1,2) we studied the expression of several protein in the distinet cell membrane domains using 4 pm thick cryosections of the fixed kidney tissues from control rats and from rats subchronically intoxicated with Cd. Subchronic Cd nephropathy in rats was induced by injecting CdClz (2 mg Cd/kg body wt./day, s.c.) for 2 weeks. Immunostaining of cryosections with monoclonal or polyclonal antibodies to specific plasma membrane proteins revealed; a) the expression of Na/K-ATPase in the basolateral cell membrane was not significantly affected by Cd treatment, b) the expression of several bmsh-border proteins, including vacuolar H'-ATPase, sodium-phosphate cotransporter type II (hlaPi-2), and dipeptidylpeptidase type IV (DPPIV), was dramatically reduced in Cd-intoxicated rats, and c) the expression of multidrug resistance P-glycoprotein (mdrl), a protein also located in the brush-border membrane, was strongly increased by Cd-treatment. Conclusion: in Cd nephrotoxicity the expression of various plasma membrane proteins seems to be selectively affected; the abundance of a specific protein in the plasma membrane can be either unchanged or strongly diminished, or increased by Cd-treatment. The cause of this selectivity is'unknown.

References:

  1.  Herak-Kramberger MC, Spindler B, Biber J, Murer H, and Sabolić I, Pfluegers Arch- Eur J Physiol 432:336-344, 1996
  2. Brown D, Lydon J, McLaughlin M, Stuart-Tilley A, Tyszkowski R, and Alper S., Histochem Cell Biol 105:261-267,1996

GROSS FINDINGS IN ACUTE ATTACK AND LATENT PHASE OF EXPERIMENTAL EPERYTHROZOONOSIS IN PIGLETS

Modrić Z, Grabarević Ž, Milas Z, Robić M

Department of General Pathology and Pathological Morphology of the Veterinary Faculty, Zagreb

Six splenectomized and 12 non-splenectomized piglets, crossbreeds between sweden landrace and yorkshire, with body weights between 12 and 15 kg were inoculated with 2-5 ml of blood from pig which have had more then 90% of erythrocytes infected with Eperythrozoon suis breed Munchen S-261/1990. In non-splenectomized inoculated piglets some time after inoculation splenectomy was performed. Acute attack of eperytrozoonosis in splenectomized inoculated piglets appeared 8-13 days after inoculation and in non-splenectomized inoculated piglets 4-16 days after splenectomy. Microscopic examination of blood smears from piglets in acute phase of disease, stained with Giemsa, revealed in almost every erythrocyte and outside the erythrocytes E. Suis. In disease attack two piglets died and gross examination revealed poor condition, hemolytic icterus, general anemia, hemorrhage and hyperplastic lymphadenitis, parenchyma) and fatty degeneration of liver, enzootic pneumonia, hydropericardium and degeneration of heart muscle, catrrhal inflammation of gastric mucosa with hemorrhages in pylorus, hemorrhagic inflammation of ileum and subdural hemorrhages in brain. In piglets sacrificed in latent phase of disease, 60-171 days after inoculation, interstitial hepatitis and inflammation and dystrophic changes in kidneys, heart, lymph nodes and lungs. 

THE INFLUENCE OF BPC 157 ON NITRIC OXIDE ANTAGONIST AND AGONIST INDUCED LESIONS IN BROILER CHICKS

Tišljar M, Grabarenić Ž, Artuković 13, Robić M, Džaja P, Sikirić P, Seiwerth S

Department of General Pathology and Pathological Morphology of the Veterinary Faculty
Department of Pathology School of Medicine University of Zagreb

The main purpose of this study was to determine the effects of nitric oxide (NO) agonists and antagonists and the influence of a novel organoprotective pentadecapeptide BPC 157, on the development of the broiler pulmonary hypertension syndrome (BPHS) and tissue lesions in the chicks. 
Methods. Acute toxicity. The effects of single dose application of saline (1 ml ip), BPC 157 (10 p/kg b.w.), L-NAME (NO-antagonist/doses 50, 100 and 150 mg/kg b.w.) with their combination (L-NAME+BPC 157, L, NAME+L-arginine/L-arginine/100 mg/kg b.w.) were investigated. Pathohistological examination of the spleen, heart, liver and lungs and haematological analysis were conducted. Chronic toxicity. The chicks were treated daily during five weeks with L-NAME (10 mg/kg b.w.), L-arginine (100 mg/kg b.w.), BPC 157 (10 P/kg b.w.) and their combinations (L-NAME + BPC 157; L-arginine+BPC 157) intraperitoneally (ip). Seven animals from each group, including control (saline, 1 ml ip), were scarified every week. Application of L-NAME caused PBHS in treated chicks. lt was prevented by simultaneous application of L-arginine and BPC 157. Pathohistological examination of both acute and chronic toxicity showed forth a severe tissue damage (myocardial and hepatocyte necrosis, necrosis of lymphoid cell in spleen) caused by L-NAME while haemorrhages, congestion and oedema were predominantly induced by L-arginine. The effects of 1.-NAME was successfully neutralised by the application of L-arginine and BPC 157. The later substance did not cause any visible tissue or organ damage. Haematological analysis shows significant haemoglobin and leukocyte number decrease in the L-NA lE treated group of chicks. 

