We have great pleasure in inviting you and your colleagues to this symposium
Prof. dr. Križan Čuljak
Head of Department of General Pathology and Pathological Morphology Veterinary Faculty, Zagreb, Heinzelova 55
ORGANIZACIJSKI ODBOR / ORGANIZING COMMITTEE
Prof.dr. Križan Čuljak
Prof.dr. Mladen Belicza
doc.dr. Ruža Sabočanec
|Administrativni tajnik (Administrative secretary):
doc.dr. Željko Grabarevič
Blagajnik (Treasurer): Branka Artuković, dipl. vet.
ZNANSTVENO VIJEĆE / SCIENTIFIC BOARD
Prof.dr. Miro Makek (Switzerland)
Prof.dr. Dušan Ferluga (Slovenia)
Prof.dr. Wieslaw Walkiewicz (Poland)
Prof.dr. Zvonko Kusić (Croatia)
Prof.dr. Karoly Lapis (Hungary)
Prof.dr. Viadimir Mitin (Croatia)
Prof.dr. Mladen Belicza (Croatia)
Prof.dr. Leon Šenk (Slovenia)
Prof.dr. Križan Čuljak (Croatia)
Prof.dr. Ahmed Pirkić (Croatia)
Mjesto održavanja znanstvenog skupa VETERINARSKI FAKULTET, ZAGREB, HEINZELOVA 55
Location VETERINARY FACULTY, UNIVERSITY OF ZAGREB, HEINZELOVA 55
Patološka morfologija humanih i animalnih bolesti
Pathological morphology of human and animal diseases
The terms “fibro-osseous lesions” or “cemento-ossigying fibromas” are confusing and should be discared and replaced by clear and distinct terminology based on clinically, radiologically and histopathologically obsevable and reproducibile characteristics. Histogenetic features and biological potential rather than tumour tissue products alone should be regarded as the basis for nomenclature. A classification and review of clinicopathological features of “fibro-osseous lesions” of tumours origin are presented and illustrated.
Simultaneous multiple primary tumors in head and neck – report of two unusal cases
Nina Gale, V. Kambič, and J. Fischinger
Institute of Pathology, Faculty of Medicine, Ljubljana and Clinic of Otorhinolaryngology and Cervicofacial Surgery, Ljubljana, Slovenia
The occurence of multiple primary neoplasms in the head and neck region is well known phenomenon, the reported incidence being 10-35%. In these cases the second malignant tumor is commonly found in upper aero-digestive trast, lung, esophagus, and less frequently in other unrelated organs.
The aim ofthis report is to present extremely rare appearance of the simultaneous tumors in two patients: the association of the squamous cell carcinoma of the oro and hypopharyngis with the occult papillary adenocarcinoma of the thyroid gland, and the chronic lymphatic leukaemia with the squamous cell carcinoma of the alae nasi. In both patients the coincidental finding of different primary histology in a cervical lymph node was refealed. It represents a vey unusual pathologic finding and only few published reports have mentioned similar coexistence ofthese different malignancies in one lymph node.
Most patients, who developed multiple cancers in the head and neck region die from the second or the third tumor. In case of our first patient the prognosis of the pharyngeal squamous cell carcinoma was worse then that of the papillary thyroid malignancy and the patient die because of the local recurrence of the oro and hypopharyngeal neoplasm. In the secon patient beside the well controlled chronic lymphatic leukaemia and the cured metastatic squamous cell carcinoma of the alae nasi (metastases were found in only two of 25 examined lymph nodes), a fast growing tumor appeared in the upper part of the right lung with multicentric skeletal metastases, most probably due to the third primary neoplasm, However, because the patient refused a bronchoscopy, the tumor was not histologically proven and the patient died three months after the discovery of the lung malignancy.
In conclusion, it is important to emphasize the necessity of an exact, systematic examination of all removed lymph nodes not only to identify the precise extension of the primary metastatic growth but also to uncover a possible second metastatic carcinoma so that appropriate treatment could be performed according to the individual patient.
Prognostic significance of T-cell infiltrates, expression of beta 2microglobuline and HLA-DR antigens in breast carcinoma
Ksenija Lučin, Z. Iternička and Nives Jonjić
Department of Pathotogy, Medical Faculty, 51000 Rijeka, Croatia
It has been suggested that immune mechanisms may influence biological behaviour and prognosis of breast carcinoma. The purpose of this study was to assess the presence of T-cell infiltrates and the expression of beta2-microglobuline and HLA-DR antigens on tumor cells of 75 ductal invasive carcinomas using ABC immunoperoxidase technique. The results were compared with morphometric prognostic index (MPI) that seems to be most accurate prognostic predictor. The extent of T-cell infiltrates differed widely between tumors, but statistically significant correlation was found only with lymph node status, namely tumors with high degree of infiltration had predominantly negative lymph nodes and vice versa (p<0.05). Only 19 (25.3%) out of 75 carcinomas were beta2-microglobuline positive, 34 cases (45.3%) showed heterogenous staining pattern and 22 tumors (29.3%) were completely negative. We could not find any significant correlation between beta2-microglobuline expression and MPI or T-cell content. While normal breast epithelium was always HLA-DR negative, tumor cells displayed positivity in 25 cases (33.3%), 5 tumors (6.7%) were completely positive and 20 tumors (26.7%) displayed only focal positivity. This expression did not correlate with any single prognostic parameter, nor with MPI. The results suggest that T-cell infiltrates and the expression of histocompatibility antigens could not be accepted as prognostic indicators in breast carcinoma.
Adjuvant therapy effect in N(+) breast cancer patients regarding histological grading
A. Bolanča, Z. Čulig, Z. Kusić, C. Lež, M. Belicza
Department ofnuclearmedicine and oncology Universityhospital “Sestre milosrdnice” Zagreb, Vinogradska 29
Twentytwo patients with breast cancer and N(+) receiving postoperative chemotherapy (CMF or PEF) or hormonal therapy (Nolvadex) were devided into three groups according to the histologic grade and followed during the period of two years.
Reccurence of the disease was recorded in 3 out of 8 grade III patients, in 4 out of 6 grade II patients and in none out of grade I patients.
The results obtained demonstrate that histofogical grade can not be used as the only predictor of the disease recurrence in patients receiving postoperative adjuvant therapy. However, these findings should be confirmed on a larger scale of patients.
Association between DNA ploidy pattern and nucleolar regions (AgNORS) in colorectal carcinomas
Nives Jonjić, Senija Eminović, Mira Krstulja, Elvira Mustać and B. Oberman
Department of Pathology, Rijeka University School of Medicine, Rijeka. Croatia
Flow cytometric DNA content analysis is a widely used technique for monitoring cell growth kinetics and tor detecting abnormalities in nuclear DNA content in tumor cells. In colorectal carcinomas it has been concluded that the DNA content represents an independent prognostic predictor. The aim of this study was to find out if there is an association between DNA indexes and the number of nucleolar region associated proteins (AgNOR) in colorectal carcinomas. Seventy-three carcinomas were analysed ior DNA indexes and in 38 cases (52.2~0) aneuploid cell population was found. There was no correlation between DNA indexes and Dukes classification of histopathological differentiation, but the significant differences between AgNOR count and pattems in aneuploid and diploid tumor cells were observed (p<0.05). This result provides the evidence for association between AgNOR and DNA ploidy patterns in colorectal carcinomas, suggesting that AgNOR numbers and pattems can also act as prognostic variables.
Comparative analysis of nucleolar organizer regions (NORs) in human and animal seminomas
D. Baličević, K. Čuljak, Ruža Sabočanec, A. Pirkić, T. Babić, B. Pirkić
Department of Clinical Pathology “Prof. Dr. Ljudevit Jurak”
University Hospital “Sestre milosrdnice”, and Department of General Pathology and Pathological Morphology Veterinary Faculty, Zagreb, Croatia
There are no cytological differences between human and animal testicular pure seminomas in general, butthe biological behaviourof it difters considerably. Seminomas in men are very malignant and tend to metastaze early, but the animal ones are not so agressive. In this sense it may be helpful to examine the regions of the nucleolar organizer (NORs) in the seminoma cells of the both species. Proteins linked to the NORs reflect kinetic of protein synthesis which is markedly increased in a processes of malignancy. The silver staining of NORs (AgNOR) is of considerable diagnostic value concerning both cell proliferation and even malignancy. The number and size of AgNOR bodies varies markedly depending of the grade of tumor difterentiation.
Surgically removed human, dog and parrot testicular tumor masses were histologically verified as a pure seminomas. AgNOR bodies were examined concerning their number, size and shape of the AgNOR bodies (“black dots”) were observed in spite of identical cytological as well as histological appearence of the tumor specimen.
In human seminoma there was a number of predominantly small AgNOR bodies of various shape in an unique cell nucleus. In examined animal seminomas 1-2 rather big dots per nucleus were obvious. This finding implies a possible prognostic speculations concerning the biological behaviour of testicular pure seminomas in various species.