SCIENTIFIC ESTABLISHED APPROACH TO THE RESEARCH OF THE ENDEMIC UROPATHY

Božić Z

Institution: Private practice, Palmolićeva 45, 10000 Zagreb

Author critically analyses approach to the research of the endemic uropathy (EU), which often lacks generally approved scientific principles and foundations. Significant number of publications and reviews on this subject does not meet elementary criteria expert and scientific informing. Through the past decades in Balkan countries the phenomenon of EU has been burdened with nationalism, marked politicisation , and specially with specific pseudoscientific mystification characteristic for primitive forms of consciousness. First and unavoidable step in research of every disease is establishing its transparent epidemical, clinical and etiologic definition. This scientific principle in the case of AEU has not been respected, which often resulted in inconsistent, chaotic and stochastic research work. This specially reflected on research of the aetiology of disease, which besides evidently pseudoscientific, often assumed party dogmatic and even barlesque character. Many expert meetings on the subject of EU often became its own purpose and exhaled with " 1'art-pour-I'artism ". Great damage to the research of EU was done by forgery of medical documents in endangered rural environment by giving code of disease to persons who were not ill, pushed by charity impulse or organised medical crime. Giving up of eminent world researchers of this field from further i(lumination of EU, persistent recycling of unchanged o1d texts, tables and maps at the recent scientific meetings and powerlessness of state medical institutions in prevention of diseases clearly illustrates dead end street in which this national medical problem has found itself. Author finally calls on abandoning of the spirit of " Balkan tavern " and adaptation of steps in research of EU to standardised scientifically founded principles. 

C. "Herman Jurak" round table on rheumathological diseases 
"Herman Jurak"- okrugli stol o reumatološkim bolestima

MICROVESSEL QUANTITATION IN SYNOVIUM

Djorđević G, Krstulja M, Jonjić N

Department of Pathology, Faculty of Medicine, University of Rijeka, Rijeka, Croatia

Rheumatoid arthritis (RA) is characterised by joint inflammation which results from the tumour-like proliferation of synovial cells as a pannus overlaying and destroying cartilage and bone. The formation of new blood vessels (angiogenesis) is critical in this process. The aim of this study was to analyse the relationship between the microvessel density, detected by inm~unohistochemical analysis using FVIII antigen and proliferative activity of synovial celis determined by nucleolar organiser regions (AgNOR) in three different synovial diseases (RA, bacterial arthritis/BA and osteoarthritis/OA). The microvessel count (the mean number of microvessels in the three areas of highest vascular density at x200 magnification). was significantly hiaher in patients with RA (79.8+30.3) and BA (68.9+37.7) than in those with diagnosis of OA (46.5+21.5), (p<0.05), while the AgNOR parameters was highest in synovial cells of RA (5.5+2.5) than in BA (3.8+1.9) and OA (3.4+2.1) (p<0.0001). 
These preliminary results indicate that vascu(ar proliferation in bacterial arthritis is not accompanied by activation of proliferative markers of synovial cells. In other words signs of increased synoviocytic proliferation appeared to be typical for rheumatoid arthritis. 

D. Ethical forum - Forensic medicine: Profession? Science? Politics? 
     Etički forum - Sudska medicina: Struka? Znanost? PoIitika?

FORENSIC DENTISTRY (PAST, PRESENT, FUTURE)

Kaić Z, Brkić H, Munđušić J

Zavod za deutalnu antropologiju Stomatološkog fakulteta Sveučilišta u Zagrebu

Forensic dentistry can be followed since the days of the ancient Mesopotamia kingdom (Hamurabi 1729 till 1686), till our times. The aims and purpose of the forensic dental expertise are changing throughout the history. This changes are effected by social and legal system as well as by organization of health services in different communities. The forensic dental expertise in our country has begun in the early thirties of this century. The main issues were determination of the damage caused by trauma of teeth and jaws, and qualification and categorization of these injuries. Furthermore, the forensic dentistry dealt with the claims made by patients unsatisfied with their dental treatment. Dental identification of unknown body or human skeleton rests, or the identification of the perpetrator of the human bite mark on the victim, are primary tasks of the forensic dentistry in investigation and accusation procedures. In addition, the measures for improvement of education and practice of forensic dental experts are proposed. Our suggestions are based on the experience of the developed countries as well as on our own, that we gathered during the years in legal trials and in cooperation with police forces, but the major part is based on our experience from the Homeland War (1991-1995) in Croatia. 