Immunohistochemical demonstration of pituitary peptide hormones in human placentas of various gestational ages
A. Pirkić, B. Hodek, Marija Matković
Department of Clinical Pathology “Prof. Dr. Ljudevit Jurak”,* University Hospital “Sestre milosrdnice”, Zagreb, Croatia
It is well established that human trophoblast is a source of synthesis and/or secretion of a number of peptide hormones among which the role of pituitary-like glandotropins/hormones is not well understood as yet. The aim of our study is to present the immunocytochemical localization of pituitary peptides in the placental trophoblast. Eighteen placentas were examined. Gestational age ranged from l0th to 4lth weeks. Six placentas or placental fragments were from third trimester, four ones from midtrimster, and ten placentas were from third trimester pregnancies which were all of risk. FSH, LH, STH, TSH, ACTH and prolactin were immunohistochemically tested in the fragments of placental tissue after a routine dissection of the placenta.
The results showed marked varieties in the localization and in the intensity of a reaction. In all examined placentas there was an apparent mosaicism of reaction. One group of the villi showed a positive reaction and adjacent ones were negative.
Immunoreactivity of STH was in general intensive and present throughout pregnancy. FSH and LH were much more intensive in thefirst trimester placentas than in placentas from third trimester, where the reaction was much more weaker. Prolactinwas positive solely in several placentas from the last weeks of pregnancy. Reactivity of ACTH and TSH varied markedly, in general it was faint.
The sites of demonstrated pituitary peptides were in a villous trophoblast, in several placentas the reaction was obvious in X-cell of placental bed, as well as in the intervillous syncitial gigantocytes.
The physiological significance of presented findings is rather dubious. Cross reaction, known homology of the pituitary and placental peptides, an “atavism” of the placentamay be a matter of disscusior for the possible explanation. It is very likely that human placenta contains all the components of the hypothalamic – pituitary – adrenal cortex axis.
* and Clinic for Ginecology and Obstetrics
Expression of carcinoembryonic antigen (CEA) in primary ovarian mucinous tumors
Marija Matković, A. Pirkić
Department of Clinical Pathology “Prof. Dr. Ljudevit Jurak”, University Hospital “Sestre milosrdnice”, Zagreb, Croatia
Among all primary common epithelial ovarian tumors the mucinous ones account for 15 – 25%. According to the grade of differentiation they can be divided in benign (mostly cystadenomas with predominantly of endocervical cell type lining), borderline (with predominantly of intestinal cell type) and carcinomas. Many of mucionus tumors show a variety of patterns, so that could be find ina a single tumors abroad spectrum of areas of benign epithelium, as well as areas of borderline malignancies and carcinoma. Distinction between borderline tumors, which are in 10 – 15% invasive, and carcinomas may be difficult. Even 11 % of invasive cystadenocarcinomas show areas of benign appearence. Carcinoembryonic antigen (CEA), an oncofetal antigen, was identified in a variety of normal tissues, as well as in a different malignant lesions. We have examined its expression as a marker for areas of malignant differentiation of benign cystadenomas, borderlinetumors, and malignant ovarian mucinous neoplasias. Immunoreactivity of CEA showed marked variations in localization, it was either apicomembranous or exclusively of cytoplasmic type. In benign mucinous cysts and in benign areas of carcinomas it was negative or membraneous in apical part of the cell. Cytoplasmic type of immunoreactivity was apparent in borderline tumors and in carcinomas, as well as in dedifferentiated areas in benign lesions., where it varied between diffuse and globulogranular type. An intensive granular cytoplasmic type of reaction was evident in the cells scattered in the stroma in areas of tumor invasion.
Occurence of keratoacanthoma in our patients in a two-year period
V. Čajkovac, M. Belicza and Teodora Gregurek-Novak (Zagreb)
Department of Clinical Pathology “Prof. Dr. L,judevit Jurak” and Clinic for Cutaneous and Venereal Diseases, School of Dentistry
University Hospital “Sestre milosrdnice” Zagreb, Vinogradska c. 29
University of Zagreb
The occurence of keratoacanthomas in our patients in a two-year period (19811982) is described. Among total number of biopsies (25004) in this period, keratoacanthoma was diagnosed in 31 patients (11 men, 20 women). In 217 cases of skin biopsies a squamous cell carcinoma was diagnosed.
According to localization the wast majority of the keratoacanthomas (70,98%) was localized on the face, which is an evident proof of the connection with the degree of insoVation. The authors have never noticed its transformation into a squamous cell carcinoma.
Various clinical forms of carcinoma baseocellulare and its incidence in our material
M. Belicza, Teodora Gregurek-Novak, V. Čajkovac
Department of Clinical Pathology “Prof. Dr. !-judevit Jurak” and Clinic for Cutaneous and Venereal Diseases, School of Dentistry
University Hospital “Sestre milosrdnice” Zagreb, Vinogradska c. 29
University of Zagreb
Carcinoma baseocellulare – the most common malignant epithelial tumor shows a great variety of clinical as well as histological patterns. In our material consisting of 164 cases of carcinoma baseocelfulare, according to clinical criteria, on the first place was the solid form and according to histological ones the mixt type predominated.
The frequency of tumors was the greatest in the groups between 50 and 59 ages, while the localization in the majority of cases was od the nose (39,02%) and according to sexes 60,37% of cases was diagnosed in women.
The most important among etiological factors is the ultraviolet radiation which is in a close connection with the localization of tumors on photoexposed areas of the skin.
Vascular tumors of the skin in bioptic specimens
M. Belicza, S. Horvat, Teodora Gregurek-Novak, V. Čajkovac
Department ofClinical Pathology “Prof. Dr. Ljudevit Jurak” and Clinic for Cutaneous and Veneral Diseases, School of Dentistry
University Hospital “Sestre milosrdnice” Zagreb, Vinogradska c. 29
University of Zagreb
Among 2857 skin biopsies in a two-years period (1980-1981) 268 vascular tumors was present and diagnosed. The majority of cases was in age groups from 030 years (34,3%) and 140 patients (52,2%) were female and 128 (47.8%) male. Most common localization of vascular tumors was lower lip (35.7%) chaek (12.6%) and nose (10.5%).
Benign hemangioma with various histological features was found in 217 cases (79.1 %) and angiofibroma in 22 cases (10.1 %). Only one case of sarcoma Kaposi was present (0.4%) and one case of haemangioendothelioma malgnum (0.4%).
Epidemiology and natural history of malignant neoplasms of the urothelial district in the province of Trieste, Italy (1976-1990)
Rossana Bussani, MD, Norita Pavletić, MD and F. Silvestri, MD
Institute of Pathological Anatomy – University of Trieste (Italy) – (Chief: Prof. Furio Silvestri)
One thousand, seven hundred-fifth two new cases of bladder tumors (1331 males, average annual incidence (AAI) 68.42%000, world standardized incidence (WSI) 32.47%000, mean age: 71.8 yrs, and 421 females, AAI 18.63%000, WSI 6.87%000, mean age: 73.5 yrs), and 192 new cases of upper urinary tract (UUl) tumors (134 males and 58 females) were histologically diagnosed among 181,024 biopsies and 40,536 autopsies performed between 1976 to 1990 at the Institute of Pathological Anatomy of the University of Trieste. Over 92% of the bladder and UTT tumors were transitional cell tumors. These last were graded as transitional papillomas in 171 patients, G1 in 290 cases, G2 in 830 and G3 in 499 subjects. Thirty-five percent of cases were invasive cancers (f2, T3, T4). Age-specific incidence rates of urothelial neoplasms increased proportionally to age in both sexes. There was a direct relation between age and high-grade malignancies. Trends in crude incidence rates have shown that, in the period under study, invasive tumors of the bladder significantly increased in females (r=0.933, p<0.001 ). The 5-year actuarial local recurrence rate by specific pathological stage evidenced that in over 55% of cases the tumor recurred. The higher the tumor grade, the lower was the time until local recurrence. The 5-year adjusted survival rate was 67.7% for patients with Ti -neoplasms, 34.4 and 23.4% for those with T2 or >=T3 tumors respectively. Subjects affected by bladder carcinoma had a significantly higher risk of developing a second primary UUT cancer (relative risk (RR): 159.1 for renal pelvis, 277.8 for urethra, 173.9 for ureteral tract;1467.0 person-years) than expected in the general population. As for the extraurinary districts, the sites at highest risk for tumor multiplicity were prostate (RR:16.4, 95% confidence interval (CI): 11.62-22.46), lung (RR: 7.8, 95% CI: 5.31-11.56) and large bowel (RR: 5.1, 95% CI: 2.74-9.49).
Association between bladder and not-urothelial cancers: Incidence and mortality in the province of Trieste (1985-1990)
Luisa Bercich, F. Cavallieri and D. Brunetti
Institute of Morbid Anatomy, University of Trieste; and Tumour Registry of the Province of Trieste, Italy
Incidence and mortality due to multiple cancers of bladder and not-urothelial tissues among residents in the province of Trieste during the 6-year period 19851990, have been studied using the data of the Institute of Morbid Anatomy of the Trieste University and population-based Cancer Registry of the Province of Trieste. Crude annual incidence and mortality rates were calculated by dividing the number of cases by 1,600,873 person-years, that is the sum of estimates of the mid-year populations of the province between 1985 an 1990.
Incidence: There were 146 cases (all histologically verified) recorded (124 in males; 22 in females), giving a crude annual rate of 16.6 per 100,000 in males and 2.6 per 100,000 in females. The cases registered constituted 20.0% of all cases of bladder cancers diagnosed in that period. In males the most frequent associations was between bladder and prostate (50 cases) and lung (31 cases); in females, with lung (7 cases) and breast (5 cases). In 38.4% of the cases bladder cancer developed as first malignancy; the annual incidence of a second cancer among patients affected by bladder cancer was 76.8 per 1,000.
Mortality: Between 1985 and 1990 a total of 56 were recorded (45 males; 11 females); the crude annual mortality rate was 3.5 per 100,000.
Immunohistochemistry of Kaposi sarcoma
C. Lež, M. Bellcza, N. Juranić, M. Balija
Medical Center Zabok and Department of Clinical pathology “Prof. Dr. Ljudevit Jurak” “Sestre milosrdnice”
University Hospital, Zagreb
Two biopsies of cutaneous Kaposi sarcoma in period of six years of 72 years old male patient were studied.
Our aim was to present possible difference in imunohistochemical findings between those two biopsies.
Endothelial markers (ULEX, DAKO 31), collagen IV marker, lympocyte and granuclocyte markers (CD 56, DAKO T4, DAKO L 26, CD 30, DAKO M 1, LCA, suppresor cytotoxic T cell), imunoglobuline and4complement markers (C3 complement, C5 complement, IgA, IgD, IgG, IgE, IgM, kzw:ppa, lambdaj, alpha fetoprotein ad proliferating cells (DAKO Pc) markers were analyzed.
The problem of lung edema syndrome of unknown etiology in a large Pig Farm
Z. Lipejt1, B. Šoštarić2, V. Zidar3,
1Veterinary Institute, Zagreb, Savska cesta 143, 41000 Zagreb
2 Institute forMedical Research and Occupational Health, University of Zagreb, Ksaverska cesta 2, 41000 Zagreb, Croatia
3Pig Producing Farm Dubravica, Dubravica, Croatia
The certain low percentege of pig losses inevitable in every large pig – producing farm are considered as technological ones maintained. The etiology of overwhelming diseases is nor uniform neither specific. In a well managed pig-producing farm in Croatia, over 80% of all losses are examined post mortem systernatized findings are accumulated over years, in order to enable retrospective studies. During the year 1989 an apparently new and fairly uniform pathoanatomical finding was recognized on gross examination of at least 20% of young piglet losser. The problem is still persisting causing similar mortality rate and, inflicting to production considerable economic losses. Epizootiologicaly only the young piglets ranging in age from a few days until 4 or 5 weeks were affected. A peculiar, lung edema with some certain degree of hydrotorax was the only recognizable change. Histopathologic examination of lungs confirmed in every case the gross finding of edema. In some limited number of cases there was a mild interstitial pneumonia, but was considered incidental. Other organs were within normal limits. Numerous bacteriological, virusological, mycological as well as serological examinations of analysed material failed to discover etiological agent. It is believed that new nosologic entity of unexplained etiology appeared, and is persisting on the farm. Recent literature report about lung edema of pigs caused by fumonisin. Fumonisin is a recently discovered mycotoxin produced by Fusaria. It is interesting to note that the problem Qf lung edema in farmed pigs caused by fumotoxin was recognized in USA concurrently with the detection of lung edema of unknown etiology in Croatia.
Primary esophageal melanoma. Case report.
M. Mrčela, B. Dmitrović, M. Marcikić
Zavod za Kliničku patologiju “Prof. Dr. Ljudevit Jurak” Kliničke bolnice “Sestre milosrdnice” Zagreb, Vinogradska c. 29
It is shown a case of 59 years old male patient with primary malignant melanoma of esophagus. Tumor of esophagus was found during gastroscopy and was followed by biopsy. Bioptical Tindings were misdiagnosed as planocellular cancer. After that surgical 2/3 resection of esophagus was performed, and in that bioptical material superficial speding malignant melanoma with metastatic nodul in muscle layer of esophagus was found. Short time after surgical treatment patient died due to pleural empyema. Neither other primary nor metastatic process was found on detailed autopsy examination, and on histological examination of tissue specimens obtained during autopsy. Histological examination of skin changes excluded primary localisation on skin.
We hold this case as a very rare localisation of primary malignant melanoma.
The kidney dysplasia in the autopsy material
Odjel za patologiju, Medicinski centar Bjelovar
The frequency of malformation in the growth of the urogenital tract is considerable. According to available data they make 2% of all autopsies and up to 10% of the autopsies in newborns. The kidney dysplasia is identified by an abnormal differentittion consisting of abnormal collecting small channels, the carthilage metaplasia and metanefric blastema. A normal renal development can be seen as simple glomerules and tubules.
In children most cystic kidneys are dysplastic and vice versa, dysplastic kidneys are usually cystic. The kidney dysplasia is often connected with an obstruction of the urinary tract. This connection as well as interrelation and frequency among kidney malformations on the one hand besides localization and the obstruction rate on the other hand refer us to a theory that displasia in such cases is secondary regarding obstruction of the urinary tract. Instances of this kind of associations are cystic dysplasias with a bad damage of the lower part of the urinary tract and the segmentary dysplasia with a kidney doubling and a syndrome of the ectopic urethrocele.
A connection between obstruction and dysplasia is accepted in theory in some cases, while gardner holds that such a theory has no right to be applicated universally. Different forms of cystic dysplasia arise in several clinical-pathological shapes, but most of them as individual malformations. During two-year period at the Medicat Centre of Koprivnica and Bjelovar 1795 persons died and 752 were autopsied. Of the autopsied there were 109 kidney cysts or 14.5%. The kidney dysplasia was found in two cases of all the cysts or 1.8%. The first case was a male child 3.5 months ofd having a cyst formation instead of a feft kidney with a diameter of 25 mm in which there was pafe-yellow liquid. This formation wighed 15 grammes. The right enlarged kidney weighed 120 grammes and had a lobular surface. On the section the cortex is limited by pyramids without a visible striped drawing. In almost all the pyramids there was a widening of 8 mm containing a clear yellowish liquid connected with the cistern of the kidney. The right ureter is blocked almost completely. The other case was a female newborn of 3 moths. The right kidney weighed 40 grammes and was transformed in a cystic formation with a diameter of 20 mm. The formation was divided by septums in a few cavities with a pale-yellowish liquid. The left kidney, middle-sized, had a lobular surface. On the section the cortex is limited by violet pyramids with a visible striped drawing. Grayish is the mucous membrane of the cistern. The same histological changes were found by histological examenation. There was cartilaginous tissue like small islands, simple and elementary glomerules and tubules. The tubules are partly cystic coated with slablike and low cubic epithelium. There was also connective tissure, immature mesenchyme and in places fatty tissue.
The case of streptococcus pneumoniae pericarditis in guinea pig
Mila Međugorac1, B. Šoštarić1, Borka Šimpraga2
1Institute for Medical Research and Occupational Health, University of Zagreb, Ksaveiska cesta 2, 41000 Zagreb, Croatia
2Veterinary Institute Zagreb, Savska cesta 143, 41000 Zagreb, Croatia
In one small guinea pig colony raised for laboratory investigations an outbreak of high mortality occurred, and several animals died within short period of time without notified clinical signs of illnes. During that period the animals were not subjected to any type of investigation. On autopsy the major lesion were located within the chest cavity, which contained a few ml. of hemorrhagic fluid. The pericardium was thickened by yelowish masses of fibrin. When opened, from pericaldial sac, turgid fluid with fibrin particles were obtained. The epicardium was embedded in thick layers of fibrin and no adhesions were noted. The lungs were hyperemic and edematous, but no pneumonic changes were present. On histopathological examinations the pericardium and epicardium are markedly thickened by diffuse infiltration of inflammatory cells and layers of fibrin. The mesothelial membranes are completely affected, and there is frequent bridging of the epicardial-pericardial space by masses of inflammatory cells and other reactive inflammatory components. The inflammatory reaction consists of fibrinopurulent, almost pure neutrophil, regions, and regions with predominantly reactive fibroplasia with a prominent histiocitic component. There is mild attenuation of the sub-epicardial myocytes with interstitial inflammatory cell infiltrate. At high power, cocci, diplococci and paired coccobacilli are seen. Special stain demonstrated Gram-positive cocci and coccobacilli, usually in pairs. By bacteriological culture on blood-agar media a small beta hemolitic colonies, of paired, capsulated cocci were cultivated. The condition was diagnosed as Streptococus pericarditis.
Incidental malignant tumours in 40-years period
H. Čupić, M. Belicza, Ž. Gumzej, I. Petric, D. August
Department of clinical pathology “Ljudevit Jurak” “Sestre milosrdnice” University hospital, Zagreb
The frequent source for mortality and epidemiological cancer statistics are death certificates. This is the main reason why malignancies are generally underestimated.
Autopsy detection of clinical unrecognised malignancy is relatively common, therefore autopsies are of great value in the control of mortality statistics of a population.
Purpose of our research was to evaluate extent of incidental malignant tumours, which were considered as a cancer found incidentally at autopsy and which could have been found intravitally if a proper diagnostic procedure have been utilized.