UNUSUAL MODE OF DISPOSAL OF DEAD BODY

Marcikić M, Mandić N

Department of Pathology and Forensic Medicine, Clinical Hospital Osijek, Croatia Psychiatric Clinic, Clinical Hospital Osijek, Croatia

Murderers try from time to time to hide crime by disposal of a dead body by very unusual ways. In the presented case accused with murder also apply distinct procedure of disposal of a body by covering it with concrete and by fire. Such kind of disposal could be brutal manifestation of resolving his sexual ambivalence. 

CAUSES OF DEATH OF ROMAN EMPERORS

Šipuš N

Institution: Private practice, Petrova 31, 10000 Zagreb

Tanatalogical analysis has been conducted with 159 rulers of The Roman Empire who had been ruling since 27. year BC until 476. year, when The Western Roman Empire had fallen down. The analysis includes 126 emperors and 33 empresses. Sixty-one of them have died by natural death caused by different diseases, whereas even 95 (59,8 %) of them have died by forcible deaths. Three rulers have died under suspicious circumstances. The forcible death has been caused by fatal accidents (5,3 %), suicides (9,5 %), war injuries (12 %), post trial executions (9%), while most of them (63,5 %) have been killed during overthrows and assassinations in mutual struggle for power. Causes of natural deaths with 61 rulers have been established based on the data from literature, where various symptoms with individual rulers have been precisely described, which made setting a diagnosis possible. 

E. Computer systems aud computerisation in pathology 
     Informatički sustavi i kompjuterizacija u patologiji

COMPARISON OF CLINICAL AND PATHOLOGICAL DIAGNOSIS BY "THANATOS" COMPUTER SYSTEM

Vučić M, Žurić M, Brezovečki D, Periz R, Plasaj T

"Ljudevit Jurak" Clinical Department of Pathology University Hospital "Sestre milosrdnice", Zagreb, Croatia

"Thanatos" computer system for clinical pathology provides memorising and processing of clinical and pathological diagnoses coded according to the International classification of diseases and related health problems (tenth revision, 1994). 
"Thanatos" computer system was used in this work to analyse data originating from the archives of post mortem sections performed in "Ljudevit Jurak" Clinical Department of Pathology during 1995. Main causes of death as well as main clinical conditions were defined according to recommendations of the World Health Organisation published in above mentioned Classification. 
In defined period 1.269 patients died in University Hospital "Sestre Milosrdnice". Their average age at time of death was 67 years, and post mortem section was performed in 548 cases (43% of total). 
By using "Thanatos" computer system three parameters related to patients' death were compared: a) clinical diagnosis, b) main pathological condition and c) pathological cause of death. Comparison was possible in 80,97% of cases, while in remaining 19,03% clinical data were insufficient for the analysis. Among cases which were analysed in 8,75 % all three parameters were in agreement, in 18,16 % clinical diagnoses responded only to main pathological condition, and in 30,64 % clinical diagnoses responded only to pathological cause of death. In a relatively high proportion (42,45 %) of analysed cases it was not possible to record any agreement between compared parameters. 

CAPABILITIES OF "THANATOS" COMPUTER SYSTEM IN CREATING OF SPECIFIC REGISTER FOR THYROID DISEASES

Žurić M, Vučić M, Čupić H, Krušlin B, Belicza M, Periz R, Plasaj T, Zurak K, Ivkić M, Kusić Z

University Hospital "Sestre milosrdnice", Zagreb, Croatia

"Thanatos" computer system for clinical pathology memorises and processes diagnoses coded according to the International Classification of Diseases and Related Health Problems (tenth revision, 1994). We have used mentioned system to build up a specific register for thyroid diseases, and "Thanatos" was expected to fulfil several demands: a) to memorise diagnoses obtained from surgery, b) to differentiate malignant from benign lesions, and c) to differentiate stages of diseases. 
Between 1992 and 1996, 806 patients were submitted to surgery for partial or total thyroidectomy. Out of total, 722 lesions were benign (89,6%), and 84 lesions were malignant (10,4%), giving a proportion of 8,6:1. Benign findings were established as follows: follicular adenoma in 245 cases, nodular goitre 360, diffivse goitre 31, Hashimoto's disease 32, Graves' disease 6, and diffuse hyperplasia 48 cases. Malignancies were classified as follows: well-differentiated carcinoma 71 (papillary, 64 cases and follicular, 7 cases), medullary carcinoma 10, and anaplastic carcinoma 3 cases. 
We have also challenged "Thanatos" system with more complicated task - correlation of the primary tumour size with metastatic rate to local lymph nodes. Model was the most common type - papillary carcinoma. Among metastases free patients primary tumour measured 16,3±12 mm. Lymphatic spread had 31,2% of patients, with tumour diameter of 17,5±11 mm. Difference between the two was not statistically significant, leading to conclusion that size of primary tumour does not predict malignant potential of the papillary thyroid carcinoma. 
In presented work we have demonstrated, on the example of register for thyroid diseases, that "Thanatos" computer system for clinical pathology is reliable in memorising of pathological data and very useful in their analysis, providing valuable scientific data on prevalence and nature of various pathological conditions. 