We retrospectively studied 27875 autopsy records from autopsies performed at Department of clinical pathology “Ljudevit Jurak” in University Hospital “Sestre milosrdnice”, Zagreb, between January 1953. and December 1992. Incidental malignant tumours were found in 3,24 percent of autopsies performed in studied period with lungs and stomach as leading organs. The age distribution showed a peak incidence in the eight decade, when morbidity and mortality were usually associated with more than one disease process.
Histological study of the joint cartilage degeneration in the knee
Dora Brezovečki-Biđin1 and M. Pećina2
1Institute of Clinical Pathology “Dr. Ljudevit Jurak” Clinical Hospital “Sestre milosrdnice”Zagreb
2 Department of Orthopaedic Surgery School of Medicine University of Zagreb
Although clinical and radiological classifications of the osteoarthritis of the knee joint is generally accepted, the histological stages of the degenerative changes of the cartilage are still under investigation. Especially interesting is the study of comparison of clinical, radiological and histological stages. The aim of this paper is a comparative analysis of histological changes of the cartilage relative to clinical and radiological stages of the gonarthrosis.
In 34 patients who had undergone surgery for gonarthrosis a sample of degeneratively changed cartilage was taken for histological examination. The patients had been evaluated clinically and radiologically before surgery as to the progression of arthritic changes in the knee.
The comparison of clinical, radiological and histologicaf results showed it most appropriate to rate arthritic changes of the knee in three stages- and earfy stage, a moderately advanced stage and an advanced stage.
Comparative analysis revealed that in the first or early stage of the degenerative changes of the cartilage radiological and histological results were more similar, while in the second stage of the changes ciinical and histological results were more similar than those obtained by radiological and histological examination. The same can said for the degenerative changes of knee joint cartilage. Our research confirmed the existence of the so called clinically mute gonarthrosis.
It has also shown that the standard histological method with colouring hemalauneozine is sufficient to enable visual differentiating of the progression of the degenerative changes in the knee joint cartilage. When biopsy allowed only small piece of knee cartilage which didn’t contain all the strata of the cartilage, application of PAS reaction was found to be useful. fVamely, in a normal joint cartilage there is a ring of PAS positive substance around each cartilage cell, whereas in a degeneratively changed cartilage PAS positive substance in the form of a ring appears only around the batch of cells.
The result of comparative studies indicate that a histological differentiation of degenerative changes in the joint cartifage can be applied both in clinical subjective and objective symptomatology and in radiologically visible changes of the arthrotic knee.
Iatrogena patologija i patologija okoliša
Iarogenic and enviromental pathology
Leptospiroza: profesionalna bolest pri radu s laboratorijskim miševima
Z. Modrić, D. Vukelić, Alenka Ballarin-Perharić, K. Čuljak, Višnja Škerk i Višnja Šarić
Zavod za mikrobiologiju i zarazne bolesti, Ileterinarski fakultet Sveučilišta u Zagrebu; i Klinika za infektivne bolesti “Dr Fran Mihaljević” Zagreb
Infekcije leptospirama u ljudi od laboratorijskih miševa ustanovljene su prvo u Amsterdamu 1949. god., a zatim u SAD i Češkoj. Kod nas smo prvu infekciju leptospirama u ljudi od laboratorijskih miševa ustanovili 1986. god. Od leptospiroze oboljele su dvije osobe (jedna 1986. i druga 1988.) od kojih je jedna radila u uzgoju laboratorijskih miševa, a druga s njima u laboratoriju, U obje osobe serološki je potvrdena dijagnoza leptospiroze, a u jedne od njih leptospire su izdvojene iz krvi. Iz uzgoja je na leptospirozu pretraženo 145 miševa u kojih su u 75,17% seruma ustanovljena protutijela za leptospire serološke varijante sejroe, saxkoebing i hardjo i izdvojene su leptospire iz bubrega uzgojem u Korthofovoj podlozi u 73,80% miševa. Za izdvojeni soj leptospira iz krvi pacijenta i za izdvojene sojeve iz miševa ustanovljeno je u reakciji s antiserumima leptospira da se radi o serološkoj varijanti sejroe. Od deset drugih osoba koje rade u uzgoju miševa ustanovljena su protutijela za iste serološke varijante leptospira u još tri osobe. Na osnovi epidemioloških podataka, nalaza protutijela za leptospire u ljudi i miševa i determinacije izdvojenih sojeva leptospira iz oboljele osobe i miševa, smatramo da su laboratorijski miševi bili izvor infekcije za Ijude.
Sinonasal adenocarcinoma in a carpenter
1Ž. Ferenčić, 2M. Ivkić, 1I. Petric, 2V. Petric
1Department of Clinical Pathology “Ljudevit Jurak” and
2Department of Otorhinolaryngology and Head and Neck Surgery, Clinical Hospital “Sestre milosrdnice” Zagreb, Croatia
A 77-year – ofd male carpenter complained of a 2-month history of persistent nasal obstruction, nasal bleeding ad headache. The patient was admitted to the hospital with suspicion for nasal polyps. Computerized tomography revealed a nasal mass exteding to the right maxillary and ethmoidal sinus with intraorbital extension. Probatory right nasal biopsy diagnosed the tumor as a necrotic carcinoma. A right maxillectomy and ethmoidectomy extending to the right frontal and sphenoidal sinus with right orbital exenteration was performed. The tumor after histopathologic examination was diagnosed as an adenocarcinoma of intestinal type. Immunohistochemically the tumor expressed carcinoembrional antigen and cytokeratin positivity, with chromogranin-A, neuron-specific enolase, and S-100 protein negativity. Surgical excision was followed by radiation therapy (6000 cGy), and a 3- month follow-up indicated there was no recun-ence of the neoplasm. The patient had a history of exposure to wood dust for 46 years, and to polish and nitrous solvents for two years working in a furniture factory. This is a first case of sinonasal adenocarcinoma of intestinal type with an association to occupational carcinogens, studied immunohistochemically at our Institution.
Hystological and cytological changes of hyperactive thyroid gland treated with thiamazole and radioactive therapy
Departmenf of Pathology, Medical Center Zadar, Zadar, Croatia
In this study systematical cemiquantitative analysis was done of particular morphological markers typical of diffuse hyperthyreosis and hystological comparison too. Cytologicaly the following morphological features were analysed: hyperplasia (i. e., cellularityof smear), anisomacronucleosis, honeycombed structureofcytoplasme, and admixture of lymphocytes in smear. Hyperplasia and granules in cytoplasme significantly more common were found in the group of thiamazole treated patients without remission than in the untreated group. There was no statistically difference between the other parametars observed. C~uantity of lymphocytes was significantly more abundadnt in the group of patients with no remission than in controll group. Hystological analysis include hyperplasia, anisomacronucleqsis, fibrosis and lymphocitic infiltration of thyroid gland. Statistically significant difference was established for hyperplasia and anisomacronucleosis except for fibrosis. Quantity of lymphocytes was significantly more detected in the group of patients with no remission in correllation with group of euthyreotic patients. Comparison between cytological and hystoiogical material revealed that morphological features “hyperplasia” and “anisomacronucleosis” were constantly stronger expressed in the group of hyperthyreosis with no remission during thiamazole therapy in correllation with group of untreated patients. Especially these cytomorphological changes were expressed in goiters during radioactive therapy. All these cytomorphological benign features of hyperactivity in material examed were caused by continous TSH stimullation during therapy or were caused by exacerbation of leading disease.
Is the “cobweb” material distributed over Croatia during the war a possible threat to human health?
B. Šoštarić1, R. Fuchs1, Mercedes Wrischer2
1Institute for Medical Research and Occupational Health, University of Zagreb, Ksaverska cesta 2, 41000 Zagreb, Croatia
2lnstitute “Ruđer Bošković”, Bijenička cesta 54, 41000 Zagreb, Croatia
During the aggression 1991-1992 against the Republic of Croatia, Yugoslav Federal Army air forces applied among other materials of insufficiently explained purpose a cobweb-resembling material. The first documented attack with material in question was recorded at the end of September 1991 near the town of Sinj. Since then until the end of 1992, 200 well documented droppings of the same material were recorded throughout entire Croatia, encompassing virtually all large towns. There are numerous reports about distribution of the same material over the Republic of Bosnia and Herzegovina. The material was distributed in all occasions from enemy airplanes. It is interesting that in both countries the “cobweb” was distributed over the areas inhabited by the Croatian and Moslem population, while so called Krainas inhabited by the Serbs were not exposed. Reaching the ground the material becomes disavowed from snow flake – like – tufts into very thin white filaments covering in easily perceptible concentrations 2-3 square kilometers. No immediate harmful effects over human or animal population were noted in any case. By toxicological and microbiological analysis nor toxin or any microorganism were detected by several laboratory methods applied. On morphological examination, using scanning and transmission electrone microscopy, two basic type of fibers were determined, both of surprisingly small and uniform diameter. A thicker was, 0.4- in diameter without affinity for heavy metal salts, and the thinner, 0.1- in diameter with marked affinity for heavy metal salts. Chemical analysis showed that thinner fibers are composed of at least 7 amino acids. Apparently it is a first use of this material in the world, with largely unknown purpose. However it is obviously a product of high technology, and further research is needed to explain whether this material might have a harmful eftect on human health when present in environment for a long time.
The outbreak of latrogenic intoxication of one horse herd by dyphteria toxin
B. Šoštarić1,Z. Lipej2, P. Stanković3, Borka Šimpraga2, R. Fuchs1
1Institute for Medical Research and Occupational Health University of Zagreb, Ksaverska cesta 2, 41000 Zagreb, Croatia
2Veterinary Institute Zagreb, Savska cesta 143
3Institute of Immunology, Department for Antisera and Laboratory Animal Production, Svetonedjeljska cesta 15, Brezje, 41431 Sveta Nedjelja, Croatia
During the end of September 1992 in a large hiperimmune antisera producing facility in Croatia 13 horses out of 20 animals in the herd serving for diphteria hiperimmune serum production died suddenly some 36 hours after receiving subcutaneous injection 0.5 ml of 140 Lf-s toxin of diphteria. The remaining 7 horses presenting severe clinical signs of sickenees were treated with a large doses of hiperimmune serum for diphteria. Despite therapy one animal died some 9 days later, while the remaining 6 animals survived, but because of poore recuperation progress were utilized for meat several weeks later. Namely the horses were at the very beginning of production period, and instead of anatoxin they by mistake received dyphteria toxin. On gross pathological examination of all acutely died animals there was a large subcutaneous edema at the place of application. To summarize, generalized serohemorrhagic lymphadenitis with widespread hemorrhages throughout entire body economy were the most striking finding. Degeneration of myocardium, kidney and liver were constant in all cases, and quite severe. Histopathological examination revealed difuse degeneration and necrosis of myocardium, characteristically involving Purkinja-s cells. Acute tubulonephrosis, involving mostly proximal tubule epithelium with isolated cell calcification was pronounced. There was difuse necrosis of germinal centers in lymphoid folicles of spleen and lymph nodes. The liver exibited micro and macrovacuolar degenerative changes.
The tissues of animal succumbing 9 days after appiication presented generaly same type of lesions, but with obvious reparative processes. So in the myocardium a large areas of fibroblast proliferation with detectable colagen formation were present.
All recognized lesions were typical for dyphteria toxin intoxication, and resembled very much pathologic changes in fatal human cases. It has to be pointed out that horse is extremely suceptible for dyphteria toxin.
R. Fuchs, B. Šoštarić, Božica Radić
Institute for Medical Research and Occupational Health University of Zagreb, Ksaverska cesta 2, 41000 Zagreb, Croatia
Ocratoxin A, a nephrotoxic mycotoxin is a secondary metabolite produced by certain Aspergillus and Penicillium spp. The intoxication with this toxin are well documented worldwide in different species of domestic animals, and even more there is accumulated evidence that ochratoxin might play a crucial role in the etiology of Balkan Endemic Nephrophaty. In this study the distribution of radioactive C-14 labeled ochratoxin was investigated in mouse, rainbow trout and Japanese guail. To the animals of every mentioned species radioactive C-14 labeled ochratoxin was injected intravenously, corresponding to 200, 160 and 70 ng/g body weight, respectivety. After difterent survival times the animals were anesthetized to death with CO2 and obtained frozen sections of whole bodies were studied autoradiographically. The samples of certain target organs from frozen carcasses were collected and subjected to chemical analyses. In the mice, the most striking feature was the retention of radioactivity in the blood, which was high even 4 days after application. In the kidney concentration was only a half or one fourth when compared with blood. Contrary, the blood concentration in the rainbow trout decreased to one tenth of the concentrations in the kidney and the bladder few minutes after injection. In the Japanese guail the blood concentration of ochratoxin A remained higher during a longer period than in the trots, but not as long as in the mice. Some 6 hours after application the concentration in liver, kidney and intestinal content was higher than in the blood. In the eggs which were developing, a rather high retention of radioactivity was detected 24 hours after application.
Studies on hepatotoxicity of potassium clavulanate
A. Lavrić, L. Šenk* Z. Kopitar and Doris Tibaut
Lek, Pharmaceutical and Chemical Companyd.d., Research and Development, Celovška 135, 61000 Ljubljana, Slovenia
*Veterinary Faculty, Gerbičeva 60, 61000 Ljubljana, Slovenia
We conducted a study on the acute and sub-chronic toxicity of potassium clavulanate to mice and rats. LD50 in both sexes of the mice amounted to approx. 10.000 mg/kg and >10.000 mglkg in rats. Sub-chronic toxicity (90 days) at the highest dose level (490 mg/kg/day) showed a rise in LDH, but there were no other substantial changes. What we were interested in was to establish what changes occurred in the liver of the rats when those were treated with extremely high doses of potassium clavulanate for several days on. We supposed that an accumulation of the substance in the organs and tissues should take place. Thus rats of bought sexes were treated for the period of 4 days by oral application of 1000 and 5000 mg/kg/day. A daily dosage was divided into three equal parts and supplied every eight hours. After the fourth day of application, blood samples af the animals were taken for a biochemical examination and for the determination of the active substance concentration in the plasma. The animals were then sacrificed in other narcosis. Patomorphologic changes were described. Liver was weighed and liver samples were taken for pathohistologic examinations. It was established that the active substance after a 4 day treatment at dose levels of 5000 mg/kg/day caused a substantial decrease in body weight, moderate gastroenteritis and a marked reduction in liver weight. On the basis of biochemical studies (AST, ALT, LDH) it could be observed that at high doses of potassium clavulanate there was an enryme increase tendency which was confirmed also by pathohistologic examinations (hydrops degeneration presetn). Nevertheless, it is important to stress that those doses were extremely high (1000 and 5000 mg/kg/day) compared to the highest therapeutic dose which was approximately 6 mg/kg/day.
The influence of hyperthermia and cyclophosphamide on the growth of akr leukaemia in mice
M. Radačić, J. Lukenda, Višnja Šverko, J. Jerčić, M. Boranić
Ruder Bošković Institute, Department of Experimental Biology and Medicine, Zagreb, Croatia and Veterinary Faculty, University of Zagreb, Zagreb, Croatia
In last two decades hypertermia as single treament and combined with cytostatics has acguired a wide range of use in experimental models and human tumours. We have investigated the infl~tence of hyperthermia and cyclophosphamide (CT~ on the growth of AKR leukacmia. AKR mice were injected intravenously with 106 leukaemic cells. CTX (100 mg/kg) was given intraperitoneally at different times before and after hypertermia, which was given as whofe body hyperthermia (WBH) in water bath (40.5°C – 42.5°C). Neither CTX nor WBH as a single treatment have had any influence on tumour growth, but if combined treatment was applied, antitumour effect was obtained, The best antitumour effect was achieved when CTX was given 4, 2,1 or 1 / 4 hours before WBH, and was the highest if CTX was given 1/4 h before WBH.
The effect of cisdiamminedichloroplatinum and hyperthermia on tumour growth and kidney function in mice
M. Radačić, Višnja Šverko, J. Lukenda, J. Jerčić
Ruđer Bošković tnstitute, Department of Experimental Biology and Medicine, Zagreb, Croatia
Cisdiamminedichloroplatinum (cis-DDP) is an antitumour agent widely used in the treatment of malignant diseases is restrected due to its nephrotoxic effect. Both antitumour and nephrotoxic effect of cis-DDP is enhanced when it is used with hyperthermia. The enhanced effects are time dependent, i.e. the lower the time interval between aplication of hypertermia and cis-DDP, the better the a~titumour effect and the higher the kidney damage. CBA/HZgr female mice were injected into the footpad with mammary carcinoma cells (106). When the tumour volume was 150200 mm3, tumour bearing mice weretreated with hyperthermia (40.0°C – 43.5°C), cisDDP (2-10 mg/kg), or hyperthermia and cis-DDP combined. Cis-DDP was given intraperitonealy, and hyperthermia was given locally in a thermostated water-both. The obtained data shows that cisDDP as a single treatment does not influence tumour growth, while hyperthermia has a slight antitumour effect. When used simultaneously, the antitumour eftect as well as the toxic effects are highly onhanced.
The influence of nonspecific immunisation of the gilts with Baypamun® on the passive immunity and production parameters of the piglets after weaning
V. Zidar, M. Laušin, Nada Vijtiuk, D. Radeljević, V. Bilić, B. Krsnik, I. Vrbanac and I. Valpotić
Department of Biology, Veterinary Faculty, University of Zagreb, Zagreb, Croatia
Biologic response modifier Baypamun obtained trom inactivated Parapox ovis virus (strain D 1701) was tested in order to establish whether or not it could (1) stimulate lacteal immunity in first-litter gilts, (2) enhance passively acquired humoral immunity in their sucklings, and (3) decrease losses due to infections such as gastrointestinal syndrome (GIS) among pigs during preweaning period. Ten partiurent gilts were given I/M 2 ml of BaypamunŇ solution 9, 7 and 2 days before anticipated farrowing. Two control groups comprising 10 partiurent gilts or sows each received 2 ml of saline as a placebo. The performance parameters of their offspring were recorded as well as clinical, bacteriological, immunological and ambiental observations were made in the farrowing unit of a swine farm in northwest Croatia. Serum levels of immunoglobulins and total proteins in pigs bom to gilts injected with Baypamun® were significantly higher 7 days (P<0.01) and 14 days (P<0.01; <0.05) after farrowing respectively, than those in both control groups. Pigs bom to Baypamun® – treated gilts survived much better (8.6/9.5 or 90.5% weaned pigs per (itter) than did offspring from control sows (7.9/9.9 or 79.8%) during the preweaning period. No losses due to G IS were observed in principals. Fifty percent (in litters of sows) or 83.3% (in litters of gilts) of total losses were diagnosed as GIS among control pigs.