"THANATOS" PATHOLOGICAL DATA POOL AS A PART OF THE HOSPITAL CANCER REGISTER

Vučić M, Žurić M, Brezovečki D, Belicza M, Periz R, Plasaj T, Kusić Z

"Ljudevit Jurak" Clinical Department of Pathology, Clinical Department of Nuclear and Oncology University Hospital "Sestre Milosrdnice", Zagreb

Pathological diagnoses of biopsies performed during 1995. in "Ljudevit Jurak" Clinical Department of Pathology were coded by "Thanatos" conrpcrter system for clinical patholo~ according to the International classification of diseases and related health problems (tenth revision, 1994). Coding of diagnoses was done on a basis of four digit marks consisting of letters and numbers, and also on a ground of a tumour morphological classification. 
"Thanatos" computer system for clinical pathology provides: a) memorising of specific classifications of malignant diseases, like pTNM system and its variations for different tumours (i.e. classifications according to Clark, Breslow, Dukes, Astler-Coller, etc.); b) grading according to differentiation of various tumours and evaluation of other parameters adjusted to specific malignancies (i.e. Gleason score, mitotic and apoptotic indexes, ete.). 
"Thanatos" computer system was used in this work to estimate a proportion of tumours in total material obtained by biopsies during 1995. Out of a total of 10.604 biopsies, tumours involved 2.779 samples or 26,2%. Malignant findings were detected in 1.494 samples (14,08% of total), and benign lesions were found in 1.285 samples (12,11% of total). Among tumours malignancies made 53,8% of findings, while remaining 46,2% were benign lesions, giving a proportion of 1:1,16 in favour of malignancies. 
Diagnoses memorised in our "Thanatos" computer system are essential for long term follow-up of tumour pathology and they make solid data pool for enforcement of the University Hospital "Sestre Milosrdnice" Cancer Register which tends to become a part of the Croatian National Cancer Register. 

DISTRIBUTION OF THYROID CARCINOMAS ACCORDING TO AGE, SEX AND PATHOLOGICAL DIAGNOSIS - COMPARISON OF TWO FIVE-YEAR PERIODS

Žurić M, Šimunović D, Čupić H, Plasaj T, Kusić Z.

"Ljudevit Jurak" Clinical Department of Pathology University Hospital "Sestre milosrdnice", Zagreb, Croatia

Aim of the study was to investigate possible changes in the prevalence of various types of thyroid carcinomas during two tive-year periods. For that purpose we have reviewed all surgical pathology files between 1987 and 1996. Our interest was limited to the following pathological types of thyroid carcinomas: a) difierentiated thyroid carcinomas /DTC/, (subdivided to papillary /PC/ and follicular carcinoma /FC/); 
b) medullary thyroid carcinoma IMTC/; and c) anaplastic thyroid carcinoma /ATC/. During the period 1987-91. we analysed 88 operated thyroid carcinomas. There were 76 DTC (86,3%), which consisted of 701'C.' (92,1% of DTC, and 79,5% of total) and 6 cases of F(' (7,9% of DTC, and 6,8% of total). Female to male ratio among D'fC was 3:1. MTC was detected in 7 cases (8% of total), with female to male ratio 2,5:1. ATC was found in 5 cases (5,7% of total, 3 females and 2 males). 
(n the period 1992-96, 84 patients underwent surgery due to thyroid malignancy. DTC was diagnosed in 71 cases (84,5%), with 64 T(' (90,1% of DTC, and 76,2% of total), and 7 patients with F'C (8,9% of DTC, and 8,3% of total). Female to male ratio among DTC was 2,7:1. MTC was found in 10 patients (12% of total), with female to male ratio 1:1. ATC had 3 patients (3,5% of total, 1 female and 2 males). 
As presented, prevalence of DTC in both sexes was not significantly changed during two five-year periods. Certain increase of the prevalence of MTC was observed between two compared periods, with large shift in female to male ratio, indicating increase of prevalence among male patients. Finally, relative decrease in prevalence was observed for ATC. 
In conclusion we may say that although no major changes in the prevalence of DTC were observed in the last decade, mild increase of MTC (with lowering of female to male ratio) certainly warrants further epidemiological investigations.