Prosuđivanje starosti procesa kod mikobakterioze svinja na osnovu eksperimentalne infekcije s mycobacterium avium / mycobacterium intracellulare
L. Šenk, M. Pogačnik
Veterinarska fakulteta, Ljubljana, Slovenija
Proučavali smo starost tuberkuloznog procesa u limfiim čvorovima 12 svinja koje su bile per os inficirane sa mikobakterijama iz kompleksa M. avium/M. intracellulare in 1 svinje, inficirane kontaktom. Posle aplikacije 120-140 miliona mikobakterija kroz tri dana ustanovili smo prve histološke tuberkulozne promjene u limfnim čvorovima 91. dan posle infekcije (p.i), a makroskopsko 104. dan. Kazeoznu nekrozu smo ustanovili 104. dan, tog dana i prve znakove inkalcinacije, a očitu veznotkivnu demarkaciju 130. dan p. i. Prve morfološke promjene se manifestiraju u akumulaciji makrofaga i epiteloidnih stanica te u pojavi gigantocita, dakle sa slikom proliferativne tuberkuloze infiltrativnog tipa. Nakon toga brzo uslijedi nekroza tih stanica i nastanak najprije unicentričnih inkapsulamih nekrotičnih žarišta, a posle razvoj njihovih konglomerata i konačno nastanak multicentričnih žarišta. Gigantocite smo našli 130. dan, a resorptivne tuberkle oko inkapsuliranih nekrotičnih žarišta već 104. dan p. i. Primarnog afekta u tonzilama i tankom crijevu nismo ustanovili ni makroskopsko ni mikroskopsko. Kod svinje koja je bila u drugom pokusu izložena kontaktnoj infekciji, ustanovili smo prve tuberkulozne promjene već kod klanja 104. dan p.i.
BPC 157 in incisional wounds healing
S. Seiwerth, Sanja Đačić, P. Sikirić
CDD, Institutes of pafhology and pharmacology Medical Faculty, University of Zagreb, Croatia
Different peptides with large-scale effects, such as PDGF, FGF, EGF, also possitivelly influence wound healing(1 ). Recently, a new pentadekapeptide BPC 157, showing broad organoprotective action, was synthetised (2). In our experiments we evaluated the influence of BPC 157 on incisional skin wounds in rats. Material and methods. In our experiments we used albino rats of Wistar and Fisher strain,180-210 g b.w., either sex. 3 cm incisions through the skin were placed on the back of the animals. On the side of each incision 0.2 ml of either BPC 157 or saline were applied. BPC 157 was applied in a dose of 0.5 ng, 0.5ng or 0.5 per wound. Histological appearance and tensile strenght of wounds were evaluated after 3, 5, 7, 14 and 21 days. Statistical analysis was performed by Mann-Whitney test. Results. Histologically, a significant difference between controls and BPC 157 treated animals was observed conceming edema, inflammatory celfs population and collagen presence. Likewise, tensiometry showed signifficantly better wounds healing dynamics in BPC 157 treated animals than in controls. Conclusion. From our results we conclude that BPC 157 represents a perspectivelly potent tool also in human wounds managing.
1. T. A. Mustoe et al. Science 1987; 237:1333.
2. P. Sikirić et al. Acta Physiol Hung 1992; 80:89.
Domperidone and restraint stress induced pancreatic beta cells lesions in rats
Sanja Đačić, Ž. Danilović, Ž. Grabarević, I. Rotkvić, M. Devčić
Institute of pathology, Medical faculty University of Zagreb, Department of pathology, Veterinary Faculty of Zagreb
The aim of this study was to investigate the interaction of the dopamine system and stress on pathological and biochemical changes of the endocrine pancreas in rats. Wistar Albino rats were treated with domperidone (0.2, 2.0, 20.0 mg/kg b.w. i.p.) or saline (5 ml/kg b.w. i.p.) 1 hour prior to restraint stress. We applied domperidone in the same doses to animals not exposed to restraint stress. The animals were sacrificed after 1, 2, 3, 4, 5, 6, and 24 hours. Serum insulin and C-peptide concentrations were determined (RIA method). The pancreas was analyzed using histological and immunohistochemical methods. Domperidone’s effect on the endocrine pancreas during stress can be achieved only in dose of 20 mg/kg b.w. especially in the first hours of stress and delays the organism’s adaptation to stress.
1. Pare WP, Glavin BG Restraint stress in biomedical research. Neurosi Biobehav Rev 1986; 10:339-70.
Comparison between stomach lesions induced by short-time pyiorus ligation method and original Shay method
Ž. Grabarević1, I. Rotkvić2, M. Dodig3, V. Šimičević4
1Department of Pathology Veterinary Faculty Zagreb,
2Departmenf of Gastroenterology Clinical hospital “Sestre milosrdnice” Zagreb,
3PLIVA Pharmaceutical Company Zagreb
4Medical Faculty University of Zagreb
Pylorus ligation was originally described by Shay et al. in 1945. as a simple and reproducible method for investigations of gastric lesions and secretion. Although ther focused their work on the stomach lesions developed after 17 till 19 hours after pylorus ligation, they also described lesions which were developed only after 4 or 6 hours of ligation. Today, the routine use of this method applies pylorus ligation which lasts 6 hours. Using the original Shay method no gastric lesions were observed after 30 min or 1 hour of ligation. However, in this work we described haemorhagic stomach lesions developed after very short time (only 15 min) of ligation. The main difterence is the exact position of the ligature. Macroscopically and histopathologically these lesions are identical to Shay’s developed after 4 or 6 hours of ligation.
Morphological and functional changes of the incapsulated Langerhans islets before and after transplantation
M. Hadžija, Marina Ćetković-Cvrlje, M. Slijepčević, Mirjana Gavella*, Vaskresenija Lipovao*, Đ. Vranešić
Ruđer Bošković, University of Zagreb, Croatia,
*Vuk Vrhovac Institute, Univeisity of Zagreb, Croatia,
New Hospital, University of Zagreb,
Various antigen present on Langerhans islets were suggested to be involved in autoimmune destruction of pancreas in diabetes. Ganglioside, a sialic acid containing glycolipid, was also suggested to be one of the target antigens. Microcapsules may protect islets transplanted, either in humans or animal models of diabetes, from transplantation reaction or autoimmune destruction. We evaluated whether HEMAMMA-polyacrilamide microcapsule protects islet cell insulin secretory capacity in relation to cell sialic acid content. We compared insulin secretory response (m U/L.i./ min) and surface sialic acid content (ng/L.i) on isolated and encapsulated islets: 1) freshly isolated and encapsulated islets, and 2) tissue culture – free and encapsulated islets maintain for seven days under optimal conditions (5% C02 37°C, RPM) 1640+10% FCS). Best time for islets encapsulation was after 24 h in tissue culture (insulin secretion 16.O± 1.7) of islets. Insufin secretion was retained after 2nd (6.2± 0.6) and 4th (12.4±1.2) day and diminshed from day 7th (6.2± 0,8), An increased concentration of sialic acid of encapsulated islets, 2nd (21.2± 3.8), 4th day (45.8± 5.4), 7th day (69.3±7.3) was observed in tissue culture. Preliminary results of peritoneal implantation of HEMA-MMA-polyacrylamide microencapsulated islets effectively reduced blood glucose in xenogenic combination. After extirpation of microcapsules in vitro, insulin secretion was slightly reduced (20%) compared to values of freshly encapsulated islets, and sialic acid concentration was reduced by 16% under same conditions. In conclusion, these results suggest that a period of four days of tissue culture is the best time for optimal quality of HEMA-MMA-microcapsule implantation.
Sudska medicina: Profesija? Znanost? Politika?
Forensic medicine: Profession? Science? Politics?
The problems of war victims identification
D. Strinović, J. Škavić, D. Zečević, S. Gusić, M. Kubat, J. Čadež, Marija Definis (Zagreb)
Department of Forensic Medicine, Zagreb
University School of Medicine, Zagreb, Croatia
The intention of this paper is to describe the organizational principle and indicate the results already achieved in the identification of war victims in Croatia. 6.493 victims had been identified by February 25th i 993. A model is proposed that could be used in the course of identification processes, examining the methods and principles of identification, complicated by the time interval or more than a year from the time of death, for a presumed number of several thousand (up to 14.000) unidentified victims, possibly in mass graves. Identification will be further complicated by the lack of antemortem medical and dental records and the incapacity to utilize more expensive methods of identification. Attentions draw to group of more complex cases examined at the Institute of Frencis Medicine and Criminology.
The problem differential diagnosis of the shoot wounds induced with fast projectils
J. Škavić, D. Strinović, D. Zečević, S. Gusić, M. Kubat, J. Čadež, Marija Definis-Gojanović
Department of Forensic Medicine, Zagreb University School of Medicine Zagreb, Croatia
Gunshot and explosive wounds were the most frequent causes of death in the war against Croatia during 1991-1992. For some of them, even after the autopsies, it wasn’t possible to conclude if they were inflicted by projectiles or by fragments from explosive devices. Several examples are presented in this paper. The artical calls attention in interpretation of the mechanism of wounding because these wounds sometimes have a great morfological similarity, as well as they could resemble the other types of wounds or injuries (eg.incised wounds, blunt force injures).
Forensic medicine activity problems in Split region since war beginning in RH and BiH
Marija Definis-Gojanović, Š. Anđelinović, J. Ivanović
Department of Pathology and Forensic Medicine, Clinical Hospital Split, Split, Croatia
The War imposed to the Republic of Croatia has brought to the Department of Pathology and Forensic Medicine, Clinical Hospital Split, new taske as well as many problems, most of them mainly out of the medicine profession. The authors carry out their experiences and difficulties they have met trying, with their best, to treat and equip dead bodies: members of Croatian army and civilians, as well as dead members of aggressor’s formations. The problems are increasing by flaming the war in the Republic of Bosnia and Herzegovina. From the beginning there was no interest and so no cooperation with communalis, civilians and militaries judical organisation of ours, as well as neighbouring republic, which we have been persistently trying to reestablish. Til( today, the condition hasn’t been changed in principal because of widely desorganization of system of authorities and responsibilities.
Some ethical aspects of hyperimmune sera production on horses
D. Stanković1, B. Šoštarić2, N. Petrinec1, Astrida Heisinger-Purić1, J. Petek1
1Institute of Immunology, Department for Antisera and Laboratory Animal Production, Svetonedjeljska cesta 15, Brezje, 41431 Slieta Nedjelja, Croatia
2lnstitute for Medical Research and Occupational Health, University of Zagreb, Ksaverska cesta 2, 41000 Zagreb, Croatia
It is hard to imagine any other type of animal product more tightly related to human health benefit than hyperimmune sera obtained on animals against some specific human infectious diseases or poisonings. The use of those preparations saved innumerable human lives and is alleviating the members of our species of pain for entire century. The therapy of certain, often fatal conditions in humans can not be successful without application of hyperimmune sera, despite the use of highly effective terapeutical drugs. Even more, according to our present knowledge the chances of exchange of hyperimmune animal sera in the future therapy of several human diseases with some industrially produced medicament are low. For animals actively participating in the process of sera production this enormus benefit humans is soul-stirringly painful. The reliance existing between owner and a horse, regardles of hard work to which is an animal sometimes forced to can not be achieved in the case of horses assigned to hyperimmune sera production. Virtually every contact with groomer means a new pain. The application of some antigens, especially snake poison are extremely painful to animal. The development of extensive tissue necrosis at the place of application is a rule. The animals are frequently bleeding and large amounts of blood are taken. Many of them develop amiloidosis almost inevitaby in this production. Even when they suffer of liver rupture humans do not let them die and they are cured recuperated and again used in production. Literary until the last drop of blood. The forehead mentioned sufferings can not be prevented or considerably improved even with the best will of groomers and veterinarians serving there. Certainly the number of ethical questions related to hyperimmune sera production will never be solved, but from moral point of humans they should be at least rezognized.
Stanična i tkivna mikrobijalna morfologija Cellular and tissue microbial morphology
Detection of human papilloma virus (HPV) genotypes in metastatic lesions of the invasive cervical cancer by in situ hybridization method
A. Pirkić, G. Grubišić, B. Rodé, Biserka Vukosavić-Cimić, Marija Matković
Department of Clinical Pathology “Prof. Dr. Ljudevit Jurak”,
and Clinic for Ginecology and Obstetrics, University Hospital “Sestre milosrdnice”, Zagreb, Croatia
In our previous study we have examined the various types of the CIN of the human uterine cervix as well as the invasive cervical cancer, to demonstrate the specific types of HPV DNA using three “cocktails” of biotynilated HPV DNA probes specific to HPV types 6/11, 16/18 and 31/33/51 (“Enzo Diagnostics”). In three invasive cancer, the types of HPV 16/18 and of HPV 31/33/51 were demonstrated.
In the here presented study the metastases of primary invasive cancer of cervix were examined for the demonstration of HPV genotypes in it. In the three of four invasive cancers, the involvement of adjacent lymphonodi by the metastatic spread from primary tumour was observed histologically. In the two cases of lymphonodular metastases the types of 31 /33/51 were detected. The hybridization signal in metastases was of a various intensity, and it was observed solely in the areas of a high grade of maturity of the metastatic tumours. The so called “pearls” and the adjacent tumour cells were the sites where the hybridization signal was mostly seen. The types of HPV DNA were detected in a single cells within the metastatic tissue. The demonstrated types of HPV DNA in the examined metastases as well as in the primary cervical cancer were the same. In the lymphonodi where metastatic process histologically was not seen, the hybridization signal was absent.
A contribution to pathogenic fungus determination in histologic sections
Institute for Medical Research and Occupational Health, University of Zagreb, Ksaverska cesta 2, 41000 Zagreb, Croatia
The increasing incidence of diseases in humans and domestic animals caused by pathogenic fungi is highly attributed to the widespread use of antibiotics as well as to a cytostatic and immunosuppresive therapy in human patients. Relatively high incidence of patients suffering of AIDS is favoring epidemiological condition as well. The diagnosis of fungal diseases is often difficult, and histopathological examination of biopsy or post mortem material is necessary for a reliable diagnosis. There are no essential differences among tissue response to fungi between humans and domestic mammals. Granulomatous or pyogranulomatous inflammation is the most common finding. Several special stains, in addition to routine HE stain of histo-sections are facilitating recognition of fungus morphology in the tissue. Most useful and widly used are PAS, Grocott’s and Grideley’s stain to fungy. Summarized, clue of characteristic morphology of most frequent pathogens follows. Aspergillus – septate, dychotomous branching hyphae, fruiting bodies. Blastomyces dermatidis – double contoured, single broad based bud. Candida- nonbranching pseudohyphae, spores. Coccidioides immites – large spheres with endospores. Cryptococcus neoformans – mucinous capsule, unequal budding. Histoplasma capsulatum – small organism, infrequently unequal budding, RE hyperplasia as tissue reaction. Mucor – non septate hyphae, irregular thickness, wide angle branching, prone to stain with HE. Some other microorganisms, such as alga. Prototheca or protozoarian Pneumocistis carinii, as well as polen particles might resemble fungi in the tissue, and lead to erroneous diagnosis.
Capsule structure among different genetic pools of Trichinella
A. Marinculić, Z. Kozarić, N. Božić, Palma E. Marinculić
Veterinary Faculty, University of Zagreb
Earlier investigations have employed a variety of methods for the study of the taxonomy of Trichinella. The most frequently studied biological characters are the reproductive capacity, the resistance of muscle larvae to low temperatures, host virulence and production of newborn larvae. Together these studies demonstrate that Trichinella is a genetically heterogeneous nematode group which was also defined by various isoenzymes structure analysis too. In the present report an analysis of 7 genetic pools of cystforming Trichinella is described for some morphological (nurse cell development and capsule structure) features. The results support the concept that the capsule structure can be a used as a good marker for determination of various species among the genus.
Cranial and cerebral echinococcosis and cysticercosis with special reference to our 4 cases
Jasna Talan-Hranilović, Smiljka Lambaša and L. Negovetić
Department of Clinical Pathology “Prof. Dr. Ljudevit Jurak”
University Hospital “Sestre milosrdnice”, Zagreb
Department of Neurosurgery, University Hospital “Sestre milosrdnice”, Zagreb
Cysticercosis and echinococcosis are the most frequent parasitic zoonosis of central nervous system (CNS) in humans in our country. Larva of parasit can be located and developed in nervous tissue of brain, arachnoidal membrane, ventricular cavities and spinal cord, but in bone of cranial vault and verthebra, as well. In the period of 1988-1922, 43 cases of these diseases with the symptoms of intracranial expansive process were diagnosed by neurosurgical byopsises. The surgery in all 4 cases revealed the cysts: 2 of them were histologically diagnosed as the hydatid cysts and the other 2 as the cysts of cysticercus cellulosae. Echinococcus cyst (hydatidosis) of a boy (10 y old) was found in the frontal brain lobe in the form of grate solitar cyst of typical histological structure with the scolex of the parasite. The case of echinococcus of a young woman (20 y old) in the last month of pregnancy, by whom the multiple cysts inside one gigantic cyst located intracranially, cranially and extracranially were found, was diagnosed. Surgery was performed after the delivery and in the extirpated cysts the multiple scolexes were found histologically. This case is interesting because of the possibility of intrauterinal child infection and congenital echinococcossis, which is extremly rare.
Cysticercosis was diagnosed in the man (73 y old) with cyst of arachnoidal membrane of brain base at the clivus and in the man (50 y old) with cysticercous cyst in the fourth ventricle and on the medulla oblongata. Only cyst membrane was given for the pathohistological analysis. The analised cysts were of typical structure and in the ventricular cyst scolex of the parasit was found.
The 4 cases of echinococcosis and cysticercosis are the only cases of the parasitic diseases of CNS, diagnosed in about 4000 of neurosurgical byopsises.
Nove metode u patologiji
New methods in pathological examination
Staining and visualisation of the argyrophilic proteins of the nucleolar organiser region in paraffin sections (AgNOR)
Smiljka Lambaša, J. Matić
Clinical Hospital “Sestre milosrdnice”, Department of Clinical Pathology “Prof. Dr. Ljudevit Jurak”, Zagreb, Croatia
AgNOR areas in interphase nuclei of normal and neoplastic cells correlate with the proliferative activity. Therefor the AgNOR enumeration is of prognostic value both in malignant and displastic lesions.
The aim of our study was to visualise AgNOR dots clarely and selectively trying to avoid possible artifacts.
AgNOR staining was carried out using a solution of one volume of 2% gelatine in 1 % aqueous formic acid and two volumes of 50% silver nitrate. The duration of impregnation was between 10 and 30 minutes with or without contrastain.
The aplication of the AgNOR method gives structures which are black; other structures often color brown to yellow. We shall present our results and point the possible errors in staining procedure.
Intercellular adhesion molecule in the immune response
Department of Clinical Pathology University Hospital “Sestre milosrdnice”, Zagreb
Cell-Cell adherence is a mandatory step in strengthening the interactions between lympocites and target cells during antigen presentation and effector cell functions in the immune response. Various adhesion molecules are involved in this antigen-independet adhesion process. The intercellular adhesion molecule -1 (ICAM1, CD 54), a glycoprotein which belongs to the imunoglobulin superfamily of cell surface receptors, is the natural ligand for the lymphocyte – associated antigen – 1 (LFA-1, CDlla/CD18).
Binding of LFA-1 on lymphocytes to ICAM-1 on target cells precedes, or cocurrently occurs with recognition ofantigen. ICAM-1 is an inducible molecule, since various cytokines induce both IN VIVO and IN VITRO its de novo expression on variety of normal human cell types (keratinocytes, melanocytes, endothelial cells, fibroblasts). The discovery of adhesion molecules opens new prospects for interventions in immune phenomena.
The spectrum of possibilities ranges from immunostimulation to immunosuppresion directed towards individual molecules and restricted to defined coompartmens.
The Strategies for immune intervention at the level of adhesion structures have already advanced beyond the stage of mere hypothesis, and in certain conditions such as graft rejection they have progressed to techniques which will soon be ready for clinical use.
Kontrastiranje ekstracelularnih elemenata s taninskom kiselinom
Đ. Vranešić i Karmela Šuvak
Nova bolnica Zagreb, Služba za patologiju
Standardna metoda kontrastiranja ultratankih prereza ima najbolji efekt na stanice i stanične organele, dok se ekstracelulami elementi prikazuju slabije, nejasno ili čak ostaju nekontrastirani, pa takva mjesta imponiraju kao prazni prostori. To se osobito odnosi na kolagen, elastin i proteoglikane, koji se vrlo teško i slabo kontrastiraju sa uranil acetatom i olovnim citratom.
Prikazali smo metodu kojom se postiže povećanje kontrasta ekstracelularnih tkivnih komponenti. Metoda se sastoji od predkontrastiranja mrežica u otopini taninske kiseline. Metoda je brza i jednostavna, a funkcionira u svim slučajevima, bez obzira na prethodnu preparaciju uzorka. Dodatna korist od primjene ove metode je znatno bolje kontrastiranje miofibrila i glikogena.
Testing of a new polyclonal antibody against BPC 157 peptide
Ljubica Škreblin, S. Seiwerth, B. Turković, M. Mildner
Institute of Pathology, Medical Faculty University of Zagreb
A new gastricjuice peptide, code named BPC, was recently isolated and partially aminosequenced. A synthetic pentadekapeptide, correlating with,the working fragment of the peptide, was produced and named BPC 157. Polyclonal antybodies were produced in rabitts, purrified by immunoadsorbtion and tested by immunelectrophoresis and RIA. Immunohistochemically the whole serum and the purrified antibody were tested under different conditions and on different tissues. Here we are presenting the immunohistochemical testing for the newly produced antibody.
Resorcin-fuchsin-van Gieson, a method for simultaneous demonstration of connective tissue and elastic fibres
Jadranka Kajganić, S. Seiwerth
Institute of Pathology, Medical Faculty University of Zagreb
In morphological analysis special stains are often used to mark tissue components in order to help us to make up pathological changes or in their differential diagnosis. For demonstration of elastic fibres in our routine work we use orcein while connective tissue is demonstrated by van Gieson or Mallory trichrome. In stead of orcein resorcin-fuchsin can be used (after Weigert). The dye can be aquired commercially. Simultaneous identification of elastic and connective tissue is made possible by combining Weigerts and van Gieson methods. By using resorcin fuchsin instead of orcein we significantly reduce the performing time. The method is usefull in routine diagnosis, particularly in cardio-vascular pathology. We present the method with chemicals made in our own laboratory.
Patohistological techniques training – 5 years of experience
Nevenka Markovinović, Spomenka Manojlović
Institute of Pathology, Medical faculty University of Zagreb, Zagreb, Croatia
Five years ago a practical course in histology techniques was introduced in the curriculum for students of the School for medical laboratory engineers. The aim of this course is to establish a basic practical knowledge and skill in elements of histological techniques and to prepare the student for a rapid incorporation in the operative procedures of a standard histology laboratory. In this a practical training in steps preceding the staining (such as fixation, processing, embedding, cutting) is performed supported by abroad theoretical background. Also some special techniques used commonly in the routine pathohistology laboratory are performed by each student. More sophisticated techniques, such as immunohistochemistry and electron microscopy are only demonstrated. A serious limitation in our efforts is the tight schedule and our goal is to include also two practical weeks in the curriculum. So the students could be much better incorporated in the daily routine and get the opportunity to apply their knowledge.
Undecalcified bone histology in pathological examination
Vesna Kušec, Ana Marušić, Marija Šaban and Đurdica Car
Department for Clinical Laboratory Diagnosis, Clinical Haspital Center Zagreb and Zagreb University, School of Medicine; and Department of Anatomy, Zagreb University School of Medicine, Zagreb, Croatia
Diagnosis of alteration of bone metabolism is an important factor in evaluating the prognosis and therapy in many diseases which affect skeletal system. The best method for evaluating bone metabolism is morphological examination of bone biopsies embedded in methylmetacrylate. Bone specimen obtained after biopsy and measuring at least 5 mm in diameter is fixed in 70% ethanol, dehydrated in increasing concentrations of ethanol and embedded in methylmetacrylate. The polymerization of the plastic resin takes place at 50-60°C. The resulting plastic block is cut into 3 mthin sections using a microtome knife for hard tissue. Sections can be stained using different staining protocols (toluidine blue, Goldner’s trichrome stain, Ladewik stain) which allow differentiation of various bone compartments (mineralized bone, osteoid, bone cells) and their alterations. Modification of the technique involving addition of glycolmetacrylate and low polymerization temperature allows susequent immunohistochemistry of undecalcified bone sections.
Alterations of bone metabolism can be evaluated by computerized histomorphometry (available at the Department for Clinical Diagnosis). Static (e.g. trabecular bone volume, osteoid thickness, resorptive and formative surfaces, number of osteoblasts and osteoclasts) and dynamic parameters (bone apposition rate, using tetracycline labels) can be measured. Such evaluation is of great value in diagnostics of bone pathology.
Histochemical staining for DNA and RNA with Methylgreen-pyronin
Đurđa Jurin, Sanja Đačić and Keti Mišetić
Insfitute of Pathology, Medical faculty University of Zagreb
In nucleic proteins constituents histochemical methods for demonstration of nucleic acids (DNA and RNA) are available. The most popular procedures for nucleic acid demonstration are Feulgens reaction for DNA and the combined methyl green – pyronin reaction for demonstration of both DNA and RNA. This reaction represents a relativelly easy to perform tool in differential diagnosis of haemathological disorders. The accumulation of pyronin-positive RNA in benign and malignant plasma cells enables us to identifie them under various conditions.
Immunofluorescence in kidney biopsy analysis
Z. Batik, Sanja Čužić
Institute for Pathology, KBC Rebro, Zagreb, Croatia
Immunofluorescence is a critical method in a assessment of renal biopsy specimen in order to demonstrate the immunoglobulins, complement components and fibrin which can be part of pathogenic immunocomplexes.
Detailed immunofluorescence method on frozen aceton fixed renal biopsy sections with FITC (fluorescein-iso-thicyanide) labeled antibodies, as used in our laboratory, will be presented.
Immunohistochemical methods on frozen sections and fixed material
M. Rogina, Mira Šćukanec-Špoljar
Institute for Pathology, KBC Rebro, Zagreb, Croatia
In our faboratory peroxidase – anti – peroxidase (PAP) immunohistochemical method is used to detect various tissue antigens. We focus on interstitium inflammatory cells found in glomerulonephritis.
The assessment to delicate renal biopsy specimen regarding fixation, antigen preservatin and staining will be discussed.